Parkinson disease-associated mutations in LRRK2 cause centrosomal defects via Rab8a phosphorylation.

PubMed

Madero-Pérez, Jesús; Fdez, Elena; Fernández, Belén; Lara Ordóñez, Antonio J; Blanca Ramírez, Marian; Gómez-Suaga, Patricia; Waschbüsch, Dieter; Lobbestael, Evy; Baekelandt, Veerle; Nairn, Angus C; Ruiz-Martínez, Javier; Aiastui, Ana; López de Munain, Adolfo; Lis, Pawel; Comptdaer, Thomas; Taymans, Jean-Marc; Chartier-Harlin, Marie-Christine; Beilina, Alexandria; Gonnelli, Adriano; Cookson, Mark R; Greggio, Elisa; Hilfiker, Sabine

2018-01-23

Mutations in LRRK2 are a common genetic cause of Parkinson's disease (PD). LRRK2 interacts with and phosphorylates a subset of Rab proteins including Rab8a, a protein which has been implicated in various centrosome-related events. However, the cellular consequences of such phosphorylation remain elusive. Human neuroblastoma SH-SY5Y cells stably expressing wildtype or pathogenic LRRK2 were used to test for polarity defects in the context of centrosomal positioning. Centrosomal cohesion deficits were analyzed from transiently transfected HEK293T cells, as well as from two distinct peripheral cell types derived from LRRK2-PD patients. Kinase assays, coimmunoprecipitation and GTP binding/retention assays were used to address Rab8a phosphorylation by LRRK2 and its effects in vitro. Transient transfections and siRNA experiments were performed to probe for the implication of Rab8a and its phosphorylated form in the centrosomal deficits caused by pathogenic LRRK2. Here, we show that pathogenic LRRK2 causes deficits in centrosomal positioning with effects on neurite outgrowth, cell polarization and directed migration. Pathogenic LRRK2 also causes deficits in centrosome cohesion which can be detected in peripheral cells derived from LRRK2-PD patients as compared to healthy controls, and which are reversed upon LRRK2 kinase inhibition. The centrosomal cohesion and polarity deficits can be mimicked when co-expressing wildtype LRRK2 with wildtype but not phospho-deficient Rab8a. The centrosomal defects induced by pathogenic LRRK2 are associated with a kinase activity-dependent increase in the centrosomal localization of phosphorylated Rab8a, and are prominently reduced upon RNAi of Rab8a. Our findings reveal a new function of LRRK2 mediated by Rab8a phosphorylation and related to various centrosomal defects.

The Tumor Suppressor Cdkn3 Is Required for Maintaining the Proper Number of Centrosomes by Regulating the Centrosomal Stability of Mps1.

PubMed

Srinivas, Vinayaka; Kitagawa, Mayumi; Wong, Jasmine; Liao, Pei-Ju; Lee, Sang Hyun

2015-11-24

Supernumerary centrosomes promote the assembly of abnormal spindles in many human cancers. The observation that modest changes in the centrosomal levels of Mps1 kinase can cause centrosome overduplication in human cells suggests the existence of a regulatory system that may tightly control its centrosomal stability. Here, we show that Cdkn3, a Cdk-associated phosphatase, prevents Mps1-mediated centrosome overduplication. We identify Cdkn3 as a direct binding partner of Mps1. The interaction between Mps1 and Cdkn3 is required for Mps1 to recruit Cdkn3 to centrosomes. Subsequently, Mps1-bound Cdkn3 forms a regulatory system that controls the centrosomal levels of Mps1 through proteasome-mediated degradation and thereby prevents Mps1-mediated centrosome overduplication. Conversely, knockdown of Cdkn3 stabilizes Mps1 at centrosomes, which promotes centrosome overduplication. We suggest that Mps1 and Cdkn3 form a self-regulated feedback loop at centrosomes to tightly control the centrosomal levels of Mps1, which prevents centrosome overduplication in human cells. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Antizyme restrains centrosome amplification by regulating the accumulation of Mps1 at centrosomes.

PubMed

Kasbek, Christopher; Yang, Ching-Hui; Fisk, Harold A

2010-11-15

Extra centrosomes are found in many tumors, and their appearance is an early event that can generate aberrant mitotic spindles and aneuploidy. Because the failure to appropriately degrade the Mps1 protein kinase correlates with centrosome overproduction in tumor-derived cells, defects in the factors that promote Mps1 degradation may contribute to extra centrosomes in tumors. However, while we have recently characterized an Mps1 degradation signal, the factors that regulate Mps1 centrosomal Mps1 are unknown. Antizyme (OAZ), a mediator of ubiquitin-independent degradation and a suspected tumor suppressor, was recently shown to localize to centrosomes and modulate centrosome overproduction, but the known OAZ substrates were not responsible for its effect on centrosomes. We have found that OAZ exerts its effect on centrosomes via Mps1. OAZ promotes the removal of Mps1 from centrosomes, and centrosome overproduction caused by reducing OAZ activity requires Mps1. OAZ binds to Mps1 via the Mps1 degradation signal and modulates the function of Mps1 in centrosome overproduction. Moreover, OAZ regulates the canonical centrosome duplication cycle, and reveals a function for Mps1 in procentriole assembly. Together, our data suggest that OAZ restrains the assembly of centrioles by controlling the levels of centrosomal Mps1 through the Cdk2-regulated Mps1 degradation signal.

Interphase centrosome organization by the PLP-Cnn scaffold is required for centrosome function

PubMed Central

Lerit, Dorothy A.; Jordan, Holly A.; Poulton, John S.; Fagerstrom, Carey J.; Galletta, Brian J.; Peifer, Mark

2015-01-01

Pericentriolar material (PCM) mediates the microtubule (MT) nucleation and anchoring activity of centrosomes. A scaffold organized by Centrosomin (Cnn) serves to ensure proper PCM architecture and functional changes in centrosome activity with each cell cycle. Here, we investigate the mechanisms that spatially restrict and temporally coordinate centrosome scaffold formation. Focusing on the mitotic-to-interphase transition in Drosophila melanogaster embryos, we show that the elaboration of the interphase Cnn scaffold defines a major structural rearrangement of the centrosome. We identify an unprecedented role for Pericentrin-like protein (PLP), which localizes to the tips of extended Cnn flares, to maintain robust interphase centrosome activity and promote the formation of interphase MT asters required for normal nuclear spacing, centrosome segregation, and compartmentalization of the syncytial embryo. Our data reveal that Cnn and PLP directly interact at two defined sites to coordinate the cell cycle–dependent rearrangement and scaffolding activity of the centrosome to permit normal centrosome organization, cell division, and embryonic viability. PMID:26150390

Interphase centrosome organization by the PLP-Cnn scaffold is required for centrosome function.

PubMed

Lerit, Dorothy A; Jordan, Holly A; Poulton, John S; Fagerstrom, Carey J; Galletta, Brian J; Peifer, Mark; Rusan, Nasser M

2015-07-06

Pericentriolar material (PCM) mediates the microtubule (MT) nucleation and anchoring activity of centrosomes. A scaffold organized by Centrosomin (Cnn) serves to ensure proper PCM architecture and functional changes in centrosome activity with each cell cycle. Here, we investigate the mechanisms that spatially restrict and temporally coordinate centrosome scaffold formation. Focusing on the mitotic-to-interphase transition in Drosophila melanogaster embryos, we show that the elaboration of the interphase Cnn scaffold defines a major structural rearrangement of the centrosome. We identify an unprecedented role for Pericentrin-like protein (PLP), which localizes to the tips of extended Cnn flares, to maintain robust interphase centrosome activity and promote the formation of interphase MT asters required for normal nuclear spacing, centrosome segregation, and compartmentalization of the syncytial embryo. Our data reveal that Cnn and PLP directly interact at two defined sites to coordinate the cell cycle-dependent rearrangement and scaffolding activity of the centrosome to permit normal centrosome organization, cell division, and embryonic viability.

Phosphorylation of the centrosomal protein, Cep169, by Cdk1 promotes its dissociation from centrosomes in mitosis.

PubMed

Mori, Yusuke; Inoue, Yoko; Taniyama, Yuki; Tanaka, Sayori; Terada, Yasuhiko

2015-12-25

Cep169 is a centrosomal protein conserved among vertebrates. In our previous reports, we showed that mammalian Cep169 interacts and collaborates with CDK5RAP2 to regulate microtubule (MT) dynamics and stabilization. Although Cep169 is required for MT regulation, its precise cellular function remains largely elusive. Here we show that Cep169 associates with centrosomes during interphase, but dissociates from these structures from the onset of mitosis, although CDK5RAP2 (Cep215) is continuously located at the centrosomes throughout cell cycle. Interestingly, treatment with purvalanol A, a Cdk1 inhibitor, nearly completely blocked the dissociation of Cep169 from centrosomes during mitosis. In addition, mass spectrometry analyses identified 7 phosphorylated residues of Cep169 corresponding to consensus phosphorylation sequence for Cdk1. These data suggest that the dissociation of Cep169 from centrosomes is controlled by Cdk1/Cyclin B during mitosis, and that Cep169 might regulate MT dynamics of mitotic spindle. Copyright © 2015 Elsevier Inc. All rights reserved.

Sequential activities of Dynein, Mud and Asp in centrosome-spindle coupling maintain centrosome number upon mitosis.

PubMed

Bosveld, Floris; Ainslie, Anna; Bellaïche, Yohanns

2017-10-15

Centrosomes nucleate microtubules and are tightly coupled to the bipolar spindle to ensure genome integrity, cell division orientation and centrosome segregation. While the mechanisms of centrosome-dependent microtubule nucleation and bipolar spindle assembly have been the focus of numerous works, less is known about the mechanisms ensuring the centrosome-spindle coupling. The conserved NuMA protein (Mud in Drosophila ) is best known for its role in spindle orientation. Here, we analyzed the role of Mud and two of its interactors, Asp and Dynein, in the regulation of centrosome numbers in Drosophila epithelial cells. We found that Dynein and Mud mainly initiate centrosome-spindle coupling prior to nuclear envelope breakdown (NEB) by promoting correct centrosome positioning or separation, while Asp acts largely independently of Dynein and Mud to maintain centrosome-spindle coupling. Failure in the centrosome-spindle coupling leads to mis-segregation of the two centrosomes into one daughter cell, resulting in cells with supernumerary centrosomes during subsequent divisions. Altogether, we propose that Dynein, Mud and Asp operate sequentially during the cell cycle to ensure efficient centrosome-spindle coupling in mitosis, thereby preventing centrosome mis-segregation to maintain centrosome number. © 2017. Published by The Company of Biologists Ltd.

SUMO and Nucleocytoplasmic Transport.

PubMed

Ptak, Christopher; Wozniak, Richard W

2017-01-01

The transport of proteins between the nucleus and cytoplasm occurs through nuclear pore complexes and is facilitated by numerous transport factors. These transport processes are often regulated by post-translational modification or, reciprocally, transport can function to control post-translational modifications through regulated transport of key modifying enzymes. This interplay extends to relationships between nucleocytoplasmic transport and SUMO-dependent pathways. Examples of protein sumoylation inhibiting or stimulating nucleocytoplasmic transport have been documented, both through its effects on the physical properties of cargo molecules and by directly regulating the functions of components of the nuclear transport machinery. Conversely, the nuclear transport machinery regulates the localization of target proteins and enzymes controlling dynamics of sumoylation and desumoylation thereby affecting the sumoylation state of target proteins. These inter-relationships between SUMO and the nucleocytoplasmic transport machinery, and the varied ways in which they occur, are discussed.

Activation of maternal centrosomes in unfertilized sea urchin eggs

NASA Technical Reports Server (NTRS)

Schatten, H.; Walter, M.; Biessmann, H.; Schatten, G.

1992-01-01

Centrosomes are undetectable in unfertilized sea urchin eggs, and normally the sperm introduces the cell's microtubule-organizing center (MTOC) at fertilization. However, artificial activation or parthenogenesis triggers microtubule assembly in the unfertilized egg, and this study explores the reappearance and behavior of the maternal centrosome. During activation with A23187 or ammonia, microtubules appear first at the cortex; centrosomal antigen is detected diffusely throughout the entire cytoplasm. Later, the centrosome becomes more distinct and organizes a radial microtubule shell, and eventually a compact centrosome at the egg center organizes a monaster. In these activated eggs, centrosomes undergo cycles of compaction and decompaction in synchrony with the chromatin, which also undergoes cycles of condensation and decondensation. Parthenogenetic activation with heavy water (50% D2O) or the microtubule-stabilizing drug taxol (10 microM) induces numerous centrosomal foci in the unfertilized sea urchin egg. Within 15 min after incubation in D2O, numerous fine centrosomal foci are detected, and they organize a connected network of numerous asters which fill the entire egg. Taxol induces over 100 centrosomal foci by 15 min after treatment, which organize a corresponding number of asters. The centrosomal material in either D2O- or taxol-treated eggs aggregates with time to form fewer but denser foci, resulting in fewer and larger asters. Fertilization of eggs pretreated with either D2O or taxol shows that the paternal centrosome is dominant over the maternal centrosome. The centrosomal material gradually becomes associated with the enlarged sperm aster. These experiments demonstrate that maternal centrosomal material is present in the unfertilized egg, likely as dispersed undetectable material, which can be activated without paternal contributions. At fertilization, paternal centrosomes become dominant over the maternal centrosomal material.

The G1 phase Cdks regulate the centrosome cycle and mediate oncogene-dependent centrosome amplification

PubMed Central

2011-01-01

Because centrosome amplification generates aneuploidy and since centrosome amplification is ubiquitous in human tumors, a strong case is made for centrosome amplification being a major force in tumor biogenesis. Various evidence showing that oncogenes and altered tumor suppressors lead to centrosome amplification and aneuploidy suggests that oncogenes and altered tumor suppressors are a major source of genomic instability in tumors, and that they generate those abnormal processes to initiate and sustain tumorigenesis. We discuss how altered tumor suppressors and oncogenes utilize the cell cycle regulatory machinery to signal centrosome amplification and aneuploidy. PMID:21272329

Chronic centrosome amplification without tumorigenesis

PubMed Central

Vitre, Benjamin; Holland, Andrew J.; Kulukian, Anita; Shoshani, Ofer; Hirai, Maretoshi; Wang, Yin; Maldonado, Marcus; Cho, Thomas; Boubaker, Jihane; Swing, Deborah A.; Tessarollo, Lino; Evans, Sylvia M.; Fuchs, Elaine; Cleveland, Don W.

2015-01-01

Centrosomes are microtubule-organizing centers that facilitate bipolar mitotic spindle assembly and chromosome segregation. Recognizing that centrosome amplification is a common feature of aneuploid cancer cells, we tested whether supernumerary centrosomes are sufficient to drive tumor development. To do this, we constructed and analyzed mice in which centrosome amplification can be induced by a Cre-recombinase–mediated increase in expression of Polo-like kinase 4 (Plk4). Elevated Plk4 in mouse fibroblasts produced supernumerary centrosomes and enhanced the expected mitotic errors, but proliferation continued only after inactivation of the p53 tumor suppressor. Increasing Plk4 levels in mice with functional p53 produced centrosome amplification in liver and skin, but this did not promote spontaneous tumor development in these tissues or enhance the growth of chemically induced skin tumors. In the absence of p53, Plk4 overexpression generated widespread centrosome amplification, but did not drive additional tumors or affect development of the fatal thymic lymphomas that arise in animals lacking p53. We conclude that, independent of p53 status, supernumerary centrosomes are not sufficient to drive tumor formation. PMID:26578792

14-3-3γ Prevents Centrosome Amplification and Neoplastic Progression.

PubMed

Mukhopadhyay, Amitabha; Sehgal, Lalit; Bose, Arunabha; Gulvady, Anushree; Senapati, Parijat; Thorat, Rahul; Basu, Srikanta; Bhatt, Khyati; Hosing, Amol S; Balyan, Renu; Borde, Lalit; Kundu, Tapas K; Dalal, Sorab N

2016-06-02

More than 80% of malignant tumors show centrosome amplification and clustering. Centrosome amplification results from aberrations in the centrosome duplication cycle, which is strictly coordinated with DNA-replication-cycle. However, the relationship between cell-cycle regulators and centrosome duplicating factors is not well understood. This report demonstrates that 14-3-3γ localizes to the centrosome and 14-3-3γ loss leads to centrosome amplification. Loss of 14-3-3γ results in the phosphorylation of NPM1 at Thr-199, causing early centriole disjunction and centrosome hyper-duplication. The centrosome amplification led to aneuploidy and increased tumor formation in mice. Importantly, an increase in passage of the 14-3-3γ-knockdown cells led to an increase in the number of cells containing clustered centrosomes leading to the generation of pseudo-bipolar spindles. The increase in pseudo-bipolar spindles was reversed and an increase in the number of multi-polar spindles was observed upon expression of a constitutively active 14-3-3-binding-defective-mutant of cdc25C (S216A) in the 14-3-3γ knockdown cells. The increase in multi-polar spindle formation was associated with decreased cell viability and a decrease in tumor growth. Our findings uncover the molecular basis of regulation of centrosome duplication by 14-3-3γ and inhibition of tumor growth by premature activation of the mitotic program and the disruption of centrosome clustering.

Co-translational protein targeting facilitates centrosomal recruitment of PCNT during centrosome maturation in vertebrates

PubMed Central

Mahe, Karan; Ou, Tingyoung; Castro, Noemi M; Christensen, Lana N; Cheung, Lee; Jiang, Xueer; Yoon, Daniel; Huang, Bo

2018-01-01

As microtubule-organizing centers of animal cells, centrosomes guide the formation of the bipolar spindle that segregates chromosomes during mitosis. At mitosis onset, centrosomes maximize microtubule-organizing activity by rapidly expanding the pericentriolar material (PCM). This process is in part driven by the large PCM protein pericentrin (PCNT), as its level increases at the PCM and helps recruit additional PCM components. However, the mechanism underlying the timely centrosomal enrichment of PCNT remains unclear. Here, we show that PCNT is delivered co-translationally to centrosomes during early mitosis by cytoplasmic dynein, as evidenced by centrosomal enrichment of PCNT mRNA, its translation near centrosomes, and requirement of intact polysomes for PCNT mRNA localization. Additionally, the microtubule minus-end regulator, ASPM, is also targeted co-translationally to mitotic spindle poles. Together, these findings suggest that co-translational targeting of cytoplasmic proteins to specific subcellular destinations may be a generalized protein targeting mechanism. PMID:29708497

A novel mechanism of E2F1 regulation via nucleocytoplasmic shuttling: determinants of nuclear import and export.

PubMed

Ivanova, Iordanka A; Vespa, Alisa; Dagnino, Lina

2007-09-01

E2F1 is a transcription factor central for cell survival, proliferation, and repair following genomic insult. Depending on the cell type and conditions, E2F1 can induce apoptosis in transformed cells, behaving as a tumour suppressor, or impart growth advantages favouring tumour formation. The pleiotropic functions of E2F1 are a likely consequence of its ability to transcriptionally control a wide variety of target genes, and require tight regulation of its activity at multiple levels. Although sequestration of proteins to particular cellular compartments is a well-established regulatory mechanism, virtually nothing is known about its contribution to modulation of E2F1 target gene expression. We have examined the subcellular trafficking of E2F1 and, contrary to the widely held notion that this factor is constitutively nuclear, we now demonstrate that it is subjected to continuous nucleocytoplasmic shuttling. We have also defined two nuclear localization domains and a nuclear export region, which mediates CRM1-dependent transit out of the nucleus. The predominant subcellular location of E2F1 is likely determined by the balance between the activity of nuclear import and export domains, and can be modulated by differentiation stimuli in epidermal cells. Thus, we have identified a hitherto unrecognized mechanism to control E2F1 function through modulation of its subcellular localization.

Nucleo-cytoplasmic shuttling dynamics of the transcriptional regulators XYR1 and CRE1 under conditions of cellulase and xylanase gene expression in Trichoderma reesei

PubMed Central

Lichius, Alexander; Seidl-Seiboth, Verena; Seiboth, Bernhard; Kubicek, Christian P

2014-01-01

Trichoderma reesei is a model for investigating the regulation of (hemi-)cellulase gene expression. Cellulases are formed adaptively, and the transcriptional activator XYR1 and the carbon catabolite repressor CRE1 are main regulators of their expression. We quantified the nucleo-cytoplasmic shuttling dynamics of GFP-fusion proteins of both transcription factors under cellulase and xylanase inducing conditions, and correlated their nuclear presence/absence with transcriptional changes. We also compared their subcellular localization in conidial germlings and mature hyphae. We show that cellulase gene expression requires de novo biosynthesis of XYR1 and its simultaneous nuclear import, whereas carbon catabolite repression is regulated through preformed CRE1 imported from the cytoplasmic pool. Termination of induction immediately stopped cellulase gene transcription and was accompanied by rapid nuclear degradation of XYR1. In contrast, nuclear CRE1 rapidly decreased upon glucose depletion, and became recycled into the cytoplasm. In mature hyphae, nuclei containing activated XYR1 were concentrated in the colony center, indicating that this is the main region of XYR1 synthesis and cellulase transcription. CRE1 was found to be evenly distributed throughout the entire mycelium. Taken together, our data revealed novel aspects of the dynamic shuttling and spatial bias of the major regulator of (hemi-)cellulase gene expression, XYR1, in T. reesei. PMID:25302561

The Effects of Modeled Microgravity on Nucleocytoplasmic Localization of Human Apurinic/Apyrimidinic

NASA Technical Reports Server (NTRS)

Gonda, Steve; Jackson, E.B.

2004-01-01

Exposure to space radiation and microgravity occurs to humans during space flight. In order to have accurate risk estimations, answering questions to whether increased DNA damage seen during space flight in modified by microgravity are important. Several studies have examined whether intercellular repair of radiation-induced DNA lesions are modified by microgravity. Results from these studies show no modification of the repair processes due to microgravity. However, it is known that in studies not involving radiation that microgravity interferes with normal development. Interestingly, there is no data that attempts to analyze the possible effects of microgravity on the trafficking of DNA repair proteins. In this study, we analyze the effects of modeled microgravity on nucleocytoplasmic shuttling of the human DNA repair enzyme apurinic/apyrimidinic endonuclease 1 (APE1/Ref1) which is involved in base excision repair. We examined nuclear translocation of APE1 using enhanced green fluorescent protein (EGFP) fused to APE1 as a reporter. While APE1 under normal gravity showed normal nuclear localization, APE1 nuclear localization under modeled microgravity was decreased. These results suggest that nucleocytoplasmic translocation of APE1 is modified under modeled microgravity.

Centrosome movement in the early divisions of Caenorhabditis elegans: A cortical site determining centrosome position

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hyman, A.A.

1989-09-01

In Caenorhabditis elegans embryos, early blastomeres of the P cell lineage divide successively on the same axis. This axis is a consequence of the specific rotational movement of the pair of centrosomes and nucleus. A laser has been used to perturb the centrosome movements that determine the pattern of early embryonic divisions. The results support a previously proposed model in which a centrosome rotates towards its correct position by shortening of connections, possibly microtubules, between a centrosome and a defined site on the cortex of the embryo.

The Centrosome-Specific Phosphorylation of Cnn by Polo/Plk1 Drives Cnn Scaffold Assembly and Centrosome Maturation

PubMed Central

Conduit, Paul T.; Feng, Zhe; Richens, Jennifer H.; Baumbach, Janina; Wainman, Alan; Bakshi, Suruchi D.; Dobbelaere, Jeroen; Johnson, Steven; Lea, Susan M.; Raff, Jordan W.

2014-01-01

Summary Centrosomes are important cell organizers. They consist of a pair of centrioles surrounded by pericentriolar material (PCM) that expands dramatically during mitosis—a process termed centrosome maturation. How centrosomes mature remains mysterious. Here, we identify a domain in Drosophila Cnn that appears to be phosphorylated by Polo/Plk1 specifically at centrosomes during mitosis. The phosphorylation promotes the assembly of a Cnn scaffold around the centrioles that is in constant flux, with Cnn molecules recruited continuously around the centrioles as the scaffold spreads slowly outward. Mutations that block Cnn phosphorylation strongly inhibit scaffold assembly and centrosome maturation, whereas phosphomimicking mutations allow Cnn to multimerize in vitro and to spontaneously form cytoplasmic scaffolds in vivo that organize microtubules independently of centrosomes. We conclude that Polo/Plk1 initiates the phosphorylation-dependent assembly of a Cnn scaffold around centrioles that is essential for efficient centrosome maturation in flies. PMID:24656740

The centrosome-specific phosphorylation of Cnn by Polo/Plk1 drives Cnn scaffold assembly and centrosome maturation.

PubMed

Conduit, Paul T; Feng, Zhe; Richens, Jennifer H; Baumbach, Janina; Wainman, Alan; Bakshi, Suruchi D; Dobbelaere, Jeroen; Johnson, Steven; Lea, Susan M; Raff, Jordan W

2014-03-31

Centrosomes are important cell organizers. They consist of a pair of centrioles surrounded by pericentriolar material (PCM) that expands dramatically during mitosis-a process termed centrosome maturation. How centrosomes mature remains mysterious. Here, we identify a domain in Drosophila Cnn that appears to be phosphorylated by Polo/Plk1 specifically at centrosomes during mitosis. The phosphorylation promotes the assembly of a Cnn scaffold around the centrioles that is in constant flux, with Cnn molecules recruited continuously around the centrioles as the scaffold spreads slowly outward. Mutations that block Cnn phosphorylation strongly inhibit scaffold assembly and centrosome maturation, whereas phosphomimicking mutations allow Cnn to multimerize in vitro and to spontaneously form cytoplasmic scaffolds in vivo that organize microtubules independently of centrosomes. We conclude that Polo/Plk1 initiates the phosphorylation-dependent assembly of a Cnn scaffold around centrioles that is essential for efficient centrosome maturation in flies. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

A fraction of Crm1 locates at centrosomes by its CRIME domain and regulates the centrosomal localization of pericentrin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Qinying; Jiang, Qing; Zhang, Chuanmao, E-mail: zhangcm@pku.edu.cn

Crm1 plays a role in exporting proteins containing nuclear export signals (NESs) from the nucleus to the cytoplasm. Some proteins that are capable of interacting with Ran/Crm1 were reported to be localized at centrosomes and to function as centrosome checkpoints. But it remains unclear how Crm1 locates at centrosomes. In this study, we found that a fraction of Crm1 is located at centrosomes through its N-terminal CRM1, importin {beta} etc. (CRIME) domain, which is responsible for interacting with RanGTP, suggesting that Crm1 might target to centrosomes through binding centrosomal RanGTP. Moreover, overexpression of the CRIME domain, which is free ofmore » NES binding domain, resulted in the dissociation of pericentrin and {gamma}-tubulin complex from centrosomes and the disruption of microtubule nucleation. Deficiency of Crm1 provoked by RNAi also decreased the spindle poles localization of pericentrin and {gamma}-tubulin complex, coupled with mitotic defects. Since pericentrin was sensitive to Crm1 specific inhibitor leptomycin B, we propose that the centrosomal Crm1 might interact with pericentrin and regulate the localization and function of pericentrin at centrosomes.« less

Structural centrosome aberrations promote non-cell-autonomous invasiveness.

PubMed

Ganier, Olivier; Schnerch, Dominik; Oertle, Philipp; Lim, Roderick Yh; Plodinec, Marija; Nigg, Erich A

2018-05-02

Centrosomes are the main microtubule-organizing centers of animal cells. Although centrosome aberrations are common in tumors, their consequences remain subject to debate. Here, we studied the impact of structural centrosome aberrations, induced by deregulated expression of ninein-like protein (NLP), on epithelial spheres grown in Matrigel matrices. We demonstrate that NLP-induced structural centrosome aberrations trigger the escape ("budding") of living cells from epithelia. Remarkably, all cells disseminating into the matrix were undergoing mitosis. This invasive behavior reflects a novel mechanism that depends on the acquisition of two distinct properties. First, NLP-induced centrosome aberrations trigger a re-organization of the cytoskeleton, which stabilizes microtubules and weakens E-cadherin junctions during mitosis. Second, atomic force microscopy reveals that cells harboring these centrosome aberrations display increased stiffness. As a consequence, mitotic cells are pushed out of mosaic epithelia, particularly if they lack centrosome aberrations. We conclude that centrosome aberrations can trigger cell dissemination through a novel, non-cell-autonomous mechanism, raising the prospect that centrosome aberrations contribute to the dissemination of metastatic cells harboring normal centrosomes. © 2018 The Authors. Published under the terms of the CC BY NC ND 4.0 license.

Fanconi anemia complementation group A (FANCA) localizes to centrosomes and functions in the maintenance of centrosome integrity.

PubMed

Kim, Sunshin; Hwang, Soo Kyung; Lee, Mihee; Kwak, Heejin; Son, Kook; Yang, Jiha; Kim, Sung Hak; Lee, Chang-Hun

2013-09-01

Fanconi anemia (FA) proteins are known to play roles in the cellular response to DNA interstrand cross-linking lesions; however, several reports have suggested that FA proteins play additional roles. To elucidate novel functions of FA proteins, we used yeast two-hybrid screening to identify binding partners of the Fanconi anemia complementation group A (FANCA) protein. The candidate proteins included never-in-mitosis-gene A (NIMA)-related kinase 2 (Nek2), which functions in the maintenance of centrosome integrity. The interaction of FANCA and Nek2 was confirmed in human embryonic kidney (HEK) 293T cells. Furthermore, FANCA interacted with γ-tubulin and localized to centrosomes, most notably during the mitotic phase, confirming that FANCA is a centrosomal protein. Knockdown of FANCA increased the frequency of centrosomal abnormalities and enhanced the sensitivity of U2OS osteosarcoma cells to nocodazole, a microtubule-interfering agent. In vitro kinase assays indicated that Nek2 can phosphorylate FANCA at threonine-351 (T351), and analysis with a phospho-specific antibody confirmed that this phosphorylation occurred in response to nocodazole treatment. Furthermore, U2OS cells overexpressing the phosphorylation-defective T351A FANCA mutant showed numerical centrosomal abnormalities, aberrant mitotic arrest, and enhanced nocodazole sensitivity, implying that the Nek2-mediated T351 phosphorylation of FANCA is important for the maintenance of centrosomal integrity. Taken together, this study revealed that FANCA localizes to centrosomes and is required for the maintenance of centrosome integrity, possibly through its phosphorylation at T351 by Nek2. Copyright © 2013 Elsevier Ltd. All rights reserved.

Interaction between ROCK II and nucleophosmin/B23 in the regulation of centrosome duplication.

PubMed

Ma, Zhiyong; Kanai, Masayuki; Kawamura, Kenji; Kaibuchi, Kozo; Ye, Keqiang; Fukasawa, Kenji

2006-12-01

Nucleophosmin (NPM)/B23 has been implicated in the regulation of centrosome duplication. NPM/B23 localizes between two centrioles in the unduplicated centrosome. Upon phosphorylation on Thr(199) by cyclin-dependent kinase 2 (CDK2)/cyclin E, the majority of centrosomal NPM/B23 dissociates from centrosomes, but some NPM/B23 phosphorylated on Thr(199) remains at centrosomes. It has been shown that Thr(199) phosphorylation of NPM/B23 is critical for the physical separation of the paired centrioles, an initial event of the centrosome duplication process. Here, we identified ROCK II kinase, an effector of Rho small GTPase, as a protein that localizes to centrosomes and physically interacts with NPM/B23. Expression of the constitutively active form of ROCK II promotes centrosome duplication, while down-regulation of ROCK II expression results in the suppression of centrosome duplication, especially delaying the initiation of centrosome duplication during the cell cycle. Moreover, ROCK II regulates centrosome duplication in its kinase and centrosome localization activity-dependent manner. We further found that ROCK II kinase activity is significantly enhanced by binding to NPM/B23 and that NPM/B23 acquires a higher binding affinity to ROCK II upon phosphorylation on Thr(199). Moreover, physical interaction between ROCK II and NPM/B23 in vivo occurs in association with CDK2/cyclin E activation and the emergence of Thr(199)-phosphorylated NPM/B23. All these findings point to ROCK II as the effector of the CDK2/cyclin E-NPM/B23 pathway in the regulation of centrosome duplication.

Oncogene-like induction of cellular invasion from centrosome amplification

PubMed Central

Godinho, Susana A.; Picone, Remigio; Burute, Mithila; Dagher, Regina; Su, Ying; Leung, Cheuk T.; Polyak, Kornelia; Brugge, Joan S.; Thery, Manuel; Pellman, David

2014-01-01

Centrosome amplification has long been recognized as a feature of human tumors, however its role in tumorigenesis remains unclear1. Centrosome amplification is poorly tolerated by non-transformed cells, and, in the absence of selection, extra centrosomes are spontaneously lost2. Thus, the high frequency of centrosome amplification, particularly in more aggressive tumors3, raises the possibility that extra centrosomes could, in some contexts, confer advantageous characteristics that promote tumor progression. Using a three-dimensional model system and other approaches to culture human mammary epithelial cells, we find that centrosome amplification triggers cell invasion. This invasive behavior is similar to that induced by overexpression of the breast cancer oncogene ErbB24 and indeed enhances invasiveness triggered by ErbB2. We show that, through increased centrosomal microtubule nucleation, centrosome amplification increases Rac1 activity, which disrupts normal cell-cell adhesion and promotes invasion. These findings demonstrate that centrosome amplification, a structural alteration of the cytoskeleton, can promote features of malignant transformation. PMID:24739973

Stable centrosomal roots disentangle to allow interphase centriole independence

PubMed Central

2018-01-01

The centrosome is a non–membrane-bound cellular compartment consisting of 2 centrioles surrounded by a protein coat termed the pericentriolar material (PCM). Centrioles generally remain physically associated together (a phenomenon called centrosome cohesion), yet how this occurs in the absence of a bounding lipid membrane is unclear. One model posits that pericentriolar fibres formed from rootletin protein directly link centrioles, yet little is known about the structure, biophysical properties, or assembly kinetics of such fibres. Here, I combine live-cell imaging of endogenously tagged rootletin with cell fusion and find previously unrecognised plasticity in centrosome cohesion. Rootletin forms large, diffusionally stable bifurcating fibres, which amass slowly on mature centrioles over many hours from anaphase. Nascent centrioles (procentrioles), in contrast, do not form roots and must be licensed to do so through polo-like kinase 1 (PLK1) activity. Transient separation of roots accompanies centriolar repositioning during the interphase, suggesting that centrioles organize as independent units, each containing discrete roots. Indeed, forced induction of duplicate centriole pairs allows independent reshuffling of individual centrioles between the pairs. Therefore collectively, these findings suggest that progressively nucleated polymers mediate the dynamic association of centrioles as either 1 or 2 interphase centrosomes, with implications for the understanding of how non–membrane-bound organelles self-organise. PMID:29649211

Polo-like kinase 1 regulates Nlp, a centrosome protein involved in microtubule nucleation.

PubMed

Casenghi, Martina; Meraldi, Patrick; Weinhart, Ulrike; Duncan, Peter I; Körner, Roman; Nigg, Erich A

2003-07-01

In animal cells, most microtubules are nucleated at centrosomes. At the onset of mitosis, centrosomes undergo a structural reorganization, termed maturation, which leads to increased microtubule nucleation activity. Centrosome maturation is regulated by several kinases, including Polo-like kinase 1 (Plk1). Here, we identify a centrosomal Plk1 substrate, termed Nlp (ninein-like protein), whose properties suggest an important role in microtubule organization. Nlp interacts with two components of the gamma-tubulin ring complex and stimulates microtubule nucleation. Plk1 phosphorylates Nlp and disrupts both its centrosome association and its gamma-tubulin interaction. Overexpression of an Nlp mutant lacking Plk1 phosphorylation sites severely disturbs mitotic spindle formation. We propose that Nlp plays an important role in microtubule organization during interphase, and that the activation of Plk1 at the onset of mitosis triggers the displacement of Nlp from the centrosome, allowing the establishment of a mitotic scaffold with enhanced microtubule nucleation activity.

Rho-kinase signaling controls nucleocytoplasmic shuttling of class IIa Histone Deacetylase (HDAC7) and transcriptional activation of orphan nuclear receptor NR4A1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Compagnucci, Claudia; Barresi, Sabina; Petrini, Stefania

2015-04-03

Rho-kinase (ROCK) has been well documented to play a key role in RhoA-induced actin remodeling. ROCK activation results in myosin light chain (MLC) phosphorylation either by direct action on MLC kinase (MLCK) or by inhibition of MLC phosphatase (MLCP), modulating actin–myosin contraction. We found that inhibition of the ROCK pathway in induced pluripotent stem cells, leads to nuclear export of HDAC7 and transcriptional activation of the orphan nuclear receptor NR4A1 while in cells with constitutive ROCK hyperactivity due to loss of function of the RhoGTPase activating protein Oligophrenin-1 (OPHN1), the orphan nuclear receptor NR4A1 is downregulated. Our study identify amore » new target of ROCK signaling via myosin phosphatase subunit (MYPT1) and Histone Deacetylase (HDAC7) at the nuclear level and provide new insights in the cellular functions of ROCK. - Highlights: • ROCK regulates nucleocytoplasmic shuttling of HDAC7 via phosphorylation of MYPT1. • Nuclear export of HDAC7 and upregulation of NR4A1 occurs with low ROCK activity. • High levels of ROCK activity due to OPHN1 loss of function downregulate NR4A1.« less

Is cohesin required for spindle-pole-body/centrosome cohesion?

PubMed Central

Jin, Hui; Avey, Martin

2012-01-01

Centrosomes are microtubule-organizing centers that nucleate spindle microtubules during cell division. In budding yeast, the centrosome, often referred to as the spindle pole body, shares structural components with the centriole, the central core of the animal centrosome. The parental centrosome is duplicated when DNA replication takes place. Like sister chromatids tethered together by cohesin, duplicated centrosomes are linked and then separate to form the bipolar spindle necessary for chromosome segregation. Recent studies have shown that cohesin is also localized to the animal centrosome and is perhaps directly involved in engaging paired centrioles. Here we discuss the potential role of cohesin in mediating spindle-pole-body cohesion in the context of yeast meiosis. We propose that the coordination of chromosome segregation with centrosome cohesion and duplication is mediated by the antagonistic interaction between the Aurora kinase and the Polo kinase and that the role of cohesin in centrosome regulation appears to be indirect in budding yeast. PMID:22482005

Loss and Rebirth of the Animal Microtubule Organizing Center: How Maternal Expression of Centrosomal Proteins Cooperates with the Sperm Centriole in Zygotic Centrosome Reformation.

PubMed

Inoue, Daigo; Wittbrodt, Joachim; Gruss, Oliver J

2018-04-01

Centrosomes are the main microtubule organizing centers in animal cells. In particular during embryogenesis, they ensure faithful spindle formation and proper cell divisions. As metazoan centrosomes are eliminated during oogenesis, they have to be reassembled upon fertilization. Most metazoans use the sperm centrioles as templates for new centrosome biogenesis while the egg's cytoplasm re-prepares all components for on-going centrosome duplication in rapidly dividing embryonic cells. We discuss our knowledge and the experimental challenges to analyze zygotic centrosome reformation, which requires genetic experiments to enable scrutinizing respective male and female contributions. Male and female knockout animals and mRNA injection to mimic maternal expression of centrosomal proteins could point a way to the systematic molecular dissection of the process. The most recent data suggest that timely expression of centrosome components in oocytes is the key to zygotic centrosome reformation that uses male sperm as coordinators for de novo centrosome production. © 2018 WILEY Periodicals, Inc.

Ndj1, a telomere-associated protein, regulates centrosome separation in budding yeast meiosis

PubMed Central

Li, Ping; Shao, Yize; Jin, Hui

2015-01-01

Yeast centrosomes (called spindle pole bodies [SPBs]) remain cohesive for hours during meiotic G2 when recombination takes place. In contrast, SPBs separate within minutes after duplication in vegetative cells. We report here that Ndj1, a previously known meiosis-specific telomere-associated protein, is required for protecting SPB cohesion. Ndj1 localizes to the SPB but dissociates from it ∼16 min before SPB separation. Without Ndj1, meiotic SPBs lost cohesion prematurely, whereas overproduction of Ndj1 delayed SPB separation. When produced ectopically in vegetative cells, Ndj1 caused SPB separation defects and cell lethality. Localization of Ndj1 to the SPB depended on the SUN domain protein Mps3, and removal of the N terminus of Mps3 allowed SPB separation and suppressed the lethality of NDJ1-expressing vegetative cells. Finally, we show that Ndj1 forms oligomeric complexes with Mps3, and that the Polo-like kinase Cdc5 regulates Ndj1 protein stability and SPB separation. These findings reveal the underlying mechanism that coordinates yeast centrosome dynamics with meiotic telomere movement and cell cycle progression. PMID:25897084

Small organelle, big responsibility: the role of centrosomes in development and disease

PubMed Central

Chavali, Pavithra L.; Pütz, Monika; Gergely, Fanni

2014-01-01

The centrosome, a key microtubule organizing centre, is composed of centrioles, embedded in a protein-rich matrix. Centrosomes control the internal spatial organization of somatic cells, and as such contribute to cell division, cell polarity and migration. Upon exiting the cell cycle, most cell types in the human body convert their centrioles into basal bodies, which drive the assembly of primary cilia, involved in sensing and signal transduction at the cell surface. Centrosomal genes are targeted by mutations in numerous human developmental disorders, ranging from diseases exclusively affecting brain development, through global growth failure syndromes to diverse pathologies associated with ciliary malfunction. Despite our much-improved understanding of centrosome function in cellular processes, we know remarkably little of its role in the organismal context, especially in mammals. In this review, we examine how centrosome dysfunction impacts on complex physiological processes and speculate on the challenges we face when applying knowledge generated from in vitro and in vivo model systems to human development. PMID:25047622

[Hyperoxia induces reactive oxygen species production and promotes SIRT1 nucleocytoplasmic shuttling of peripheral blood mononuclear cells in premature infants in vitro].

PubMed

Yang, Xi; Dong, Wenbin; Li, Qingping; Kang, Lan; Lei, Xiaoping; Zhang, Lianyu; Lu, Youying; Zhai, Xuesong

2015-12-01

To explore the relationship between deacetylase sirtuin 1 (SIRT1) and reactive oxygen species (ROS) after oxygen therapy in the peripheral blood mononuclear cells (PBMCs) of the premature infants. According to the fraction of inspired O2 (FiO2), premature infants diagnosed with respiratory distress syndrome (RDS) (gestational age <32 weeks), were divided into three groups: low dosage oxygen group (FiO2 <300 mL/L), moderate dosage oxygen group (FiO2; 300 mL/L-400 mL/L), high dosage oxygen group (FiO2 >400 mL/L). After 48 hours of oxygen treatment, PBMCs and serum were collected from the peripheral blood. Then the intracellular ROS level was detected by MitoSOX(TM) Red labeling combined with confocal laser scanning microscopy; the malondialdehyde (MDA) content in the serum was determined by the whole spectrum spectrophotometer; the SIRT1 localization was observed by immunofluorescence staining; and the SIRT1 levels in PBMCs were examined by Western blotting. With the increase of FiO2, the ROS, MDA content and the rate of SIRT1 nucleocytoplasmic shuttling of PBMCs gradually increased and SIRT1 protein expression was significantly lowered. Hyperoxia induces ROS production in premature infants, promotes SIRT1 to cross from nucleus to cytoplasm, inhibits the resistant ability of SIRT1 to oxidative stress.

Mps1 as a link between centrosomes and genomic instability.

PubMed

Kasbek, Christopher; Yang, Ching-Hui; Fisk, Harold A

2009-10-01

Centrosomes are microtubule-organizing centers that must be precisely duplicated before mitosis. Centrosomes regulate mitotic spindle assembly, and the presence of excess centrosomes leads to the production of aberrant mitotic spindles which generate chromosome segregation errors. Many human tumors possess excess centrosomes that lead to the production of abnormal spindles in situ. In some tumors, these extra centrosomes appear before aneuploidy, suggesting that defects in centrosome duplication might promote genomic instability and tumorigenesis. The Mps1 protein kinase is required for centrosome duplication, and preventing the proteasome-dependent degradation of Mps1 at centrosomes increases its local concentration and causes the production of excess centrosomes during a prolonged S-phase. Here, we show that Mps1 degradation is misregulated in two tumor-derived cell lines, and that the failure to appropriately degrade Mps1 correlates with the ability of these cells to produce extra centrosomes during a prolonged S-phase. In the 21NT breast-tumor derived cell line, a mutant Mps1 protein that is normally constitutively degraded can accumulate at centrosomes and perturb centrosome duplication, suggesting that these cells have a defect in the mechanisms that target Mps1 to the proteasome. In contrast, the U2OS osteosarcoma cell line expresses a nondegradable form of Mps1, which we show causes the dose-dependent over duplication of centrioles even at very low levels of expression. Our data demonstrate that defects in Mps1 degradation can occur through multiple mechanisms, and suggest that Mps1 may provide a link between the control of centrosome duplication and genomic instability. (c) 2009 Wiley-Liss, Inc.

Structure and non-structure of centrosomal proteins.

PubMed

Dos Santos, Helena G; Abia, David; Janowski, Robert; Mortuza, Gulnahar; Bertero, Michela G; Boutin, Maïlys; Guarín, Nayibe; Méndez-Giraldez, Raúl; Nuñez, Alfonso; Pedrero, Juan G; Redondo, Pilar; Sanz, María; Speroni, Silvia; Teichert, Florian; Bruix, Marta; Carazo, José M; Gonzalez, Cayetano; Reina, José; Valpuesta, José M; Vernos, Isabelle; Zabala, Juan C; Montoya, Guillermo; Coll, Miquel; Bastolla, Ugo; Serrano, Luis

2013-01-01

Here we perform a large-scale study of the structural properties and the expression of proteins that constitute the human Centrosome. Centrosomal proteins tend to be larger than generic human proteins (control set), since their genes contain in average more exons (20.3 versus 14.6). They are rich in predicted disordered regions, which cover 57% of their length, compared to 39% in the general human proteome. They also contain several regions that are dually predicted to be disordered and coiled-coil at the same time: 55 proteins (15%) contain disordered and coiled-coil fragments that cover more than 20% of their length. Helices prevail over strands in regions homologous to known structures (47% predicted helical residues against 17% predicted as strands), and even more in the whole centrosomal proteome (52% against 7%), while for control human proteins 34.5% of the residues are predicted as helical and 12.8% are predicted as strands. This difference is mainly due to residues predicted as disordered and helical (30% in centrosomal and 9.4% in control proteins), which may correspond to alpha-helix forming molecular recognition features (α-MoRFs). We performed expression assays for 120 full-length centrosomal proteins and 72 domain constructs that we have predicted to be globular. These full-length proteins are often insoluble: Only 39 out of 120 expressed proteins (32%) and 19 out of 72 domains (26%) were soluble. We built or retrieved structural models for 277 out of 361 human proteins whose centrosomal localization has been experimentally verified. We could not find any suitable structural template with more than 20% sequence identity for 84 centrosomal proteins (23%), for which around 74% of the residues are predicted to be disordered or coiled-coils. The three-dimensional models that we built are available at http://ub.cbm.uam.es/centrosome/models/index.php.

Breaking the ties that bind: new advances in centrosome biology.

PubMed

Mardin, Balca R; Schiebel, Elmar

2012-04-02

The centrosome, which consists of two centrioles and the surrounding pericentriolar material, is the primary microtubule-organizing center (MTOC) in animal cells. Like chromosomes, centrosomes duplicate once per cell cycle and defects that lead to abnormalities in the number of centrosomes result in genomic instability, a hallmark of most cancer cells. Increasing evidence suggests that the separation of the two centrioles (disengagement) is required for centrosome duplication. After centriole disengagement, a proteinaceous linker is established that still connects the two centrioles. In G2, this linker is resolved (centrosome separation), thereby allowing the centrosomes to separate and form the poles of the bipolar spindle. Recent work has identified new players that regulate these two processes and revealed unexpected mechanisms controlling the centrosome cycle.

Small organelle, big responsibility: the role of centrosomes in development and disease.

PubMed

Chavali, Pavithra L; Pütz, Monika; Gergely, Fanni

2014-09-05

The centrosome, a key microtubule organizing centre, is composed of centrioles, embedded in a protein-rich matrix. Centrosomes control the internal spatial organization of somatic cells, and as such contribute to cell division, cell polarity and migration. Upon exiting the cell cycle, most cell types in the human body convert their centrioles into basal bodies, which drive the assembly of primary cilia, involved in sensing and signal transduction at the cell surface. Centrosomal genes are targeted by mutations in numerous human developmental disorders, ranging from diseases exclusively affecting brain development, through global growth failure syndromes to diverse pathologies associated with ciliary malfunction. Despite our much-improved understanding of centrosome function in cellular processes, we know remarkably little of its role in the organismal context, especially in mammals. In this review, we examine how centrosome dysfunction impacts on complex physiological processes and speculate on the challenges we face when applying knowledge generated from in vitro and in vivo model systems to human development. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Ndj1, a telomere-associated protein, regulates centrosome separation in budding yeast meiosis.

PubMed

Li, Ping; Shao, Yize; Jin, Hui; Yu, Hong-Guo

2015-04-27

Yeast centrosomes (called spindle pole bodies [SPBs]) remain cohesive for hours during meiotic G2 when recombination takes place. In contrast, SPBs separate within minutes after duplication in vegetative cells. We report here that Ndj1, a previously known meiosis-specific telomere-associated protein, is required for protecting SPB cohesion. Ndj1 localizes to the SPB but dissociates from it ∼16 min before SPB separation. Without Ndj1, meiotic SPBs lost cohesion prematurely, whereas overproduction of Ndj1 delayed SPB separation. When produced ectopically in vegetative cells, Ndj1 caused SPB separation defects and cell lethality. Localization of Ndj1 to the SPB depended on the SUN domain protein Mps3, and removal of the N terminus of Mps3 allowed SPB separation and suppressed the lethality of NDJ1-expressing vegetative cells. Finally, we show that Ndj1 forms oligomeric complexes with Mps3, and that the Polo-like kinase Cdc5 regulates Ndj1 protein stability and SPB separation. These findings reveal the underlying mechanism that coordinates yeast centrosome dynamics with meiotic telomere movement and cell cycle progression. © 2015 Li et al.

The Centrosome and Its Duplication Cycle

PubMed Central

Fu, Jingyan; Hagan, Iain M.; Glover, David M.

2015-01-01

The centrosome was discovered in the late 19th century when mitosis was first described. Long recognized as a key organelle of the spindle pole, its core component, the centriole, was realized more than 50 or so years later also to comprise the basal body of the cilium. Here, we chart the more recent acquisition of a molecular understanding of centrosome structure and function. The strategies for gaining such knowledge were quickly developed in the yeasts to decipher the structure and function of their distinctive spindle pole bodies. Only within the past decade have studies with model eukaryotes and cultured cells brought a similar degree of sophistication to our understanding of the centrosome duplication cycle and the multiple roles of this organelle and its component parts in cell division and signaling. Now as we begin to understand these functions in the context of development, the way is being opened up for studies of the roles of centrosomes in human disease. PMID:25646378

Nucleocytoplasmic transport in cells with progerin-induced defective nuclear lamina.

PubMed

Ferri, Gianmarco; Storti, Barbara; Bizzarri, Ranieri

2017-10-01

Recent data indicate that nuclear lamina (NL) plays a relevant role in many fundamental cellular functions. The peculiar role of NL in cells is dramatically demonstrated by the Hutchinson-Gilford progeria syndrome (HGPS), an inherited laminopathy that causes premature, rapid aging shortly after birth. In HGPS, a mutant form of Lamin A (progeria) leads to a dysmorphic NL structure, but how this perturbation is transduced into cellular changes is still largely unknown. Owing to the close structural relationship between NL and the Nuclear Pore Complex (NPC), in this work we test whether HGPS affects passive and active nucleo-cytoplasmic shuttling of cargoes by means of an established model based of fluorescence recovery after photobleaching. Our findings clearly demonstrate that dysmorphic NL is decoupled from the dynamic characteristics of passive and active transport towards and from the nucleus, as well as from the binding affinity of transport protein mediators. Copyright © 2017 Elsevier B.V. All rights reserved.

Increased centrosome amplification in aged stem cells of the Drosophila midgut

DOE Office of Scientific and Technical Information (OSTI.GOV)

Park, Joung-Sun; Pyo, Jung-Hoon; Na, Hyun-Jin

Highlights: • Increased centrosome amplification in ISCs of aged Drosophila midguts. • Increased centrosome amplification in ISCs of oxidative stressed Drosophila midguts. • Increased centrosome amplification in ISCs by overexpression of PVR, EGFR, and AKT. • Supernumerary centrosomes can be responsible for abnormal ISC polyploid cells. • Supernumerary centrosomes can be a useful marker for aging stem cells. - Abstract: Age-related changes in long-lived tissue-resident stem cells may be tightly linked to aging and age-related diseases such as cancer. Centrosomes play key roles in cell proliferation, differentiation and migration. Supernumerary centrosomes are known to be an early event in tumorigenesismore » and senescence. However, the age-related changes of centrosome duplication in tissue-resident stem cells in vivo remain unknown. Here, using anti-γ-tubulin and anti-PH3, we analyzed mitotic intestinal stem cells with supernumerary centrosomes in the adult Drosophila midgut, which may be a versatile model system for stem cell biology. The results showed increased centrosome amplification in intestinal stem cells of aged and oxidatively stressed Drosophila midguts. Increased centrosome amplification was detected by overexpression of PVR, EGFR, and AKT in intestinal stem cells/enteroblasts, known to mimic age-related changes including hyperproliferation of intestinal stem cells and hyperplasia in the midgut. Our data show the first direct evidence for the age-related increase of centrosome amplification in intestinal stem cells and suggest that the Drosophila midgut is an excellent model for studying molecular mechanisms underlying centrosome amplification in aging adult stem cells in vivo.« less

Centrosomes are autocatalytic droplets of pericentriolar material organized by centrioles

NASA Astrophysics Data System (ADS)

Zwicker, David; Decker, Markus; Jaensch, Steffen; Hyman, Anthony A.; Jülicher, Frank

2014-03-01

We propose a physical description of the centrosome, a membrane-less organelle involved in cell division. In our model, centrosome material occurs in a soluble form in the cytosol and a form that tends to undergo phase separation from the cytosol. We find that an autocatalytic chemical transition between these forms accounts for the temporal evolution observed in experiments. Interestingly, the nucleation of centrosomes can be controlled by an enzymatic activity of the centrioles, which are present at the core of all centrosomes. This non-equilibrium feature also allows for multiple stable centrosomes, a situation which is unstable in equilibrium phase separation. Our theory explains the growth dynamics of centrosomes for all cell sizes down to the eight-cell stage of the C. elegans embryo. It also accounts for data acquired in experiments with aberrant numbers of centrosomes and altered cell volumes. Furthermore, our model can describe unequal centrosome sizes observed in cells with disturbed centrioles. Our example suggests a general picture of the organization of membrane-less organelles.

Force-balance model of suppression of multipolar division in cancer cells with extra centrosomes

NASA Astrophysics Data System (ADS)

Zhu, Jie

2013-03-01

Cancer cells often possess extra centrosomes which have the potential to cause cell death due to catastrophic multipolar division. Many cancer cells, however, are able to escape multipolar mitosis by clustering the extra centrosomes to form bipolar spindles. The mechanism of centrosome clustering is therefore of great interest to the development of anti-cancer drugs because the de-clustering of extra centrosomes provides an appealing way to eliminate cancer cells while keeping healthy cells intact. We present a physical model assuming 1) dynamic centrosomal microtubules interact with chromosomes by both pushing on chromosome arms and pulling along kinetochores; 2) these microtubules interact with force generators associated with actin/adhesion structures at the cell boundary; and 3) motors act on anti-parallel microtubules from different centrosomes. We find via computer simulations that chromosomes tend to aggregate near the cell center while centrosomes can be either clustered to form bipolar spindles or scattered to form multipolar spindles, depending on the strengths of relative forces, cell shape and adhesion geometry. The model predictions agree with data from cells plated on adhesive micropatterns and from biochemically or genetically perturbed cells. Furthermore, our model is able to explain various microtubule distributions in interphase cells on patterned substrates. This work was supported by NSF

Centrosomes are autocatalytic droplets of pericentriolar material organized by centrioles

PubMed Central

Zwicker, David; Decker, Markus; Jaensch, Steffen; Hyman, Anthony A.; Jülicher, Frank

2014-01-01

Centrosomes are highly dynamic, spherical organelles without a membrane. Their physical nature and their assembly are not understood. Using the concept of phase separation, we propose a theoretical description of centrosomes as liquid droplets. In our model, centrosome material occurs in a form soluble in the cytosol and a form that tends to undergo phase separation from the cytosol. We show that an autocatalytic chemical transition between these forms accounts for the temporal evolution observed in experiments. Interestingly, the nucleation of centrosomes can be controlled by an enzymatic activity of the centrioles, which are present at the core of all centrosomes. This nonequilibrium feature also allows for multiple stable centrosomes, a situation that is unstable in equilibrium phase separation. Our theory explains the growth dynamics of centrosomes for all cell sizes down to the eight-cell stage of the Caenorhabditis elegans embryo, and it also accounts for data acquired in experiments with aberrant numbers of centrosomes and altered cell volumes. Furthermore, the model can describe unequal centrosome sizes observed in cells with perturbed centrioles. We also propose an interpretation of the molecular details of the involved proteins in the case of C. elegans. Our example suggests a general picture of the organization of membraneless organelles. PMID:24979791

Centrosomes are autocatalytic droplets of pericentriolar material organized by centrioles.

PubMed

Zwicker, David; Decker, Markus; Jaensch, Steffen; Hyman, Anthony A; Jülicher, Frank

2014-07-01

Centrosomes are highly dynamic, spherical organelles without a membrane. Their physical nature and their assembly are not understood. Using the concept of phase separation, we propose a theoretical description of centrosomes as liquid droplets. In our model, centrosome material occurs in a form soluble in the cytosol and a form that tends to undergo phase separation from the cytosol. We show that an autocatalytic chemical transition between these forms accounts for the temporal evolution observed in experiments. Interestingly, the nucleation of centrosomes can be controlled by an enzymatic activity of the centrioles, which are present at the core of all centrosomes. This nonequilibrium feature also allows for multiple stable centrosomes, a situation that is unstable in equilibrium phase separation. Our theory explains the growth dynamics of centrosomes for all cell sizes down to the eight-cell stage of the Caenorhabditis elegans embryo, and it also accounts for data acquired in experiments with aberrant numbers of centrosomes and altered cell volumes. Furthermore, the model can describe unequal centrosome sizes observed in cells with perturbed centrioles. We also propose an interpretation of the molecular details of the involved proteins in the case of C. elegans. Our example suggests a general picture of the organization of membraneless organelles.

Heterodimerization with Jun family members regulates c-Fos nucleocytoplasmic traffic.

PubMed

Malnou, Cécile E; Salem, Tamara; Brockly, Frédérique; Wodrich, Harald; Piechaczyk, Marc; Jariel-Encontre, Isabelle

2007-10-19

c-Fos proto-oncoprotein forms AP-1 transcription complexes with heterodimerization partners such as c-Jun, JunB, and JunD. Thereby, it controls essential cell functions and exerts tumorigenic actions. The dynamics of c-Fos intracellular distribution is poorly understood. Hence, we have combined genetic, cell biology, and microscopic approaches to investigate this issue. In addition to a previously characterized basic nuclear localization signal (NLS) located within the central DNA-binding domain, we identified a second NLS within the c-Fos N-terminal region. This NLS is non-classic and its activity depends on transportin 1 in vivo. Under conditions of prominent nuclear localization, c-Fos can undergo nucleocytoplasmic shuttling through an active Crm-1 exportin-independent mechanism. Dimerization with the Jun proteins inhibits c-Fos nuclear exit. The strongest effect is observed with c-Jun probably in accordance with the relative stabilities of the different c-Fos:Jun dimers. Retrotransport inhibition is not caused by binding of dimers to DNA and, therefore, is not induced by indirect effects linked to activation of c-Fos target genes. Monomeric, but not dimeric, Jun proteins also shuttle actively. Thus, our work unveils a novel regulation operating on AP-1 by demonstrating that dimerization is crucial, not only for active transcription complex formation, but also for keeping them in the compartment where they exert their transcriptional function.

The centrosomal component CEP161 of Dictyostelium discoideum interacts with the Hippo signaling pathway

PubMed Central

Sukumaran, Salil K.; Blau-Wasser, Rosemarie; Rohlfs, Meino; Gallinger, Christoph; Schleicher, Michael; Noegel, Angelika A

2015-01-01

CEP161 is a novel component of the Dictyostelium discoideum centrosome which was identified as binding partner of the pericentriolar component CP250. Here we show that the amino acids 1-763 of the 1381 amino acids CEP161 are sufficient for CP250 binding, centrosomal targeting and centrosome association. Analysis of AX2 cells over-expressing truncated and full length CEP161 proteins revealed defects in growth and development. By immunoprecipitation experiments we identified the Hippo related kinase SvkA (Hrk-svk) as binding partner for CEP161. Both proteins colocalize at the centrosome. In in vitro kinase assays the N-terminal domain of CEP161 (residues 1-763) inhibited the kinase activity of Hrk-svk. A comparison of D. discoideum Hippo kinase mutants with mutants overexpressing CEP161 polypeptides revealed similar defects. We propose that the centrosomal component CEP161 is a novel player in the Hippo signaling pathway and affects various cellular properties through this interaction. PMID:25607232

Centrosome centering and decentering by microtubule network rearrangement

PubMed Central

Letort, Gaëlle; Nedelec, Francois; Blanchoin, Laurent; Théry, Manuel

2016-01-01

The centrosome is positioned at the cell center by pushing and pulling forces transmitted by microtubules (MTs). Centrosome decentering is often considered to result from asymmetric, cortical pulling forces exerted in particular by molecular motors on MTs and controlled by external cues affecting the cell cortex locally. Here we used numerical simulations to investigate the possibility that it could equally result from the redistribution of pushing forces due to a reorientation of MTs. We first showed that MT gliding along cell edges and pivoting around the centrosome regulate MT rearrangement and thereby direct the spatial distribution of pushing forces, whereas the number, dynamics, and stiffness of MTs determine the magnitude of these forces. By modulating these parameters, we identified different regimes, involving both pushing and pulling forces, characterized by robust centrosome centering, robust off-centering, or “reactive” positioning. In the last-named conditions, weak asymmetric cues can induce a misbalance of pushing and pulling forces, resulting in an abrupt transition from a centered to an off-centered position. Taken together, these results point to the central role played by the configuration of the MTs on the distribution of pushing forces that position the centrosome. We suggest that asymmetric external cues should not be seen as direct driver of centrosome decentering and cell polarization but instead as inducers of an effective reorganization of the MT network, fostering centrosome motion to the cell periphery. PMID:27440925

Centrosome Clustering in the Development of Bovine Binucleate Trophoblast Giant Cells.

PubMed

Klisch, Karl; Schraner, Elisabeth M; Boos, Alois

2017-01-01

Binucleate trophoblast giant cells (BNC) are the characteristic feature of the ruminant placenta. During their development, BNC pass through 2 acytokinetic mitoses and become binucleate with 2 tetraploid nuclei. In this study, we investigate the number and location of centrosomes in bovine BNC. Centrosomes typically consist of 2 centrioles surrounded by electron-dense pericentriolar material. Duplication of centrosomes is tightly linked to the cell cycle, which ensures that the number of centrosomes remains constant in proliferating diploid cells. Alterations of the cell cycle, which affect the number of chromosome sets, also affect the number of centrosomes. In this study, we use placentomal tissue from pregnant cows (gestational days 80-230) for immunohistochemical staining of γ-tubulin (n = 3) and transmission electron microscopy (n = 3). We show that mature BNC have 4 centrosomes with 8 centrioles, clustered in the angle between the 2 cell nuclei. During the second acytokinetic mitosis, the centrosomes must be clustered to form the poles of a bipolar spindle. In rare cases, centrosome clustering fails and tripolar mitosis leads to the formation of trinucleate "BNC". Generally, centrosome clustering occurs in polyploid tumor cells, which have an increased number of centrioles, but it is absent in proliferating diploid cells. Thus, inhibition of centrosome clustering in tumor cells is a novel promising strategy for cancer treatment. BNC are a cell population in which centrosome clustering occurs as part of the normal life history. Thus, they might be a good model for the study of the molecular mechanisms of centrosome clustering. © 2016 S. Karger AG, Basel.

Cell lines derived from feline fibrosarcoma display unstable chromosomal aneuploidy and additionally centrosome number aberrations.

PubMed

von Erichsen, J; Hecht, W; Löhberg-Gruene, C; Reinacher, M

2012-07-01

The purpose of the study was to evaluate clonality and presence of numerical chromosomal and centrosomal aberrations in 5 established feline fibrosarcoma cell lines and in a fetal dermal fibroblast cell line as a control. The clonality of all cell lines was examined using limited-dilution cloning. The number of chromosomes was counted in metaphase spreads. The immunocytochemical analysis of centrosome numbers was performed by indirect immunofluorescence using a monoclonal antibody that targets γ-tubulin, a well-characterized component of centrosomes. Monoclonal cell populations could be established from all cell lines. In all feline fibrosarcoma cell lines, the number of chromosomes deviated abnormally from the normal feline chromosome number of 2n = 38, ranging from 19 to 155 chromosomes per cell. Centrosome hyperamplification was observed in all 5 feline fibrosarcoma cell lines with a proportion of cells (5.7 to 15.2%) having more than 2 centrosomes. In the control cell line, only 0.6% of the cells had more than 2 centrosomes. In conclusion, the examinations revealed that centrosome hyperamplification occurs in feline fibrosarcoma cell lines. The feline fibrosarcoma cell lines possessed 10 to 25 times as many cells with centrosome hyperamplification as the control cell line. These observations suggest an association of numerical centrosome aberrations with karyotype instability by increasing the frequency of chromosome missegregation. The results of this study may be helpful for further characterization of feline fibrosarcomas and may contribute to the knowledge of cytogenetic factors that may be important for the pathogenesis of feline fibrosarcomas.

Stabilization of cartwheel-less centrioles for duplication requires CEP295-mediated centriole to centrosome conversion

PubMed Central

Izquierdo, Denisse; Wang, Won-Jing; Uryu, Kunihiro; Tsou, Meng-Fu Bryan

2014-01-01

SUMMARY Vertebrate centrioles lose their geometric scaffold, the cartwheel, during mitosis, concurrently with gaining the ability to recruit the pericentriolar material (PCM) and thereby function as the centrosome. Cartwheel removal has recently been implicated in centriole duplication, but whether “cartwheel-less” centrioles are intrinsically stable, or must be maintained through other modifications remains unclear. Here, we identify a newborn centriole-enriched protein, KIAA1731/CEP295, specifically mediating centriole-to-centrosome conversion but dispensable for cartwheel removal. In the absence of CEP295, centrioles form in S/G2 phase, and lose their associated cartwheel in mitosis, but cannot be converted to centrosomes, uncoupling the two events. Strikingly, centrioles devoid of both the PCM and cartwheel progressively lose centriolar components, whereas centrioles associating with either the cartwheel or PCM alone can exist stably. Thus, cartwheel removal can have grave repercussions to centriole stability, and centriole-to-centrosome conversion mediated by CEP295 must occur in parallel to maintain cartwheel-less centrioles for duplication. PMID:25131205

Centrosome Linker-induced Tetraploid Segregation Errors Link Rhabdoid Phenotypes and Lethal Colorectal Cancers.

PubMed

Remo, Andrea; Manfrin, Erminia; Parcesepe, Pietro; Ferrarini, Alberto; Han, Hye Seung; Ugnius, Mickys; Laudanna, Carmelo; Simbolo, Michele; Malanga, Donatella; Mendes Oliveira, Duarte; Baritono, Elisabetta; Colangelo, Tommaso; Sabatino, Lina; Giuliani, Jacopo; Molinari, Enrico; Garonzi, Marianna; Xumerle, Luciano; Delledonne, Massimo; Giordano, Guido; Ghimenton, Claudio; Lonardo, Fortunato; D'angelo, Fulvio; Grillo, Federica; Mastracci, Luca; Viglietto, Giuseppe; Ceccarelli, Michele; Colantuoni, Vittorio; Scarpa, Aldo; Pancione, Massimo

2018-05-21

Centrosome anomalies contribute to tumorigenesis but it remains unclear how they are generated in lethal cancer phenotypes. Here, it is demonstrated that human microsatellite instable (MSI) and BRAF(V600E) mutant colorectal cancers with a lethal rhabdoid phenotype are characterized by inactivation of centrosomal functions. A splice site mutation that causes an unbalanced dosage of rootletin (CROCC), a centrosomal-linker component required for centrosome cohesion and separation at the chromosome 1p36.13 locus, resulted in abnormally shaped centrosomes in rhabdoid cells from human colon tissues. Notably, deleterious deletions at 1p36.13 were recurrent in a subgroup of BRAF(V600E) mutant and microsatellite stable (MSS) rhabdoid colorectal cancers but not in classical colorectal cancer or pediatric rhabdoid tumors. Interfering with CROCC expression in near-diploid BRAF(V600E) mutant/MSI colon cancer cells disrupts bipolar mitotic spindle architecture, promotes tetraploid segregation errors resulting in a highly aggressive rhabdoid-like phenotype in vitro. Restoring near-wild-type levels of CROCC in a metastatic model harboring 1p36.13 deletion results in correction of centrosome segregation errors and cell death, revealing a mechanism of tolerance to mitotic errors and tetraploidization promoted by deleterious 1p36.13 loss. Accordingly, cancer cells lacking 1p36.13 display far greater sensitivity to centrosome spindle pole stabilizing agents in vitro. These data shed light on a previously unknown link between centrosome cohesion defects and lethal cancer phenotypes providing new insight into pathways underlying genome instability. Mis-segregation of chromosomes is a prominent feature of chromosome instability and intra-tumoral heterogeneity recurrent in metastatic tumors for which the molecular basis is unknown. The present study provides insight into the mechanism by which defects in rootletin, a centrosome linker component causes tetraploid segregation errors and

Ligand-dependent nucleo-cytoplasmic shuttling of peroxisome proliferator-activated receptors, PPARα and PPARγ.

PubMed

Umemoto, Tomoe; Fujiki, Yukio

2012-07-01

Peroxisome proliferator-activated receptors (PPARs) play important roles in diverse biological processes including metabolisms of sugars and lipids and differentiation of cells such as adipocytes. PPARs are transcription factors belonging to the ligand-dependent hormone receptor group. To function as transcription factors, PPARs translocate into nucleus where they associate with transcription apparatus. However, mechanisms underlying nuclear transport of PPARs remain enigmatic. We show here that PPARα and PPARγ dynamically shuttle between nucleus and cytoplasm, although they constitutively and predominantly appear in nucleus. With a series of truncation mutants, we identify that PPAR nuclear transport is mediated by at least two nuclear localization signals (NLSs) in DNA-binding domain (DBD)-hinge and activation function 1 (AF1) regions and their respective receptors including importinα/β, importin 7, and an unidentified receptor. PPARs also harbor two nuclear export signals in DBD and ligand-binding domain regions that are recognized by distinct export receptors, calreticulin and CRM1. Moreover, we show that nuclear-cytoplasmic shuttling of PPARs is regulated by respective PPAR ligands and Ca2+ concentration. Taken together, we suggest that the multiple pathways for the nuclear-cytoplasmic transport of PPARs regulate the biological functions of PPARs in response to external signals. © 2012 The Authors Journal compilation © 2012 by the Molecular Biology Society of Japan/Blackwell Publishing Ltd.

Tank binding kinase 1 is a centrosome-associated kinase necessary for microtubule dynamics and mitosis

PubMed Central

Pillai, Smitha; Nguyen, Jonathan; Johnson, Joseph; Haura, Eric; Coppola, Domenico; Chellappan, Srikumar

2015-01-01

TANK Binding Kinase 1 (TBK1) is a non-canonical IκB kinase that contributes to KRAS-driven lung cancer. Here we report that TBK1 plays essential roles in mammalian cell division. Specifically, levels of active phospho-TBK1 increase during mitosis and localize to centrosomes, mitotic spindles and midbody, and selective inhibition or silencing of TBK1 triggers defects in spindle assembly and prevents mitotic progression. TBK1 binds to the centrosomal protein CEP170 and to the mitotic apparatus protein NuMA, and both CEP170 and NuMA are TBK1 substrates. Further, TBK1 is necessary for CEP170 centrosomal localization and binding to the microtubule depolymerase Kif2b, and for NuMA binding to dynein. Finally, selective disruption of the TBK1–CEP170 complex augments microtubule stability and triggers defects in mitosis, suggesting that TBK1 functions as a mitotic kinase necessary for microtubule dynamics and mitosis. PMID:26656453

Excess centrosomes perturb dynamic endothelial cell repolarization during blood vessel formation

PubMed Central

Kushner, Erich J.; Ferro, Luke S.; Yu, Zhixian; Bautch, Victoria L.

2016-01-01

Blood vessel formation requires dynamic movements of endothelial cells (ECs) within sprouts. The cytoskeleton regulates migratory polarity, and centrosomes organize the microtubule cytoskeleton. However, it is not well understood how excess centrosomes, commonly found in tumor stromal cells, affect microtubule dynamics and interphase cell polarity. Here we find that ECs dynamically repolarize during sprouting angiogenesis, and excess centrosomes block repolarization and reduce migration and sprouting. ECs with excess centrosomes initially had more centrosome-derived microtubules but, paradoxically, fewer steady-state microtubules. ECs with excess centrosomes had elevated Rac1 activity, and repolarization was rescued by blockade of Rac1 or actomyosin blockers, consistent with Rac1 activity promoting cortical retrograde actin flow and actomyosin contractility, which precludes cortical microtubule engagement necessary for dynamic repolarization. Thus normal centrosome numbers are required for dynamic repolarization and migration of sprouting ECs that contribute to blood vessel formation. PMID:27099371

Centrosome docking at the immunological synapse is controlled by Lck signaling

PubMed Central

Tsun, Andy; Qureshi, Ihjaaz; Stinchcombe, Jane C.; Jenkins, Misty R.; de la Roche, Maike; Kleczkowska, Joanna; Zamoyska, Rose

2011-01-01

Docking of the centrosome at the plasma membrane directs lytic granules to the immunological synapse. To identify signals controlling centrosome docking at the synapse, we have studied cytotoxic T lymphocytes (CTLs) in which expression of the T cell receptor–activated tyrosine kinase Lck is ablated. In the absence of Lck, the centrosome is able to translocate around the nucleus toward the immunological synapse but is unable to dock at the plasma membrane. Lytic granules fail to polarize and release their contents, and target cells are not killed. In CTLs deficient in both Lck and the related tyrosine kinase Fyn, centrosome translocation is impaired, and the centrosome remains on the distal side of the nucleus relative to the synapse. These results show that repositioning of the centrosome in CTLs involves at least two distinct steps, with Lck signaling required for the centrosome to dock at the plasma membrane. PMID:21339332

Developmental alterations in centrosome integrity contribute to the post-mitotic state of mammalian cardiomyocytes

PubMed Central

Zebrowski, David C; Vergarajauregui, Silvia; Wu, Chi-Chung; Piatkowski, Tanja; Becker, Robert; Leone, Marina; Hirth, Sofia; Ricciardi, Filomena; Falk, Nathalie; Giessl, Andreas; Just, Steffen; Braun, Thomas; Weidinger, Gilbert; Engel, Felix B

2015-01-01

Mammalian cardiomyocytes become post-mitotic shortly after birth. Understanding how this occurs is highly relevant to cardiac regenerative therapy. Yet, how cardiomyocytes achieve and maintain a post-mitotic state is unknown. Here, we show that cardiomyocyte centrosome integrity is lost shortly after birth. This is coupled with relocalization of various centrosome proteins to the nuclear envelope. Consequently, postnatal cardiomyocytes are unable to undergo ciliogenesis and the nuclear envelope adopts the function as cellular microtubule organizing center. Loss of centrosome integrity is associated with, and can promote, cardiomyocyte G0/G1 cell cycle arrest suggesting that centrosome disassembly is developmentally utilized to achieve the post-mitotic state in mammalian cardiomyocytes. Adult cardiomyocytes of zebrafish and newt, which are able to proliferate, maintain centrosome integrity. Collectively, our data provide a novel mechanism underlying the post-mitotic state of mammalian cardiomyocytes as well as a potential explanation for why zebrafish and newts, but not mammals, can regenerate their heart. DOI: http://dx.doi.org/10.7554/eLife.05563.001 PMID:26247711

VDAC3 regulates centriole assembly by targeting Mps1 to centrosomes.

PubMed

Majumder, Shubhra; Slabodnick, Mark; Pike, Amanda; Marquardt, Joseph; Fisk, Harold A

2012-10-01

Centrioles are duplicated during S-phase to generate the two centrosomes that serve as mitotic spindle poles during mitosis. The centrosomal pool of the Mps1 kinase is important for centriole assembly, but how Mps1 is delivered to centrosomes is unknown. Here we have identified a centrosome localization domain within Mps1 and identified the mitochondrial porin VDAC3 as a protein that binds to this region of Mps1. Moreover, we show that VDAC3 is present at the mother centriole and modulates centriole assembly by recruiting Mps1 to centrosomes.

VDAC3 regulates centriole assembly by targeting Mps1 to centrosomes

PubMed Central

Majumder, Shubhra; Slabodnick, Mark; Pike, Amanda; Marquardt, Joseph; Fisk, Harold A.

2012-01-01

Centrioles are duplicated during S-phase to generate the two centrosomes that serve as mitotic spindle poles during mitosis. The centrosomal pool of the Mps1 kinase is important for centriole assembly, but how Mps1 is delivered to centrosomes is unknown. Here we have identified a centrosome localization domain within Mps1 and identified the mitochondrial porin VDAC3 as a protein that binds to this region of Mps1. Moreover, we show that VDAC3 is present at the mother centriole and modulates centriole assembly by recruiting Mps1 to centrosomes. PMID:22935710

Germ Cell-less Promotes Centrosome Segregation to Induce Germ Cell Formation.

PubMed

Lerit, Dorothy A; Shebelut, Conrad W; Lawlor, Kristen J; Rusan, Nasser M; Gavis, Elizabeth R; Schedl, Paul; Deshpande, Girish

2017-01-24

The primordial germ cells (PGCs) specified during embryogenesis serve as progenitors to the adult germline stem cells. In Drosophila, the proper specification and formation of PGCs require both centrosomes and germ plasm, which contains the germline determinants. Centrosomes are microtubule (MT)-organizing centers that ensure the faithful segregation of germ plasm into PGCs. To date, mechanisms that modulate centrosome behavior to engineer PGC development have remained elusive. Only one germ plasm component, Germ cell-less (Gcl), is known to play a role in PGC formation. Here, we show that Gcl engineers PGC formation by regulating centrosome dynamics. Loss of gcl leads to aberrant centrosome separation and elaboration of the astral MT network, resulting in inefficient germ plasm segregation and aborted PGC cellularization. Importantly, compromising centrosome separation alone is sufficient to mimic the gcl loss-of-function phenotypes. We conclude Gcl functions as a key regulator of centrosome separation required for proper PGC development. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

EGF Induced Centrosome Separation Promotes Mitotic Progression and Cell Survival

PubMed Central

Mardin, Balca R.; Isokane, Mayumi; Cosenza, Marco R.; Krämer, Alwin; Ellenberg, Jan; Fry, Andrew M.; Schiebel, Elmar

2014-01-01

Summary Timely and accurate assembly of the mitotic spindle is critical for the faithful segregation of chromosomes and centrosome separation is a key step in this process. The timing of centrosome separation varies dramatically between cell types; however, the mechanisms responsible for these differences and its significance are unclear. Here, we show that activation of epidermal growth factor receptor (EGFR) signaling determines the timing of centrosome separation. Premature separation of centrosomes decreases the requirement for the major mitotic kinesin Eg5 for spindle assembly, accelerates mitosis and decreases the rate of chromosome missegregation. Importantly, EGF stimulation impacts upon centrosome separation and mitotic progression to different degrees in different cell lines. Cells with high EGFR levels fail to arrest in mitosis upon Eg5 inhibition. This has important implications for cancer therapy since cells with high centrosomal response to EGF are more susceptible to combinatorial inhibition of EGFR and Eg5. PMID:23643362

Natural product-inspired cascade synthesis yields modulators of centrosome integrity.

PubMed

Dückert, Heiko; Pries, Verena; Khedkar, Vivek; Menninger, Sascha; Bruss, Hanna; Bird, Alexander W; Maliga, Zoltan; Brockmeyer, Andreas; Janning, Petra; Hyman, Anthony; Grimme, Stefan; Schürmann, Markus; Preut, Hans; Hübel, Katja; Ziegler, Slava; Kumar, Kamal; Waldmann, Herbert

2011-12-25

In biology-oriented synthesis, the scaffolds of biologically relevant compound classes inspire the synthesis of focused compound collections enriched in bioactivity. This criterion is, in particular, met by the scaffolds of natural products selected in evolution. The synthesis of natural product-inspired compound collections calls for efficient reaction sequences that preferably combine multiple individual transformations in one operation. Here we report the development of a one-pot, twelve-step cascade reaction sequence that includes nine different reactions and two opposing kinds of organocatalysis. The cascade sequence proceeds within 10-30 min and transforms readily available substrates into complex indoloquinolizines that resemble the core tetracyclic scaffold of numerous polycyclic indole alkaloids. Biological investigation of a corresponding focused compound collection revealed modulators of centrosome integrity, termed centrocountins, which caused fragmented and supernumerary centrosomes, chromosome congression defects, multipolar mitotic spindles, acentrosomal spindle poles and multipolar cell division by targeting the centrosome-associated proteins nucleophosmin and Crm1.

Centrosome-Based Mechanisms, Prognostics and Therapeutics in Prostate Cancer

DTIC Science & Technology

2006-12-01

progression of prostate carcinomas. The specific aims of the original proposal were designed to test several features of this model . 1. Are centrosome...features of this model . 1. Are centrosome defects present in early prostate cancer and can they predict aggressive disease? 2. Do pericentrin’s...cells, supports this model . The ability to block the cell cycle in prostate cells by depletion of any of 14 centrosome proteins identifies several

Centrioles, centrosomes, and cilia in health and disease.

PubMed

Nigg, Erich A; Raff, Jordan W

2009-11-13

Centrioles are barrel-shaped structures that are essential for the formation of centrosomes, cilia, and flagella. Here we review recent advances in our understanding of the function and biogenesis of these organelles, and we emphasize their connection to human disease. Deregulation of centrosome numbers has long been proposed to contribute to genome instability and tumor formation, whereas mutations in centrosomal proteins have recently been genetically linked to microcephaly and dwarfism. Finally, structural or functional centriole aberrations contribute to ciliopathies, a variety of complex diseases that stem from the absence or dysfunction of cilia.

Stabilization of cartwheel-less centrioles for duplication requires CEP295-mediated centriole-to-centrosome conversion.

PubMed

Izquierdo, Denisse; Wang, Won-Jing; Uryu, Kunihiro; Tsou, Meng-Fu Bryan

2014-08-21

Vertebrate centrioles lose their geometric scaffold, the cartwheel, during mitosis, concurrently with gaining the ability to recruit the pericentriolar material (PCM) and thereby function as the centrosome. Cartwheel removal has recently been implicated in centriole duplication, but whether "cartwheel-less" centrioles are intrinsically stable or must be maintained through other modifications remains unclear. Here, we identify a newborn centriole-enriched protein, KIAA1731/CEP295, specifically mediating centriole-to-centrosome conversion but dispensable for cartwheel removal. In the absence of CEP295, centrioles form in the S/G2 phase and lose their associated cartwheel in mitosis but cannot be converted to centrosomes, uncoupling the two events. Strikingly, centrioles devoid of both the PCM and the cartwheel progressively lose centriolar components, whereas centrioles associating with either the cartwheel or PCM alone can exist stably. Thus, cartwheel removal can have grave repercussions to centriole stability, and centriole-to-centrosome conversion mediated by CEP295 must occur in parallel to maintain cartwheel-less centrioles for duplication. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

E2F Activators Signal and Maintain Centrosome Amplification in Breast Cancer Cells

PubMed Central

Lee, Mi-Young; Moreno, Carlos S.

2014-01-01

Centrosomes ensure accurate chromosome segregation by directing spindle bipolarity. Loss of centrosome regulation results in centrosome amplification, multipolar mitosis and aneuploidy. Since centrosome amplification is common in premalignant lesions and breast tumors, it is proposed to play a central role in breast tumorigenesis, a hypothesis that remains to be tested. The coordination between the cell and centrosome cycles is of paramount importance to maintain normal centrosome numbers, and the E2Fs may be responsible for regulating these cycles. However, the role of E2F activators in centrosome amplification is unclear. Because E2Fs are deregulated in Her2+ cells displaying centrosome amplification, we addressed whether they signal this abnormal process. Knockdown of E2F1 or E2F3 in Her2+ cells decreased centrosome amplification without significantly affecting cell cycle progression, whereas the overexpression of E2F1, E2F2, or E2F3 increased centrosome amplification in MCF10A mammary epithelial cells. Our results revealed that E2Fs affect the expression of proteins, including Nek2 and Plk4, known to influence the cell/centrosome cycles and mitosis. Downregulation of E2F3 resulted in cell death and delays/blocks in cytokinesis, which was reversed by Nek2 overexpression. Nek2 overexpression enhanced centrosome amplification in Her2+ breast cancer cells silenced for E2F3, revealing a role for the E2F activators in maintaining centrosome amplification in part through Nek2. PMID:24797070

E2F activators signal and maintain centrosome amplification in breast cancer cells.

PubMed

Lee, Mi-Young; Moreno, Carlos S; Saavedra, Harold I

2014-07-01

Centrosomes ensure accurate chromosome segregation by directing spindle bipolarity. Loss of centrosome regulation results in centrosome amplification, multipolar mitosis and aneuploidy. Since centrosome amplification is common in premalignant lesions and breast tumors, it is proposed to play a central role in breast tumorigenesis, a hypothesis that remains to be tested. The coordination between the cell and centrosome cycles is of paramount importance to maintain normal centrosome numbers, and the E2Fs may be responsible for regulating these cycles. However, the role of E2F activators in centrosome amplification is unclear. Because E2Fs are deregulated in Her2(+) cells displaying centrosome amplification, we addressed whether they signal this abnormal process. Knockdown of E2F1 or E2F3 in Her2(+) cells decreased centrosome amplification without significantly affecting cell cycle progression, whereas the overexpression of E2F1, E2F2, or E2F3 increased centrosome amplification in MCF10A mammary epithelial cells. Our results revealed that E2Fs affect the expression of proteins, including Nek2 and Plk4, known to influence the cell/centrosome cycles and mitosis. Downregulation of E2F3 resulted in cell death and delays/blocks in cytokinesis, which was reversed by Nek2 overexpression. Nek2 overexpression enhanced centrosome amplification in Her2(+) breast cancer cells silenced for E2F3, revealing a role for the E2F activators in maintaining centrosome amplification in part through Nek2.

Disruption of clathrin-mediated trafficking causes centrosome overduplication and senescence.

PubMed

Olszewski, Maciej B; Chandris, Panagiotis; Park, Bum-Chan; Eisenberg, Evan; Greene, Lois E

2014-01-01

The Hsc70 cochaperone, G cyclin-associated kinase (GAK), has been shown to be essential for the chaperoning of clathrin by Hsc70 in the cell. In this study, we used conditional GAK knockout mouse embryonic fibroblasts (MEFs) to determine the effect of completely inhibiting clathrin-dependent trafficking on the cell cycle. After GAK was knocked out, the cells developed the unusual phenotype of having multiple centrosomes, but at the same time failed to divide and ultimately became senescent. To explain this phenotype, we examined the signaling profile and found that mitogenic stimulation of the GAK KO cells and the control cells were similar except for increased phosphorylation of Akt. In addition, the disruption of intracellular trafficking caused by knocking out GAK destabilized the lysosomal membranes, resulting in DNA damage due to iron leakage. Knocking down clathrin heavy chain or inhibiting dynamin largely reproduced the GAK KO phenotype, but inhibiting only clathrin-mediated endocytosis by knocking down adaptor protein (AP2) caused growth arrest and centrosome overduplication, but no DNA damage or senescence. We conclude that disruption of clathrin-dependent trafficking induces senescence accompanied by centrosome overduplication because of a combination of DNA damage and changes in mitogenic signaling that uncouples centrosomal duplication from DNA replication. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

Centriolar satellites assemble centrosomal microcephaly proteins to recruit CDK2 and promote centriole duplication

PubMed Central

Kodani, Andrew; Yu, Timothy W; Johnson, Jeffrey R; Jayaraman, Divya; Johnson, Tasha L; Al-Gazali, Lihadh; Sztriha, Lāszló; Partlow, Jennifer N; Kim, Hanjun; Krup, Alexis L; Dammermann, Alexander; Krogan, Nevan J; Walsh, Christopher A; Reiter, Jeremy F

2015-01-01

Primary microcephaly (MCPH) associated proteins CDK5RAP2, CEP152, WDR62 and CEP63 colocalize at the centrosome. We found that they interact to promote centriole duplication and form a hierarchy in which each is required to localize another to the centrosome, with CDK5RAP2 at the apex, and CEP152, WDR62 and CEP63 at sequentially lower positions. MCPH proteins interact with distinct centriolar satellite proteins; CDK5RAP2 interacts with SPAG5 and CEP72, CEP152 with CEP131, WDR62 with MOONRAKER, and CEP63 with CEP90 and CCDC14. These satellite proteins localize their cognate MCPH interactors to centrosomes and also promote centriole duplication. Consistent with a role for satellites in microcephaly, homozygous mutations in one satellite gene, CEP90, may cause MCPH. The satellite proteins, with the exception of CCDC14, and MCPH proteins promote centriole duplication by recruiting CDK2 to the centrosome. Thus, centriolar satellites build a MCPH complex critical for human neurodevelopment that promotes CDK2 centrosomal localization and centriole duplication. DOI: http://dx.doi.org/10.7554/eLife.07519.001 PMID:26297806

Phosphorylation of DEPDC1 at Ser110 is required to maintain centrosome organization during mitosis.

PubMed

Chen, Dan; Ito, Satoko; Hyodo, Toshinori; Asano-Inami, Eri; Yuan, Hong; Senga, Takeshi

2017-09-15

DEPDC1 (DEP domain containing 1) is overexpressed in multiple cancers and is associated with cell cycle progression. In this report, we have investigated the expression, localization, phosphorylation and function of DEPDC1 during mitosis. DEPDC1 has two isoforms (isoform a and isoform b), and both of them are increased in mitosis and degraded once cells exit mitosis. DEPDC1a is localized to the centrosome in metaphase, whereas DEPDC1b is localized to the entire cell cortex during mitosis. DEPDC1a, but not DEPDC1b, was required for the integrity of centrosome and organization of the bipolar spindle. Mass spectrometry and biochemical analyses revealed phosphorylation of DEPDC1 at Ser110. The phosphorylation of Ser110 is essential for localization of DEPDC1a to the centrosome. Consistently, non-phosphorylation mutants of DEPDC1a did not rescue disruption of centrosome organization by depletion of endogenous DEPDC1. Our results show a novel role for DEPDC1 in maintaining centrosome integrity during mitosis for the accurate distribution of chromosomes. Copyright © 2017. Published by Elsevier Inc.

Coiled-Coil Proteins Facilitated the Functional Expansion of the Centrosome

PubMed Central

Kuhn, Michael; Hyman, Anthony A.; Beyer, Andreas

2014-01-01

Repurposing existing proteins for new cellular functions is recognized as a main mechanism of evolutionary innovation, but its role in organelle evolution is unclear. Here, we explore the mechanisms that led to the evolution of the centrosome, an ancestral eukaryotic organelle that expanded its functional repertoire through the course of evolution. We developed a refined sequence alignment technique that is more sensitive to coiled coil proteins, which are abundant in the centrosome. For proteins with high coiled-coil content, our algorithm identified 17% more reciprocal best hits than BLAST. Analyzing 108 eukaryotic genomes, we traced the evolutionary history of centrosome proteins. In order to assess how these proteins formed the centrosome and adopted new functions, we computationally emulated evolution by iteratively removing the most recently evolved proteins from the centrosomal protein interaction network. Coiled-coil proteins that first appeared in the animal–fungi ancestor act as scaffolds and recruit ancestral eukaryotic proteins such as kinases and phosphatases to the centrosome. This process created a signaling hub that is crucial for multicellular development. Our results demonstrate how ancient proteins can be co-opted to different cellular localizations, thereby becoming involved in novel functions. PMID:24901223

Design, Synthesis, and Biological Evaluation of an Allosteric Inhibitor of HSET that Targets Cancer Cells with Supernumerary Centrosomes

PubMed Central

Watts, Ciorsdaidh A.; Richards, Frances M.; Bender, Andreas; Bond, Peter J.; Korb, Oliver; Kern, Oliver; Riddick, Michelle; Owen, Paul; Myers, Rebecca M.; Raff, Jordan; Gergely, Fanni; Jodrell, Duncan I.; Ley, Steven V.

2013-01-01

Summary Centrosomes associate with spindle poles; thus, the presence of two centrosomes promotes bipolar spindle assembly in normal cells. Cancer cells often contain supernumerary centrosomes, and to avoid multipolar mitosis and cell death, these are clustered into two poles by the microtubule motor protein HSET. We report the discovery of an allosteric inhibitor of HSET, CW069, which we designed using a methodology on an interface of chemistry and biology. Using this approach, we explored millions of compounds in silico and utilized convergent syntheses. Only compound CW069 showed marked activity against HSET in vitro. The inhibitor induced multipolar mitoses only in cells containing supernumerary centrosomes. CW069 therefore constitutes a valuable tool for probing HSET function and, by reducing the growth of cells containing supernumerary centrosomes, paves the way for new cancer therapeutics. PMID:24210220

Structural protein 4.1 is located in mammalian centrosomes

PubMed Central

Krauss, Sharon Wald; Chasis, Joel Anne; Rogers, Catherine; Mohandas, Narla; Krockmalnic, Gabriela; Penman, Sheldon

1997-01-01

Structural protein 4.1 was first characterized as an important 80-kDa protein in the mature red cell membrane skeleton. It is now known to be a member of a family of protein isoforms detected at diverse intracellular sites in many nucleated mammalian cells. We recently reported that protein 4.1 isoforms are present at interphase in nuclear matrix and are rearranged during the cell cycle. Here we report that protein 4.1 epitopes are present in centrosomes of human and murine cells and are detected by using affinity-purified antibodies specific for 80-kDa red cell 4.1 and for 4.1 peptides. Immunofluorescence, by both conventional and confocal microscopy, showed that protein 4.1 epitopes localized in the pericentriolar region. Protein 4.1 epitopes remained in centrosomes after extraction of cells with detergent, salt, and DNase. Higher resolution electron microscopy of detergent-extracted cell whole mounts showed centrosomal protein 4.1 epitopes distributed along centriolar cylinders and on pericentriolar fibers, at least some of which constitute the filamentous network surrounding each centriole. Double-label electron microscopy showed that protein 4.1 epitopes were predominately localized in regions also occupied by epitopes for centrosome-specific autoimmune serum 5051 but were not found on microtubules. Our results suggest that protein 4.1 is an integral component of centrosome structure, in which it may play an important role in centrosome function during cell division and organization of cellular architecture. PMID:9207085

Structural centrosome aberrations sensitize polarized epithelia to basal cell extrusion.

PubMed

Ganier, Olivier; Schnerch, Dominik; Nigg, Erich A

2018-06-01

Centrosome aberrations disrupt tissue architecture and may confer invasive properties to cancer cells. Here we show that structural centrosome aberrations, induced by overexpression of either Ninein-like protein (NLP) or CEP131/AZI1, sensitize polarized mammalian epithelia to basal cell extrusion. While unperturbed epithelia typically dispose of damaged cells through apical dissemination into luminal cavities, certain oncogenic mutations cause a switch in directionality towards basal cell extrusion, raising the potential for metastatic cell dissemination. Here we report that NLP-induced centrosome aberrations trigger the preferential extrusion of damaged cells towards the basal surface of epithelial monolayers. This switch in directionality from apical to basal dissemination coincides with a profound reorganization of the microtubule cytoskeleton, which in turn prevents the contractile ring repositioning that is required to support extrusion towards the apical surface. While the basal extrusion of cells harbouring NLP-induced centrosome aberrations requires exogenously induced cell damage, structural centrosome aberrations induced by excess CEP131 trigger the spontaneous dissemination of dying cells towards the basal surface from MDCK cysts. Thus, similar to oncogenic mutations, structural centrosome aberrations can favour basal extrusion of damaged cells from polarized epithelia. Assuming that additional mutations may promote cell survival, this process could sensitize epithelia to disseminate potentially metastatic cells. © 2018 The Authors.

Conserved molecular interactions in centriole-to-centrosome conversion.

PubMed

Fu, Jingyan; Lipinszki, Zoltan; Rangone, Hélène; Min, Mingwei; Mykura, Charlotte; Chao-Chu, Jennifer; Schneider, Sandra; Dzhindzhev, Nikola S; Gottardo, Marco; Riparbelli, Maria Giovanna; Callaini, Giuliano; Glover, David M

2016-01-01

Centrioles are required to assemble centrosomes for cell division and cilia for motility and signalling. New centrioles assemble perpendicularly to pre-existing ones in G1-S and elongate throughout S and G2. Fully elongated daughter centrioles are converted into centrosomes during mitosis to be able to duplicate and organize pericentriolar material in the next cell cycle. Here we show that centriole-to-centrosome conversion requires sequential loading of Cep135, Ana1 (Cep295) and Asterless (Cep152) onto daughter centrioles during mitotic progression in both Drosophila melanogaster and human. This generates a molecular network spanning from the inner- to outermost parts of the centriole. Ana1 forms a molecular strut within the network, and its essential role can be substituted by an engineered fragment providing an alternative linkage between Asterless and Cep135. This conserved architectural framework is essential for loading Asterless or Cep152, the partner of the master regulator of centriole duplication, Plk4. Our study thus uncovers the molecular basis for centriole-to-centrosome conversion that renders daughter centrioles competent for motherhood.

The daughter centriole controls ciliogenesis by regulating Neurl-4 localization at the centrosome

PubMed Central

Loukil, Abdelhalim; Tormanen, Kati

2017-01-01

The two centrioles of the centrosome differ in age and function. Although the mother centriole mediates most centrosome-dependent processes, the role of the daughter remains poorly understood. A recent study has implicated the daughter centriole in centriole amplification in multiciliated cells, but its contribution to primary ciliogenesis is unclear. We found that manipulations that prevent daughter centriole formation or induce its separation from the mother abolish ciliogenesis. This defect was caused by stabilization of the negative ciliogenesis regulator CP110 and was corrected by CP110 depletion. CP110 dysregulation may be caused by effects on Neurl-4, a daughter centriole–associated ubiquitin ligase cofactor, which was required for ciliogenesis. Centrosome-targeted Neurl-4 was sufficient to restore ciliogenesis in cells with manipulated daughter centrioles. Interestingly, early during ciliogenesis, Neurl-4 transiently associated with the mother centriole in a process that required mother–daughter centriole proximity. Our data support a model in which the daughter centriole promotes ciliogenesis through Neurl-4–dependent regulation of CP110 levels at the mother centriole. PMID:28385950

An intact centrosome is required for the maintenance of polarization during directional cell migration.

PubMed

Wakida, Nicole M; Botvinick, Elliot L; Lin, Justin; Berns, Michael W

2010-12-23

Establishing and maintaining polarization is critical during cell migration. It is known that the centrosome contains numerous proteins whose roles of organizing the microtubule network range include nucleation, stabilization and severing. It is not known whether the centrosome is necessary to maintain polarization. Due to its role as the microtubule organizing center, we hypothesize that the centrosome is necessary to maintain polarization in a migrating cell. Although there have been implications of its role in cell migration, there is no direct study of the centrosome's role in maintaining polarization. In this study we ablate the centrosome by intracellular laser irradiation to understand the role of the centrosome in two vastly different cell types, human osteosarcoma (U2OS) and rat kangaroo kidney epithelial cells (PtK). The PtK cell line has been extensively used as a model for cytoskeletal dynamics during cell migration. The U2OS cell line serves as a model for a complex, single migrating cell. In this study we use femtosecond near-infrared laser irradiation to remove the centrosome in migrating U2OS and PtK2 cells. Immunofluorescence staining for centrosomal markers verified successful irradiation with 94% success. A loss of cell polarization is observed between 30 and 90 minutes following removal of the centrosome. Changes in cell shape are correlated with modifications in microtubule and actin organization. Changes in cell morphology and microtubule organization were quantified revealing significant depolarization resulting from centrosome irradiation. This study demonstrates that the centrosome is necessary for the maintenance of polarization during directed cell migration in two widely different cell types. Removal of the centrosome from a polarized cell results in the reorganization of the microtubule network into a symmetric non-polarized phenotype. These results demonstrate that the centrosome plays a critical role in the maintenance of cytoskeletal

APC functions at the centrosome to stimulate microtubule growth.

PubMed

Lui, Christina; Ashton, Cahora; Sharma, Manisha; Brocardo, Mariana G; Henderson, Beric R

2016-01-01

The adenomatous polyposis coli (APC) tumor suppressor is multi-functional. APC is known to localize at the centrosome, and in mitotic cells contributes to formation of the mitotic spindle. To test whether APC contributes to nascent microtubule (MT) growth at interphase centrosomes, we employed MT regrowth assays in U2OS cells to measure MT assembly before and after nocodazole treatment and release. We showed that siRNA knockdown of full-length APC delayed both initial MT aster formation and MT elongation/regrowth. In contrast, APC-mutant SW480 cancer cells displayed a defect in MT regrowth that was unaffected by APC knockdown, but which was rescued by reconstitution of full-length APC. Our findings identify APC as a positive regulator of centrosome MT initial assembly and suggest that this process is disrupted by cancer mutations. We confirmed that full-length APC associates with the MT-nucleation factor γ-tubulin, and found that the APC cancer-truncated form (1-1309) also bound to γ-tubulin through APC amino acids 1-453. While binding to γ-tubulin may help target APC to the site of MT nucleation complexes, additional C-terminal sequences of APC are required to stimulate and stabilize MT growth. Copyright © 2015 Elsevier Ltd. All rights reserved.

Control of endothelial cell polarity and sprouting angiogenesis by non-centrosomal microtubules

PubMed Central

Martin, Maud; Veloso, Alexandra; Wu, Jingchao; Katrukha, Eugene A

2018-01-01

Microtubules control different aspects of cell polarization. In cells with a radial microtubule system, a pivotal role in setting up asymmetry is attributed to the relative positioning of the centrosome and the nucleus. Here, we show that centrosome loss had no effect on the ability of endothelial cells to polarize and move in 2D and 3D environments. In contrast, non-centrosomal microtubules stabilized by the microtubule minus-end-binding protein CAMSAP2 were required for directional migration on 2D substrates and for the establishment of polarized cell morphology in soft 3D matrices. CAMSAP2 was also important for persistent endothelial cell sprouting during in vivo zebrafish vessel development. In the absence of CAMSAP2, cell polarization in 3D could be partly rescued by centrosome depletion, indicating that in these conditions the centrosome inhibited cell polarity. We propose that CAMSAP2-protected non-centrosomal microtubules are needed for establishing cell asymmetry by enabling microtubule enrichment in a single-cell protrusion. PMID:29547120

The Centrioles, Centrosomes, Basal Bodies, and Cilia of Drosophila melanogaster

PubMed Central

Lattao, Ramona; Kovács, Levente; Glover, David M.

2017-01-01

Centrioles play a key role in the development of the fly. They are needed for the correct formation of centrosomes, the organelles at the poles of the spindle that can persist as microtubule organizing centers (MTOCs) into interphase. The ability to nucleate cytoplasmic microtubules (MTs) is a property of the surrounding pericentriolar material (PCM). The centriole has a dual life, existing not only as the core of the centrosome but also as the basal body, the structure that templates the formation of cilia and flagellae. Thus the structure and functions of the centriole, the centrosome, and the basal body have an impact upon many aspects of development and physiology that can readily be modeled in Drosophila. Centrosomes are essential to give organization to the rapidly increasing numbers of nuclei in the syncytial embryo and for the spatially precise execution of cell division in numerous tissues, particularly during male meiosis. Although mitotic cell cycles can take place in the absence of centrosomes, this is an error-prone process that opens up the fly to developmental defects and the potential of tumor formation. Here, we review the structure and functions of the centriole, the centrosome, and the basal body in different tissues and cultured cells of Drosophila melanogaster, highlighting their contributions to different aspects of development and cell division. PMID:28476861

The Centrioles, Centrosomes, Basal Bodies, and Cilia of Drosophila melanogaster.

PubMed

Lattao, Ramona; Kovács, Levente; Glover, David M

2017-05-01

Centrioles play a key role in the development of the fly. They are needed for the correct formation of centrosomes, the organelles at the poles of the spindle that can persist as microtubule organizing centers (MTOCs) into interphase. The ability to nucleate cytoplasmic microtubules (MTs) is a property of the surrounding pericentriolar material (PCM). The centriole has a dual life, existing not only as the core of the centrosome but also as the basal body, the structure that templates the formation of cilia and flagellae. Thus the structure and functions of the centriole, the centrosome, and the basal body have an impact upon many aspects of development and physiology that can readily be modeled in Drosophila Centrosomes are essential to give organization to the rapidly increasing numbers of nuclei in the syncytial embryo and for the spatially precise execution of cell division in numerous tissues, particularly during male meiosis. Although mitotic cell cycles can take place in the absence of centrosomes, this is an error-prone process that opens up the fly to developmental defects and the potential of tumor formation. Here, we review the structure and functions of the centriole, the centrosome, and the basal body in different tissues and cultured cells of Drosophila melanogaster , highlighting their contributions to different aspects of development and cell division. Copyright © 2017 Lattao et al.

Mother Centriole Distal Appendages Mediate Centrosome Docking at the Immunological Synapse and Reveal Mechanistic Parallels with Ciliogenesis.

PubMed

Stinchcombe, Jane C; Randzavola, Lyra O; Angus, Karen L; Mantell, Judith M; Verkade, Paul; Griffiths, Gillian M

2015-12-21

Cytotoxic T lymphocytes (CTLs) are highly effective serial killers capable of destroying virally infected and cancerous targets by polarized release from secretory lysosomes. Upon target contact, the CTL centrosome rapidly moves to the immunological synapse, focusing microtubule-directed release at this point [1-3]. Striking similarities have been noted between centrosome polarization at the synapse and basal body docking during ciliogenesis [1, 4-8], suggesting that CTL centrosomes might dock with the plasma membrane during killing, in a manner analogous to primary cilia formation [1, 4]. However, questions remain regarding the extent and function of centrosome polarization at the synapse, and recent reports have challenged its role [9, 10]. Here, we use high-resolution transmission electron microscopy (TEM) tomography analysis to show that, as in ciliogenesis, the distal appendages of the CTL mother centriole contact the plasma membrane directly during synapse formation. This is functionally important as small interfering RNA (siRNA) targeting of the distal appendage protein, Cep83, required for membrane contact during ciliogenesis [11], impairs CTL secretion. Furthermore, the regulatory proteins CP110 and Cep97, which must dissociate from the mother centriole to allow cilia formation [12], remain associated with the mother centriole in CTLs, and neither axoneme nor transition zone ciliary structures form. Moreover, complete centrosome docking can occur in proliferating CTLs with multiple centriole pairs. Thus, in CTLs, centrosomes dock transiently with the membrane, within the cell cycle and without progression into ciliogenesis. We propose that this transient centrosome docking without cilia formation is important for CTLs to deliver rapid, repeated polarized secretion directed by the centrosome. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Microtubule release from the centrosome in migrating cells

PubMed Central

Abal, Miguel; Piel, Matthieu; Bouckson-Castaing, Veronique; Mogensen, Mette; Sibarita, Jean-Baptiste; Bornens, Michel

2002-01-01

In migrating cells, force production relies essentially on a polarized actomyosin system, whereas the spatial regulation of actomyosin contraction and substrate contact turnover involves a complex cooperation between the microtubule (MT) and the actin filament networks (Goode, B.L., D.G. Drubin, and G. Barnes. 2000. Curr. Opin. Cell Biol., 12:63–71). Targeting and capture of MT plus ends at the cell periphery has been described, but whether or not the minus ends of these MTs are anchored at the centrosome is not known. Here, we show that release of short MTs from the centrosome is frequent in migrating cells and that their transport toward the cell periphery is blocked when dynein activity is impaired. We further show that MT release, but not MT nucleation or polymerization dynamics, is abolished by overexpression of the centrosomal MT-anchoring protein ninein. In addition, a dramatic inhibition of cell migration was observed; but, contrary to cells treated by drugs inhibiting MT dynamics, polarized membrane ruffling activity was not affected in ninein overexpressing cells. We thus propose that the balance between MT minus-end capture and release from the centrosome is critical for efficient cell migration. PMID:12473683

Phosphorylation of Nlp by Plk1 negatively regulates its dynein-dynactin-dependent targeting to the centrosome.

PubMed

Casenghi, Martina; Barr, Francis A; Nigg, Erich A

2005-11-01

When cells enter mitosis the microtubule (MT) network undergoes a profound rearrangement, in part due to alterations in the MT nucleating and anchoring properties of the centrosome. Ninein and the ninein-like protein (Nlp) are centrosomal proteins involved in MT organisation in interphase cells. We show that the overexpression of these two proteins induces the fragmentation of the Golgi, and causes lysosomes to disperse toward the cell periphery. The ability of Nlp and ninein to perturb the cytoplasmic distribution of these organelles depends on their ability to interact with the dynein-dynactin motor complex. Our data also indicate that dynactin is required for the targeting of Nlp and ninein to the centrosome. Furthermore, phosphorylation of Nlp by the polo-like kinase 1 (Plk1) negatively regulates its association with dynactin. These findings uncover a mechanism through which Plk1 helps to coordinate changes in MT organisation with cell cycle progression, by controlling the dynein-dynactin-dependent transport of centrosomal proteins.

Behavior of centrosomes during fertilization and cell division in mouse oocytes and in sea urchin eggs

NASA Technical Reports Server (NTRS)

Schatten, Heide; Schatten, Gerald; Balczon, Ron; Simerly, Calvin; Mazia, Daniel

1986-01-01

The behavior of centrosomes during the stages of fertilization and cell division in mouse oocytes and in sea urchin eggs was monitored in an immunofluorescence microscope, using autoimmune centrosomal antiserum derived from a patient with scleroderma to label the centrosomal material. These observations showed that centrosomes reproduce during the interphase and aggregate and separate during cell mitosis. Results supported the hypothesis of Mazia (1984), who proposed that centrosomes are 'flexible bodies'. It was also found that, while the sea urchin centrosomes are paternally inherited as was initially proposed by Bovery (1904), the mouse centrosomes are of maternal origin.

Drosophila Ana1 is required for centrosome assembly and centriole elongation.

PubMed

Saurya, Saroj; Roque, Hélio; Novak, Zsofia A; Wainman, Alan; Aydogan, Mustafa G; Volanakis, Adam; Sieber, Boris; Pinto, David Miguel Susano; Raff, Jordan W

2016-07-01

Centrioles organise centrosomes and cilia, and these organelles have an important role in many cell processes. In flies, the centriole protein Ana1 is required for the assembly of functional centrosomes and cilia. It has recently been shown that Cep135 (also known as Bld10) initially recruits Ana1 to newly formed centrioles, and that Ana1 then recruits Asl (known as Cep152 in mammals) to promote the conversion of these centrioles into centrosomes. Here, we show that ana1 mutants lack detectable centrosomes in vivo, that Ana1 is irreversibly incorporated into centrioles during their assembly and appears to play a more important role in maintaining Asl at centrioles than in initially recruiting Asl to centrioles. Unexpectedly, we also find that Ana1 promotes centriole elongation in a dose-dependent manner: centrioles are shorter when Ana1 dosage is reduced and are longer when Ana1 is overexpressed. This latter function of Ana1 appears to be distinct from its role in centrosome and cilium function, as a GFP-Ana1 fusion lacking the N-terminal 639 amino acids of the protein can support centrosome assembly and cilium function but cannot promote centriole over-elongation when overexpressed. © 2016. Published by The Company of Biologists Ltd.

Drosophila Ana1 is required for centrosome assembly and centriole elongation

PubMed Central

Saurya, Saroj; Roque, Hélio; Novak, Zsofia A.; Wainman, Alan; Aydogan, Mustafa G.; Volanakis, Adam; Sieber, Boris; Pinto, David Miguel Susano

2016-01-01

ABSTRACT Centrioles organise centrosomes and cilia, and these organelles have an important role in many cell processes. In flies, the centriole protein Ana1 is required for the assembly of functional centrosomes and cilia. It has recently been shown that Cep135 (also known as Bld10) initially recruits Ana1 to newly formed centrioles, and that Ana1 then recruits Asl (known as Cep152 in mammals) to promote the conversion of these centrioles into centrosomes. Here, we show that ana1 mutants lack detectable centrosomes in vivo, that Ana1 is irreversibly incorporated into centrioles during their assembly and appears to play a more important role in maintaining Asl at centrioles than in initially recruiting Asl to centrioles. Unexpectedly, we also find that Ana1 promotes centriole elongation in a dose-dependent manner: centrioles are shorter when Ana1 dosage is reduced and are longer when Ana1 is overexpressed. This latter function of Ana1 appears to be distinct from its role in centrosome and cilium function, as a GFP–Ana1 fusion lacking the N-terminal 639 amino acids of the protein can support centrosome assembly and cilium function but cannot promote centriole over-elongation when overexpressed. PMID:27206860

Centrobin-Centrosomal Protein 4.1-associated Protein (CPAP) Interaction Promotes CPAP Localization to the Centrioles during Centriole Duplication*

PubMed Central

Gudi, Radhika; Zou, Chaozhong; Dhar, Jayeeta; Gao, Qingshen; Vasu, Chenthamarakshan

2014-01-01

Centriole duplication is the process by which two new daughter centrioles are generated from the proximal end of preexisting mother centrioles. Accurate centriole duplication is important for many cellular and physiological events, including cell division and ciliogenesis. Centrosomal protein 4.1-associated protein (CPAP), centrosomal protein of 152 kDa (CEP152), and centrobin are known to be essential for centriole duplication. However, the precise mechanism by which they contribute to centriole duplication is not known. In this study, we show that centrobin interacts with CEP152 and CPAP, and the centrobin-CPAP interaction is critical for centriole duplication. Although depletion of centrobin from cells did not have an effect on the centriolar levels of CEP152, it caused the disappearance of CPAP from both the preexisting and newly formed centrioles. Moreover, exogenous expression of the CPAP-binding fragment of centrobin also caused the disappearance of CPAP from both the preexisting and newly synthesized centrioles, possibly in a dominant negative manner, thereby inhibiting centriole duplication and the PLK4 overexpression-mediated centrosome amplification. Interestingly, exogenous overexpression of CPAP in the centrobin-depleted cells did not restore CPAP localization to the centrioles. However, restoration of centrobin expression in the centrobin-depleted cells led to the reappearance of centriolar CPAP. Hence, we conclude that centrobin-CPAP interaction is critical for the recruitment of CPAP to procentrioles to promote the elongation of daughter centrioles and for the persistence of CPAP on preexisting mother centrioles. Our study indicates that regulation of CPAP levels on the centrioles by centrobin is critical for preserving the normal size, shape, and number of centrioles in the cell. PMID:24700465

Cdc6 localizes to S- and G2-phase centrosomes in a cell cycle-dependent manner

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Gwang Su; Kang, Jeeheon; Bang, Sung Woong

2015-01-16

Highlights: • Cdc6 protein is a component of the pre-replicative complex required for chromosomal replication initiation. • Cdc6 localized to centrosomes of S and G2 phases in a cell cycle-dependent manner. • The centrosomal localization was governed by centrosomal localization signal sequences of Cdc6. • Deletions or substitution mutations on the centrosomal localization signal interfered with centrosomal localization of the Cdc6 proteins. - Abstract: The Cdc6 protein has been primarily investigated as a component of the pre-replicative complex for the initiation of chromosome replication, which contributes to maintenance of chromosomal integrity. Here, we show that Cdc6 localized to the centrosomesmore » during S and G2 phases of the cell cycle. The centrosomal localization was mediated by Cdc6 amino acid residues 311–366, which are conserved within other Cdc6 homologues and contains a putative nuclear export signal. Deletions or substitutions of the amino acid residues did not allow the proteins to localize to centrosomes. In contrast, DsRed tag fused to the amino acid residues localized to centrosomes. These results indicated that a centrosome localization signal is contained within amino acid residues 311–366. The cell cycle-dependent centrosomal localization of Cdc6 in S and G2 phases suggest a novel function of Cdc6 in centrosomes.« less

Epidermal development, growth control, and homeostasis in the face of centrosome amplification.

PubMed

Kulukian, Anita; Holland, Andrew J; Vitre, Benjamin; Naik, Shruti; Cleveland, Don W; Fuchs, Elaine

2015-11-17

As nucleators of the mitotic spindle and primary cilium, centrosomes play crucial roles in equal segregation of DNA content to daughter cells, coordination of growth and differentiation, and transduction of homeostatic cues. Whereas the majority of mammalian cells carry no more than two centrosomes per cell, exceptions to this rule apply in certain specialized tissues and in select disease states, including cancer. Centrosome amplification, or the condition of having more than two centrosomes per cell, has been suggested to contribute to instability of chromosomes, imbalance in asymmetric divisions, and reorganization of tissue architecture; however, the degree to which these conditions are a direct cause of or simply a consequence of human disease is poorly understood. Here we addressed this issue by generating a mouse model inducing centrosome amplification in a naturally proliferative epithelial tissue by elevating Polo-like kinase 4 (Plk4) expression in the skin epidermis. By altering centrosome numbers, we observed multiciliated cells, spindle orientation errors, and chromosome segregation defects within developing epidermis. None of these defects was sufficient to impart a proliferative advantage within the tissue, however. Rather, impaired mitoses led to p53-mediated cell death and contributed to defective growth and stratification. Despite these abnormalities, mice remained viable and healthy, although epidermal cells with centrosome amplification were still appreciable. Moreover, these abnormalities were insufficient to disrupt homeostasis and initiate or enhance tumorigenesis, underscoring the powerful surveillance mechanisms in the skin.

Aurora B Interaction of Centrosomal Nlp Regulates Cytokinesis*

PubMed Central

Yan, Jie; Jin, Shunqian; Li, Jia; Zhan, Qimin

2010-01-01

Cytokinesis is a fundamental cellular process, which ensures equal abscission and fosters diploid progenies. Aberrant cytokinesis may result in genomic instability and cell transformation. However, the underlying regulatory machinery of cytokinesis is largely undefined. Here, we demonstrate that Nlp (Ninein-like protein), a recently identified BRCA1-associated centrosomal protein that is required for centrosomes maturation at interphase and spindle formation in mitosis, also contributes to the accomplishment of cytokinesis. Through immunofluorescent analysis, Nlp is found to localize at midbody during cytokinesis. Depletion of endogenous Nlp triggers aborted division and subsequently leads to multinucleated phenotypes. Nlp can be recruited by Aurora B to the midbody apparatus via their physical association at the late stage of mitosis. Disruption of their interaction induces aborted cytokinesis. Importantly, Nlp is characterized as a novel substrate of Aurora B and can be phosphorylated by Aurora B. The specific phosphorylation sites are mapped at Ser-185, Ser-448, and Ser-585. The phosphorylation at Ser-448 and Ser-585 is likely required for Nlp association with Aurora B and localization at midbody. Meanwhile, the phosphorylation at Ser-185 is vital to Nlp protein stability. Disruptions of these phosphorylation sites abolish cytokinesis and lead to chromosomal instability. Collectively, these observations demonstrate that regulation of Nlp by Aurora B is critical for the completion of cytokinesis, providing novel insights into understanding the machinery of cell cycle progression. PMID:20864540

Aurora B interaction of centrosomal Nlp regulates cytokinesis.

PubMed

Yan, Jie; Jin, Shunqian; Li, Jia; Zhan, Qimin

2010-12-17

Cytokinesis is a fundamental cellular process, which ensures equal abscission and fosters diploid progenies. Aberrant cytokinesis may result in genomic instability and cell transformation. However, the underlying regulatory machinery of cytokinesis is largely undefined. Here, we demonstrate that Nlp (Ninein-like protein), a recently identified BRCA1-associated centrosomal protein that is required for centrosomes maturation at interphase and spindle formation in mitosis, also contributes to the accomplishment of cytokinesis. Through immunofluorescent analysis, Nlp is found to localize at midbody during cytokinesis. Depletion of endogenous Nlp triggers aborted division and subsequently leads to multinucleated phenotypes. Nlp can be recruited by Aurora B to the midbody apparatus via their physical association at the late stage of mitosis. Disruption of their interaction induces aborted cytokinesis. Importantly, Nlp is characterized as a novel substrate of Aurora B and can be phosphorylated by Aurora B. The specific phosphorylation sites are mapped at Ser-185, Ser-448, and Ser-585. The phosphorylation at Ser-448 and Ser-585 is likely required for Nlp association with Aurora B and localization at midbody. Meanwhile, the phosphorylation at Ser-185 is vital to Nlp protein stability. Disruptions of these phosphorylation sites abolish cytokinesis and lead to chromosomal instability. Collectively, these observations demonstrate that regulation of Nlp by Aurora B is critical for the completion of cytokinesis, providing novel insights into understanding the machinery of cell cycle progression.

Centrioles regulate centrosome size by controlling the rate of Cnn incorporation into the PCM.

PubMed

Conduit, Paul T; Brunk, Kathrin; Dobbelaere, Jeroen; Dix, Carly I; Lucas, Eliana P; Raff, Jordan W

2010-12-21

centrosomes are major microtubule organizing centers in animal cells, and they comprise a pair of centrioles surrounded by an amorphous pericentriolar material (PCM). Centrosome size is tightly regulated during the cell cycle, and it has recently been shown that the two centrosomes in certain stem cells are often asymmetric in size. There is compelling evidence that centrioles influence centrosome size, but how centrosome size is set remains mysterious. we show that the conserved Drosophila PCM protein Cnn exhibits an unusual dynamic behavior, because Cnn molecules only incorporate into the PCM closest to the centrioles and then spread outward through the rest of the PCM. Cnn incorporation into the PCM is driven by an interaction with the conserved centriolar proteins Asl (Cep152 in humans) and DSpd-2 (Cep192 in humans). The rate of Cnn incorporation into the PCM is tightly regulated during the cell cycle, and this rate influences the amount of Cnn in the PCM, which in turn is an important determinant of overall centrosome size. Intriguingly, daughter centrioles in syncytial embryos only start to incorporate Cnn as they disengage from their mothers; this generates a centrosome size asymmetry, with mother centrioles always initially organizing more Cnn than their daughters. centrioles can control the amount of PCM they organize by regulating the rate of Cnn incorporation into the PCM. This mechanism can explain how centrosome size is regulated during the cell cycle and also allows mother and daughter centrioles to set centrosome size independently of one another.

p53 Dependent Centrosome Clustering Prevents Multipolar Mitosis in Tetraploid Cells

PubMed Central

Yi, Qiyi; Zhao, Xiaoyu; Huang, Yun; Ma, Tieliang; Zhang, Yingyin; Hou, Heli; Cooke, Howard J.; Yang, Da-Qing; Wu, Mian; Shi, Qinghua

2011-01-01

Background p53 abnormality and aneuploidy often coexist in human tumors, and tetraploidy is considered as an intermediate between normal diploidy and aneuploidy. The purpose of this study was to investigate whether and how p53 influences the transformation from tetraploidy to aneuploidy. Principal Findings Live cell imaging was performed to determine the fates and mitotic behaviors of several human and mouse tetraploid cells with different p53 status, and centrosome and spindle immunostaining was used to investigate centrosome behaviors. We found that p53 dominant-negative mutation, point mutation, or knockout led to a 2∼ 33-fold increase of multipolar mitosis in N/TERT1, 3T3 and mouse embryonic fibroblasts (MEFs), while mitotic entry and cell death were not significantly affected. In p53-/- tetraploid MEFs, the ability of centrosome clustering was compromised, while centrosome inactivation was not affected. Suppression of RhoA/ROCK activity by specific inhibitors in p53-/- tetraploid MEFs enhanced centrosome clustering, decreased multipolar mitosis from 38% to 20% and 16% for RhoA and ROCK, respectively, while expression of constitutively active RhoA in p53+/+ tetraploid 3T3 cells increased the frequency of multipolar mitosis from 15% to 35%. Conclusions p53 could not prevent tetraploid cells entering mitosis or induce tetraploid cell death. However, p53 abnormality impaired centrosome clustering and lead to multipolar mitosis in tetraploid cells by modulating the RhoA/ROCK signaling pathway. PMID:22076149

Centrobin-centrosomal protein 4.1-associated protein (CPAP) interaction promotes CPAP localization to the centrioles during centriole duplication.

PubMed

Gudi, Radhika; Zou, Chaozhong; Dhar, Jayeeta; Gao, Qingshen; Vasu, Chenthamarakshan

2014-05-30

Centriole duplication is the process by which two new daughter centrioles are generated from the proximal end of preexisting mother centrioles. Accurate centriole duplication is important for many cellular and physiological events, including cell division and ciliogenesis. Centrosomal protein 4.1-associated protein (CPAP), centrosomal protein of 152 kDa (CEP152), and centrobin are known to be essential for centriole duplication. However, the precise mechanism by which they contribute to centriole duplication is not known. In this study, we show that centrobin interacts with CEP152 and CPAP, and the centrobin-CPAP interaction is critical for centriole duplication. Although depletion of centrobin from cells did not have an effect on the centriolar levels of CEP152, it caused the disappearance of CPAP from both the preexisting and newly formed centrioles. Moreover, exogenous expression of the CPAP-binding fragment of centrobin also caused the disappearance of CPAP from both the preexisting and newly synthesized centrioles, possibly in a dominant negative manner, thereby inhibiting centriole duplication and the PLK4 overexpression-mediated centrosome amplification. Interestingly, exogenous overexpression of CPAP in the centrobin-depleted cells did not restore CPAP localization to the centrioles. However, restoration of centrobin expression in the centrobin-depleted cells led to the reappearance of centriolar CPAP. Hence, we conclude that centrobin-CPAP interaction is critical for the recruitment of CPAP to procentrioles to promote the elongation of daughter centrioles and for the persistence of CPAP on preexisting mother centrioles. Our study indicates that regulation of CPAP levels on the centrioles by centrobin is critical for preserving the normal size, shape, and number of centrioles in the cell. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Distinct Sequence Elements of Cyclin B1 Promote Localization to Chromatin, Centrosomes, and Kinetochores during Mitosis

PubMed Central

Bentley, Anna M.; Normand, Guillaume; Hoyt, Jonathan

2007-01-01

The mitotic cyclins promote cell division by binding and activating cyclin-dependent kinases (CDKs). Each cyclin has a unique pattern of subcellular localization that plays a vital role in regulating cell division. During mitosis, cyclin B1 is known to localize to centrosomes, microtubules, and chromatin. To determine the mechanisms of cyclin B1 localization in M phase, we imaged full-length and mutant versions of human cyclin B1-enhanced green fluorescent protein in live cells by using spinning disk confocal microscopy. In addition to centrosome, microtubule, and chromatin localization, we found that cyclin B1 also localizes to unattached kinetochores after nuclear envelope breakdown. Kinetochore recruitment of cyclin B1 required the kinetochore proteins Hec1 and Mad2, and it was stimulated by microtubule destabilization. Mutagenesis studies revealed that cyclin B1 is recruited to kinetochores through both CDK1-dependent and -independent mechanisms. In contrast, localization of cyclin B1 to chromatin and centrosomes is independent of CDK1 binding. The N-terminal domain of cyclin B1 is necessary and sufficient for chromatin association, whereas centrosome recruitment relies on sequences within the cyclin box. Our data support a role for cyclin B1 function at unattached kinetochores, and they demonstrate that separable and distinct sequence elements target cyclin B1 to kinetochores, chromatin, and centrosomes during mitosis. PMID:17881737

The polarity protein Baz forms a platform for the centrosome orientation during asymmetric stem cell division in the Drosophila male germline.

PubMed

Inaba, Mayu; Venkei, Zsolt G; Yamashita, Yukiko M

2015-03-20

Many stem cells divide asymmetrically in order to balance self-renewal with differentiation. The essence of asymmetric cell division (ACD) is the polarization of cells and subsequent division, leading to unequal compartmentalization of cellular/extracellular components that confer distinct cell fates to daughter cells. Because precocious cell division before establishing cell polarity would lead to failure in ACD, these two processes must be tightly coupled; however, the underlying mechanism is poorly understood. In Drosophila male germline stem cells, ACD is prepared by stereotypical centrosome positioning. The centrosome orientation checkpoint (COC) further serves to ensure ACD by preventing mitosis upon centrosome misorientation. In this study, we show that Bazooka (Baz) provides a platform for the correct centrosome orientation and that Baz-centrosome association is the key event that is monitored by the COC. Our work provides a foundation for understanding how the correct cell polarity may be recognized by the cell to ensure productive ACD.

FancJ regulates interstrand crosslinker induced centrosome amplification through the activation of polo-like kinase 1

PubMed Central

Zou, Jianqiu; Tian, Fen; Li, Ji; Pickner, Wyatt; Long, Molly; Rezvani, Khosrow; Wang, Hongmin; Zhang, Dong

2013-01-01

Summary DNA damage response (DDR) and the centrosome cycle are two of the most critical processes for maintaining a stable genome in animals. Sporadic evidence suggests a connection between these two processes. Here, we report our findings that six Fanconi Anemia (FA) proteins, including FancI and FancJ, localize to the centrosome. Intriguingly, we found that the localization of FancJ to the mother centrosome is stimulated by a DNA interstrand crosslinker, Mitomycin C (MMC). We further show that, in addition to its role in interstrand crosslinking (ICL) repair, FancJ also regulates the normal centrosome cycle as well as ICL induced centrosome amplification by activating the polo-like kinase 1 (PLK1). We have uncovered a novel function of FancJ in centrosome biogenesis and established centrosome amplification as an integral part of the ICL response. PMID:24167712

Protein kinase C zeta suppresses low‐ or high‐grade colorectal cancer (CRC) phenotypes by interphase centrosome anchoring

PubMed Central

Deevi, Ravi Kiran; Javadi, Arman; McClements, Jane; Vohhodina, Jekaterina; Savage, Kienan; Loughrey, Maurice Bernard; Evergren, Emma

2018-01-01

Abstract Histological grading provides prognostic stratification of colorectal cancer (CRC) by scoring heterogeneous phenotypes. Features of aggressiveness include aberrant mitotic spindle configurations, chromosomal breakage, and bizarre multicellular morphology, but pathobiology is poorly understood. Protein kinase C zeta (PKCz) controls mitotic spindle dynamics, chromosome segregation, and multicellular patterns, but its role in CRC phenotype evolution remains unclear. Here, we show that PKCz couples genome segregation to multicellular morphology through control of interphase centrosome anchoring. PKCz regulates interdependent processes that control centrosome positioning. Among these, interaction between the cytoskeletal linker protein ezrin and its binding partner NHERF1 promotes the formation of a localized cue for anchoring interphase centrosomes to the cell cortex. Perturbation of these phenomena induced different outcomes in cells with single or extra centrosomes. Defective anchoring of a single centrosome promoted bipolar spindle misorientation, multi‐lumen formation, and aberrant epithelial stratification. Collectively, these disturbances induce cribriform multicellular morphology that is typical of some categories of low‐grade CRC. By contrast, defective anchoring of extra centrosomes promoted multipolar spindle formation, chromosomal instability (CIN), disruption of glandular morphology, and cell outgrowth across the extracellular matrix interface characteristic of aggressive, high‐grade CRC. Because PKCz enhances apical NHERF1 intensity in 3D epithelial cultures, we used an immunohistochemical (IHC) assay of apical NHERF1 intensity as an indirect readout of PKCz activity in translational studies. We show that apical NHERF1 IHC intensity is inversely associated with multipolar spindle frequency and high‐grade morphology in formalin‐fixed human CRC samples. To conclude, defective PKCz control of interphase centrosome anchoring may underlie

CENTROSOMES AND MICROTUBULES DURING MEIOSIS IN THE MUSHROOM BOLETUS RUBINELLUS

PubMed Central

McLaughlin, David J.

1971-01-01

The double centrosome in the basidium of Boletus rubinellus has been observed in three planes with the electron microscope at interphase preceding nuclear fusion, at prophase I, and at interphase I. It is composed of two components connected by a band-shaped middle part. At anaphase I a single, enlarged centrosome is found at the spindle pole, which is attached to the cell membrane. Microtubules mainly oriented parallel to the longitudinal axis of the basidium are present at prefusion, prophase I and interphase I. Cytoplasmic microtubules are absent when the spindle is present. The relationship of the centrosome in B. rubinellus to that in other organisms and the role of the cytoplasmic microtubules are discussed. PMID:4329156

Meier-Gorlin syndrome mutations disrupt an Orc1 CDK inhibitory domain and cause centrosome reduplication.

PubMed

Hossain, Manzar; Stillman, Bruce

2012-08-15

Like DNA replication, centrosomes are licensed to duplicate once per cell division cycle to ensure genetic stability. In addition to regulating DNA replication, the Orc1 subunit of the human origin recognition complex controls centriole and centrosome copy number. Here we report that Orc1 harbors a PACT centrosome-targeting domain and a separate domain that differentially inhibits the protein kinase activities of Cyclin E-CDK2 and Cyclin A-CDK2. A cyclin-binding motif (Cy motif) is required for Orc1 to bind Cyclin A and inhibit Cyclin A-CDK2 kinase activity but has no effect on Cyclin E-CDK2 kinase activity. In contrast, Orc1 inhibition of Cyclin E-CDK2 kinase activity occurs by a different mechanism that is affected by Orc1 mutations identified in Meier-Gorlin syndrome patients. The cyclin/CDK2 kinase inhibitory domain of Orc1, when tethered to the PACT domain, localizes to centrosomes and blocks centrosome reduplication. Meier-Gorlin syndrome mutations that disrupt Cyclin E-CDK2 kinase inhibition also allow centrosome reduplication. Thus, Orc1 contains distinct domains that control centrosome copy number and DNA replication. We suggest that the Orc1 mutations present in some Meier-Gorlin syndrome patients contribute to the pronounced microcephaly and dwarfism observed in these individuals by altering centrosome duplication in addition to DNA replication defects.

CDK5RAP2 Is an Essential Scaffolding Protein of the Corona of the Dictyostelium Centrosome.

PubMed

Pitzen, Valentin; Askarzada, Sophie; Gräf, Ralph; Meyer, Irene

2018-04-23

Dictyostelium centrosomes consist of a nucleus-associated cylindrical, three-layered core structure surrounded by a corona consisting of microtubule-nucleation complexes embedded in a scaffold of large coiled-coil proteins. One of them is the conserved CDK5RAP2 protein. Here we focus on the role of Dictyostelium CDK5RAP2 for maintenance of centrosome integrity, its interaction partners and its dynamic behavior during interphase and mitosis. GFP-CDK5RAP2 is present at the centrosome during the entire cell cycle except from a short period during prophase, correlating with the normal dissociation of the corona at this stage. RNAi depletion of CDK5RAP2 results in complete disorganization of centrosomes and microtubules suggesting that CDK5RAP2 is required for organization of the corona and its association to the core structure. This is in line with the observation that overexpressed GFP-CDK5RAP2 elicited supernumerary cytosolic MTOCs. The phenotype of CDK5RAP2 depletion was very reminiscent of that observed upon depletion of CP148, another scaffolding protein of the corona. BioID interaction assays revealed an interaction of CDK5RAP2 not only with the corona markers CP148, γ-tubulin, and CP248, but also with the core components Cep192, CP75, and CP91. Furthermore, protein localization studies in both depletion strains revealed that CP148 and CDK5RAP2 cooperate in corona organization.

Direct Microtubule-Binding by Myosin-10 Orients Centrosomes toward Retraction Fibers and Subcortical Actin Clouds.

PubMed

Kwon, Mijung; Bagonis, Maria; Danuser, Gaudenz; Pellman, David

2015-08-10

Positioning of centrosomes is vital for cell division and development. In metazoan cells, spindle positioning is controlled by a dynamic pool of subcortical actin that organizes in response to the position of retraction fibers. These actin "clouds" are proposed to generate pulling forces on centrosomes and mediate spindle orientation. However, the motors that pull astral microtubules toward these actin structures are not known. Here, we report that the unconventional myosin, Myo10, couples actin-dependent forces from retraction fibers and subcortical actin clouds to centrosomes. Myo10-mediated centrosome positioning requires its direct microtubule binding. Computational image analysis of large microtubule populations reveals a direct effect of Myo10 on microtubule dynamics and microtubule-cortex interactions. Myo10's role in centrosome positioning is distinct from, but overlaps with, that of dynein. Thus, Myo10 plays a key role in integrating the actin and microtubule cytoskeletons to position centrosomes and mitotic spindles. Copyright © 2015 Elsevier Inc. All rights reserved.

Direct Microtubule-Binding by Myosin-10 Orients Centrosomes toward Retraction Fibers and Subcortical Actin Clouds

PubMed Central

Kwon, Mijung; Bagonis, Maria; Danuser, Gaudenz; Pellman, David

2015-01-01

SUMMARY Positioning of centrosomes is vital for cell division and development. In metazoan cells, spindle positioning is controlled by a dynamic pool of subcortical actin that organizes in response to the position of retraction fibers. These actin “clouds” are proposed to generate pulling forces on centrosomes and mediate spindle orientation. However, the motors that pull astral microtubules toward these actin structures are not known. Here, we report that the unconventional myosin, Myo10, couples actin-dependent forces from retraction fibers and subcortical actin clouds to centrosomes. Myo10-mediated centrosome positioning requires its direct microtubule binding. Computational image analysis of large microtubule populations reveals a direct effect of Myo10 on microtubule dynamics and microtubule-cortex interactions. Myo10’s role in centrosome positioning is distinct from, but overlaps with, that of dynein. Thus, Myo10 plays a key role in integrating the actin and microtubule cytoskeletons to position centrosomes and mitotic spindles. PMID:26235048

Structural centrosome aberrations favor proliferation by abrogating microtubule-dependent tissue integrity of breast epithelial mammospheres

PubMed Central

Schnerch, D; Nigg, E A

2016-01-01

Structural centrosome aberrations are frequently observed in early stage carcinomas, but their role in malignant transformation is poorly understood. Here, we examined the impact of overexpression of Ninein-like protein (Nlp) on the architecture of polarized epithelia in three-dimensional mammospheres. When Nlp was overexpressed to levels resembling those seen in human tumors, it formed striking centrosome-related bodies (CRBs), which sequestered Ninein and affected the kinetics of microtubule (MT) nucleation and release. In turn, the profound reorganization of the MT cytoskeleton resulted in mislocalization of several adhesion and junction proteins as well as the tumor suppressor Scribble, resulting in the disruption of epithelial polarity, cell-cell interactions and mammosphere architecture. Remarkably, cells harboring Nlp-CRBs displayed an enhanced proliferative response to epidermal growth factor. These results demonstrate that structural centrosome aberrations cause not only the disruption of epithelial polarity but also favor overproliferation, two phenotypes typically associated with human carcinomas. PMID:26364601

Conserved TCP domain of Sas-4/CPAP is essential for pericentriolar material tethering during centrosome biogenesis

PubMed Central

Zheng, Xiangdong; Gooi, Li Ming; Wason, Arpit; Gabriel, Elke; Mehrjardi, Narges Zare; Yang, Qian; Zhang, Xingrun; Debec, Alain; Basiri, Marcus L.; Avidor-Reiss, Tomer; Pozniakovsky, Andrei; Poser, Ina; Šarić, Tomo; Hyman, Anthony A.; Li, Haitao; Gopalakrishnan, Jay

2014-01-01

Pericentriolar material (PCM) recruitment to centrioles forms a key step in centrosome biogenesis. Deregulation of this process leads to centrosome aberrations causing disorders, one of which is autosomal recessive primary microcephaly (MCPH), a neurodevelopmental disorder where brain size is reduced. During PCM recruitment, the conserved centrosomal protein Sas-4/CPAP/MCPH6, known to play a role in centriole formation, acts as a scaffold for cytoplasmic PCM complexes to bind and then tethers them to centrioles to form functional centrosomes. To understand Sas-4’s tethering role, we determined the crystal structure of its T complex protein 10 (TCP) domain displaying a solvent-exposed single-layer of β-sheets fold. This unique feature of the TCP domain suggests that it could provide an “extended surface-like” platform to tether the Sas-4–PCM scaffold to a centriole. Functional studies in Drosophila, human cells, and human induced pluripotent stem cell-derived neural progenitor cells were used to test this hypothesis, where point mutations within the 9–10th β-strands (β9–10 mutants including a MCPH-associated mutation) perturbed PCM tethering while allowing Sas-4/CPAP to scaffold cytoplasmic PCM complexes. Specifically, the Sas-4 β9–10 mutants displayed perturbed interactions with Ana2, a centrosome duplication factor, and Bld-10, a centriole microtubule-binding protein, suggesting a role for the β9–10 surface in mediating protein–protein interactions for efficient Sas-4–PCM scaffold centriole tethering. Hence, we provide possible insights into how centrosomal protein defects result in human MCPH and how Sas-4 proteins act as a vehicle to tether PCM complexes to centrioles independent of its well-known role in centriole duplication. PMID:24385583

Centrosome misorientation mediates slowing of the cell cycle under limited nutrient conditions in Drosophila male germline stem cells

PubMed Central

Roth, Therese M.; Chiang, C.-Y. Ason; Inaba, Mayu; Yuan, Hebao; Salzmann, Viktoria; Roth, Caitlin E.; Yamashita, Yukiko M.

2012-01-01

Drosophila male germline stem cells (GSCs) divide asymmetrically, balancing self-renewal and differentiation. Although asymmetric stem cell division balances between self-renewal and differentiation, it does not dictate how frequently differentiating cells must be produced. In male GSCs, asymmetric GSC division is achieved by stereotyped positioning of the centrosome with respect to the stem cell niche. Recently we showed that the centrosome orientation checkpoint monitors the correct centrosome orientation to ensure an asymmetric outcome of the GSC division. When GSC centrosomes are not correctly oriented with respect to the niche, GSC cell cycle is arrested/delayed until the correct centrosome orientation is reacquired. Here we show that induction of centrosome misorientation upon culture in poor nutrient conditions mediates slowing of GSC cell proliferation via activation of the centrosome orientation checkpoint. Consistently, inactivation of the centrosome orientation checkpoint leads to lack of cell cycle slowdown even under poor nutrient conditions. We propose that centrosome misorientation serves as a mediator that transduces nutrient information into stem cell proliferation, providing a previously unappreciated mechanism of stem cell regulation in response to nutrient conditions. PMID:22357619

Drosophila parthenogenesis: A tool to decipher centrosomal vs acentrosomal spindle assembly pathways

DOE Office of Scientific and Technical Information (OSTI.GOV)

Riparbelli, Maria Giovanna; Callaini, Giuliano

2008-04-15

Development of unfertilized eggs in the parthenogenetic strain K23-O-im of Drosophila mercatorum requires the stochastic interactions of self-assembled centrosomes with the female chromatin. In a portion of the unfertilized eggs that do not assemble centrosomes, microtubules organize a bipolar anastral mitotic spindle around the chromatin like the one formed during the first female meiosis, suggesting that similar pathways may be operative. In the cytoplasm of eggs in which centrosomes do form, monastral and biastral spindles are found. Analysis by laser scanning confocal microscopy suggests that these spindles are derived from the stochastic interaction of astral microtubules directly with kinetochore regionsmore » or indirectly with kinetochore microtubules. Our findings are consistent with the idea that mitotic spindle assembly requires both acentrosomal and centrosomal pathways, strengthening the hypothesis that astral microtubules can dictate the organization of the spindle by capturing kinetochore microtubules.« less

Protein kinase C zeta suppresses low- or high-grade colorectal cancer (CRC) phenotypes by interphase centrosome anchoring.

PubMed

Deevi, Ravi Kiran; Javadi, Arman; McClements, Jane; Vohhodina, Jekaterina; Savage, Kienan; Loughrey, Maurice Bernard; Evergren, Emma; Campbell, Frederick Charles

2018-04-01

Histological grading provides prognostic stratification of colorectal cancer (CRC) by scoring heterogeneous phenotypes. Features of aggressiveness include aberrant mitotic spindle configurations, chromosomal breakage, and bizarre multicellular morphology, but pathobiology is poorly understood. Protein kinase C zeta (PKCz) controls mitotic spindle dynamics, chromosome segregation, and multicellular patterns, but its role in CRC phenotype evolution remains unclear. Here, we show that PKCz couples genome segregation to multicellular morphology through control of interphase centrosome anchoring. PKCz regulates interdependent processes that control centrosome positioning. Among these, interaction between the cytoskeletal linker protein ezrin and its binding partner NHERF1 promotes the formation of a localized cue for anchoring interphase centrosomes to the cell cortex. Perturbation of these phenomena induced different outcomes in cells with single or extra centrosomes. Defective anchoring of a single centrosome promoted bipolar spindle misorientation, multi-lumen formation, and aberrant epithelial stratification. Collectively, these disturbances induce cribriform multicellular morphology that is typical of some categories of low-grade CRC. By contrast, defective anchoring of extra centrosomes promoted multipolar spindle formation, chromosomal instability (CIN), disruption of glandular morphology, and cell outgrowth across the extracellular matrix interface characteristic of aggressive, high-grade CRC. Because PKCz enhances apical NHERF1 intensity in 3D epithelial cultures, we used an immunohistochemical (IHC) assay of apical NHERF1 intensity as an indirect readout of PKCz activity in translational studies. We show that apical NHERF1 IHC intensity is inversely associated with multipolar spindle frequency and high-grade morphology in formalin-fixed human CRC samples. To conclude, defective PKCz control of interphase centrosome anchoring may underlie distinct categories of

Amphiastral Mitotic Spindle Assembly in Vertebrate Cells Lacking Centrosomes

PubMed Central

Hornick, Jessica E.; Mader, Christopher C.; Tribble, Emily K.; Bagne, Cydney C.; Vaughan, Kevin T.; Shaw, Sidney L.; Hinchcliffe, Edward H.

2011-01-01

Summary The role of centrosomes/centrioles during mitotic spindle assembly in vertebrates remains controversial. In cell-free extracts and experimentally derived acentrosomal cells, randomly oriented microtubules (MTs) self-organize around mitotic chromosomes and assemble anastral spindles [1, 2, 3]. However, vertebrate somatic cells normally assemble a connected pair of polarized, astral MT arrays – termed an amphiaster (“a star on both sides” [4]) – that is formed by the splitting and separation of the microtubule-organizing center (MTOC) well before nuclear envelope breakdown (NEB) [5]. Whether amphiaster formation requires splitting of duplicated centrosomes is not known. We found that when centrosomes were removed from living vertebrate cells early in their cell cycle, an acentriolar MTOC re-assembled, and prior to NEB, a functional amphiastral spindle formed. Cytoplasmic dynein, dynactin, and pericentrin are all recruited to the interphase aMTOC, and the activity of kinesin-5 is needed for amphiaster formation. Mitosis proceeded on time and these karyoplasts divided in two. However, ~35% of aMTOCs failed to split/separate before NEB, and these entered mitosis with persistent monastral spindles. The chromatin-mediated RAN-GTP pathway could not restore bipolarity to monastral spindles, and these cells exited mitosis as single daughters. Our data reveal the novel finding that MTOC separation and amphiaster formation does not absolutely require the centrosome, but in its absence, the fidelity of bipolar spindle assembly is highly compromised. PMID:21439826

Cleavage of the SUN-domain protein Mps3 at its N-terminus regulates centrosome disjunction in budding yeast meiosis

PubMed Central

Koch, Bailey A.; Han, Xuemei

2017-01-01

Centrosomes organize microtubules and are essential for spindle formation and chromosome segregation during cell division. Duplicated centrosomes are physically linked, but how this linkage is dissolved remains unclear. Yeast centrosomes are tethered by a nuclear-envelope-attached structure called the half-bridge, whose components have mammalian homologues. We report here that cleavage of the half-bridge protein Mps3 promotes accurate centrosome disjunction in budding yeast. Mps3 is a single-pass SUN-domain protein anchored at the inner nuclear membrane and concentrated at the nuclear side of the half-bridge. Using the unique feature in yeast meiosis that centrosomes are linked for hours before their separation, we have revealed that Mps3 is cleaved at its nucleus-localized N-terminal domain, the process of which is regulated by its phosphorylation at serine 70. Cleavage of Mps3 takes place at the yeast centrosome and requires proteasome activity. We show that noncleavable Mps3 (Mps3-nc) inhibits centrosome separation during yeast meiosis. In addition, overexpression of mps3-nc in vegetative yeast cells also inhibits centrosome separation and is lethal. Our findings provide a genetic mechanism for the regulation of SUN-domain protein-mediated activities, including centrosome separation, by irreversible protein cleavage at the nuclear periphery. PMID:28609436

Centrin 3 is an inhibitor of centrosomal Mps1 and antagonizes centrin 2 function

PubMed Central

Sawant, Dwitiya B.; Majumder, Shubhra; Perkins, Jennifer L.; Yang, Ching-Hui; Eyers, Patrick A.; Fisk, Harold A.

2015-01-01

Centrins are a family of small, calcium-binding proteins with diverse cellular functions that play an important role in centrosome biology. We previously identified centrin 2 and centrin 3 (Cetn2 and Cetn3) as substrates of the protein kinase Mps1. However, although Mps1 phosphorylation sites control the function of Cetn2 in centriole assembly and promote centriole overproduction, Cetn2 and Cetn3 are not functionally interchangeable, and we show here that Cetn3 is both a biochemical inhibitor of Mps1 catalytic activity and a biological inhibitor of centrosome duplication. In vitro, Cetn3 inhibits Mps1 autophosphorylation at Thr-676, a known site of T-loop autoactivation, and interferes with Mps1-dependent phosphorylation of Cetn2. The cellular overexpression of Cetn3 attenuates the incorporation of Cetn2 into centrioles and centrosome reduplication, whereas depletion of Cetn3 generates extra centrioles. Finally, overexpression of Cetn3 reduces Mps1 Thr-676 phosphorylation at centrosomes, and mimicking Mps1-dependent phosphorylation of Cetn2 bypasses the inhibitory effect of Cetn3, suggesting that the biological effects of Cetn3 are due to the inhibition of Mps1 function at centrosomes. PMID:26354417

Luna, a Drosophila KLF6/KLF7, Is Maternally Required for Synchronized Nuclear and Centrosome Cycles in the Preblastoderm Embryo

PubMed Central

Weber, Ursula; Rodriguez, Estefania; Martignetti, John; Mlodzik, Marek

2014-01-01

Krüppel like factors (KLFs) are conserved transcription factors that have been implicated in many developmental processes including differentiation, organ patterning, or regulation of stem cell pluripotency. We report the generation and analysis of loss-of-function mutants of Drosophila Klf6/7, the luna gene. We demonstrate that luna mutants are associated with very early embryonic defects prior to cellularization at the syncytial stage and cause DNA separation defects during the rapid mitotic cycles resulting in un-coupled DNA and centrosome cycles. These defects manifest themselves, both in animals that are maternally homozygous and heterozygous mutant. Surprisingly, luna is only required during the syncytial stages and not later in development, suggesting that the DNA segregation defect is linked to centrosomes, since centrosomes are dispensable for later cell divisions. PMID:24915236

The C. elegans RSA complex localizes protein phosphatase 2A to centrosomes and regulates mitotic spindle assembly.

PubMed

Schlaitz, Anne-Lore; Srayko, Martin; Dammermann, Alexander; Quintin, Sophie; Wielsch, Natalie; MacLeod, Ian; de Robillard, Quentin; Zinke, Andrea; Yates, John R; Müller-Reichert, Thomas; Shevchenko, Andrei; Oegema, Karen; Hyman, Anthony A

2007-01-12

Microtubule behavior changes during the cell cycle and during spindle assembly. However, it remains unclear how these changes are regulated and coordinated. We describe a complex that targets the Protein Phosphatase 2A holoenzyme (PP2A) to centrosomes in C. elegans embryos. This complex includes Regulator of Spindle Assembly 1 (RSA-1), a targeting subunit for PP2A, and RSA-2, a protein that binds and recruits RSA-1 to centrosomes. In contrast to the multiple functions of the PP2A catalytic subunit, RSA-1 and RSA-2 are specifically required for microtubule outgrowth from centrosomes and for spindle assembly. The centrosomally localized RSA-PP2A complex mediates these functions in part by regulating two critical mitotic effectors: the microtubule destabilizer KLP-7 and the C. elegans regulator of spindle assembly TPXL-1. By regulating a subset of PP2A functions at the centrosome, the RSA complex could therefore provide a means of coordinating microtubule outgrowth from centrosomes and kinetochore microtubule stability during mitotic spindle assembly.

Excess free histone H3 localizes to centrosomes for proteasome-mediated degradation during mitosis in metazoans.

PubMed

Wike, Candice L; Graves, Hillary K; Wason, Arpit; Hawkins, Reva; Gopalakrishnan, Jay; Schumacher, Jill; Tyler, Jessica K

2016-08-17

The cell tightly controls histone protein levels in order to achieve proper packaging of the genome into chromatin, while avoiding the deleterious consequences of excess free histones. Our accompanying study has shown that a histone modification that loosens the intrinsic structure of the nucleosome, phosphorylation of histone H3 on threonine 118 (H3 T118ph), exists on centromeres and chromosome arms during mitosis. Here, we show that H3 T118ph localizes to centrosomes in humans, flies, and worms during all stages of mitosis. H3 abundance at the centrosome increased upon proteasome inhibition, suggesting that excess free histone H3 localizes to centrosomes for degradation during mitosis. In agreement, we find ubiquitinated H3 specifically during mitosis and within purified centrosomes. These results suggest that targeting of histone H3 to the centrosome for proteasome-mediated degradation is a novel pathway for controlling histone supply, specifically during mitosis.

Centrosome structure and function is altered by chloral hydrate and diazepam during the first reproductive cell cycles in sea urchin eggs

NASA Technical Reports Server (NTRS)

Schatten, H.; Chakrabarti, A.

1998-01-01

This paper explores the mode of action of the tranquillizers chloral hydrate and diazepam during fertilization and mitosis of the first reproductive cell cycles in sea urchin eggs. Most striking effects of these drugs are the alteration of centrosomal material and the abnormal microtubule configurations during exposure and after recovery from the drugs. This finding is utilized to study the mechanisms of centrosome compaction and decompaction and the dynamic configurational changes of centrosomal material and its interactions with microtubules. When 0.1% chloral hydrate or 350-750 microM diazepam is applied at specific phases during the first cell cycle of sea urchin eggs, expanded centrosomal material compacts at distinct regions and super-compacts into dense spheres while microtubules disassemble. When eggs are treated before pronuclear fusion, centrosomal material aggregates around each of the two pronuclei while microtubules disappear. Upon recovery, atypical asters oftentimes with multiple foci are formed from centrosomal material surrounding the pronuclei which indicates that the drugs have affected centrosomal material and prevent it from functioning normally. Electron microscopy and immunofluorescence studies with antibodies that routinely stain centrosomes in sea urchin eggs (4D2; and Ah-6) depict centrosomal material that is altered when compared to control cells. This centrosomal material is not able to reform normal microtubule patterns upon recovery but will form multiple asters around the two pronuclei. When cells are treated with 0.1% chloral hydrate or 350-750 microM diazepam during mitosis, the bipolar centrosomal material becomes compacted and aggregates into multiple dense spheres while spindle and polar microtubules disassemble. With increased incubation time, the smaller dense centrosome particles aggregate into bigger and fewer spheres. Upon recovery, unusual irregular microtubule configurations are formed from centrosomes that have lost their

Transcriptional program of ciliated epithelial cells reveals new cilium and centrosome components and links to human disease.

PubMed

Hoh, Ramona A; Stowe, Timothy R; Turk, Erin; Stearns, Tim

2012-01-01

Defects in the centrosome and cilium are associated with a set of human diseases having diverse phenotypes. To further characterize the components that define the function of these organelles we determined the transcriptional profile of multiciliated tracheal epithelial cells. Cultures of mouse tracheal epithelial cells undergoing differentiation in vitro were derived from mice expressing GFP from the ciliated-cell specific FOXJ1 promoter (FOXJ1:GFP). The transcriptional profile of ciliating GFP+ cells from these cultures was defined at an early and a late time point during differentiation and was refined by subtraction of the profile of the non-ciliated GFP- cells. We identified 649 genes upregulated early, when most cells were forming basal bodies, and 73 genes genes upregulated late, when most cells were fully ciliated. Most, but not all, of known centrosome proteins are transcriptionally upregulated early, particularly Plk4, a master regulator of centriole formation. We found that three genes associated with human disease states, Mdm1, Mlf1, and Dyx1c1, are upregulated during ciliogenesis and localize to centrioles and cilia. This transcriptome for mammalian multiciliated epithelial cells identifies new candidate centrosome and cilia proteins, highlights similarities between components of motile and primary cilia, and identifies new links between cilia proteins and human disease.

Transcriptional Program of Ciliated Epithelial Cells Reveals New Cilium and Centrosome Components and Links to Human Disease

PubMed Central

Hoh, Ramona A.; Stowe, Timothy R.; Turk, Erin; Stearns, Tim

2012-01-01

Defects in the centrosome and cilium are associated with a set of human diseases having diverse phenotypes. To further characterize the components that define the function of these organelles we determined the transcriptional profile of multiciliated tracheal epithelial cells. Cultures of mouse tracheal epithelial cells undergoing differentiation in vitro were derived from mice expressing GFP from the ciliated-cell specific FOXJ1 promoter (FOXJ1:GFP). The transcriptional profile of ciliating GFP+ cells from these cultures was defined at an early and a late time point during differentiation and was refined by subtraction of the profile of the non-ciliated GFP- cells. We identified 649 genes upregulated early, when most cells were forming basal bodies, and 73 genes genes upregulated late, when most cells were fully ciliated. Most, but not all, of known centrosome proteins are transcriptionally upregulated early, particularly Plk4, a master regulator of centriole formation. We found that three genes associated with human disease states, Mdm1, Mlf1, and Dyx1c1, are upregulated during ciliogenesis and localize to centrioles and cilia. This transcriptome for mammalian multiciliated epithelial cells identifies new candidate centrosome and cilia proteins, highlights similarities between components of motile and primary cilia, and identifies new links between cilia proteins and human disease. PMID:23300604

Cnn dynamics drive centrosome size asymmetry to ensure daughter centriole retention in Drosophila neuroblasts.

PubMed

Conduit, Paul T; Raff, Jordan W

2010-12-21

Centrosomes comprise a pair of centrioles surrounded by an amorphous network of pericentriolar material (PCM). In certain stem cells, the two centrosomes differ in size, and this appears to be important for asymmetric cell division [1, 2]. In some cases, centrosome asymmetry is linked to centriole age because the older, mother centriole always organizes more PCM than the daughter centriole, thus ensuring that the mother centriole is always retained in the stem cell after cell division [3]. This has raised the possibility that an "immortal" mother centriole may help maintain stem cell fate [4, 5]. It is unclear, however, how centrosome size asymmetry is generated in stem cells. Here we provide compelling evidence that centrosome size asymmetry in Drosophila neuroblasts is generated by the differential regulation of Cnn incorporation into the PCM at mother and daughter centrioles. Shortly after centriole separation, mother and daughter centrioles organize similar amounts of PCM, but Cnn incorporation is then rapidly downregulated at the mother centriole, while it is maintained at the daughter centriole. This ensures that the daughter centriole maintains its PCM and so its position at the apical cortex. Thus, the daughter centriole, rather than an "immortal" mother centriole, is ultimately retained in these stem cells.

Inter-dependent apical microtubule and actin dynamics orchestrate centrosome retention and neuronal delamination

PubMed Central

Kasioulis, Ioannis

2017-01-01

Detachment of newborn neurons from the neuroepithelium is required for correct neuronal architecture and functional circuitry. This process, also known as delamination, involves adherens-junction disassembly and acto-myosin-mediated abscission, during which the centrosome is retained while apical/ciliary membranes are shed. Cell-biological mechanisms mediating delamination are, however, poorly understood. Using live-tissue and super-resolution imaging, we uncover a centrosome-nucleated wheel-like microtubule configuration, aligned with the apical actin cable and adherens-junctions within chick and mouse neuroepithelial cells. These microtubules maintain adherens-junctions while actin maintains microtubules, adherens-junctions and apical end-foot dimensions. During neuronal delamination, acto-myosin constriction generates a tunnel-like actin-microtubule configuration through which the centrosome translocates. This movement requires inter-dependent actin and microtubule activity, and we identify drebrin as a potential coordinator of these cytoskeletal dynamics. Furthermore, centrosome compromise revealed that this organelle is required for delamination. These findings identify new cytoskeletal configurations and regulatory relationships that orchestrate neuronal delamination and may inform mechanisms underlying pathological epithelial cell detachment. PMID:29058679

Perturbation of nucleo-cytoplasmic transport affects size of nucleus and nucleolus in human cells.

PubMed

Ganguly, Abira; Bhattacharjee, Chumki; Bhave, Madhura; Kailaje, Vaishali; Jain, Bhawik K; Sengupta, Isha; Rangarajan, Annapoorni; Bhattacharyya, Dibyendu

2016-03-01

Size regulation of human cell nucleus and nucleolus are poorly understood subjects. 3D reconstruction of live image shows that the karyoplasmic ratio (KR) increases by 30-80% in transformed cell lines compared to their immortalized counterpart. The attenuation of nucleo-cytoplasmic transport causes the KR value to increase by 30-50% in immortalized cell lines. Nucleolus volumes are significantly increased in transformed cell lines and the attenuation of nucleo-cytoplasmic transport causes a significant increase in the nucleolus volume of immortalized cell lines. A cytosol and nuclear fraction swapping experiment emphasizes the potential role of unknown cytosolic factors in nuclear and nucleolar size regulation. © 2016 Federation of European Biochemical Societies.

Recruitment of PCM1 to the centrosome by the cooperative action of DISC1 and BBS4: a candidate for psychiatric illnesses.

PubMed

Kamiya, Atsushi; Tan, Perciliz L; Kubo, Ken-ichiro; Engelhard, Caitlin; Ishizuka, Koko; Kubo, Akiharu; Tsukita, Sachiko; Pulver, Ann E; Nakajima, Kazunori; Cascella, Nicola G; Katsanis, Nicholas; Sawa, Akira

2008-09-01

A role for the centrosome has been suggested in the pathology of major mental illnesses, especially schizophrenia (SZ). To show that pericentriolar material 1 protein (PCM1) forms a complex at the centrosome with disrupted-in-schizophrenia 1 (DISC1) and Bardet-Biedl syndrome 4 protein (BBS4), which provides a crucial pathway for cortical development associated with the pathology of SZ. To identify mutations in the PCM1 gene in an SZ population. Interaction of DISC1, PCM1, and BBS proteins was assessed by immunofluorescent staining and coimmunoprecipitation. Effects of PCM1, DISC1, and BBS on centrosomal functions and corticogenesis in vivo were tested by RNA interference. The PCM1 gene was examined by sequencing 39 exons and flanking splice sites. Probands and controls were from the collection of one of us (A.E.P.). Thirty-two probands with SZ from families that had excess allele sharing among affected individuals at 8p22 and 219 white controls. Protein interaction and recruitment at the centrosome in cells; neuronal migration in the cerebral cortex; and variant discovery in PCM1 in patients with SZ. PCM1 forms a complex with DISC1 and BBS4 through discrete binding domains in each protein. DISC1 and BBS4 are required for targeting PCM1 and other cargo proteins, such as ninein, to the centrosome in a synergistic manner. In the developing cerebral cortex, suppression of PCM1 leads to neuronal migration defects, which are phenocopied by the suppression of either DISC1 or BBS4 and are exacerbated by the concomitant suppression of both. Furthermore, a nonsense mutation that segregates with SZ spectrum psychosis was found in 1 family. Our data further support for the role of centrosomal proteins in cortical development and suggest that perturbation of centrosomal function contributes to the development of mental diseases, including SZ.

Novel microcephalic primordial dwarfism disorder associated with variants in the centrosomal protein ninein.

PubMed

Dauber, Andrew; Lafranchi, Stephen H; Maliga, Zoltan; Lui, Julian C; Moon, Jennifer E; McDeed, Cailin; Henke, Katrin; Zonana, Jonathan; Kingman, Garrett A; Pers, Tune H; Baron, Jeffrey; Rosenfeld, Ron G; Hirschhorn, Joel N; Harris, Matthew P; Hwa, Vivian

2012-11-01

Microcephalic primordial dwarfism (MPD) is a rare, severe form of human growth failure in which growth restriction is evident in utero and continues into postnatal life. Single causative gene defects have been identified in a number of patients with MPD, and all involve genes fundamental to cellular processes including centrosome functions. The objective of the study was to find the genetic etiology of a novel presentation of MPD. The design of the study was whole-exome sequencing performed on two affected sisters in a single family. Molecular and functional studies of a candidate gene were performed using patient-derived primary fibroblasts and a zebrafish morpholino oligonucleotides knockdown model. Two sisters presented with a novel subtype of MPD, including severe intellectual disabilities. NIN, encoding Ninein, a centrosomal protein critically involved in asymmetric cell division, was identified as a candidate gene, and functional impacts in fibroblasts and zebrafish were studied. From 34,606 genomic variants, two very rare missense variants in NIN were identified. Both probands were compound heterozygotes. In the zebrafish, ninein knockdown led to specific and novel defects in the specification and morphogenesis of the anterior neuroectoderm, resulting in a deformity of the developing cranium with a small, squared skull highly reminiscent of the human phenotype. We identified a novel clinical subtype of MPD in two sisters who have rare variants in NIN. We show, for the first time, that reduction of ninein function in the developing zebrafish leads to specific deficiencies of brain and skull development, offering a developmental basis for the myriad phenotypes in our patients.

Cdc2/cyclin B1 regulates centrosomal Nlp proteolysis and subcellular localization.

PubMed

Zhao, Xuelian; Jin, Shunqian; Song, Yongmei; Zhan, Qimin

2010-11-01

The formation of proper mitotic spindles is required for appropriate chromosome segregation during cell division. Aberrant spindle formation often causes aneuploidy and results in tumorigenesis. However, the underlying mechanism of regulating spindle formation and chromosome separation remains to be further defined. Centrosomal Nlp (ninein-like protein) is a recently characterized BRCA1-regulated centrosomal protein and plays an important role in centrosome maturation and spindle formation. In this study, we show that Nlp can be phosphorylated by cell cycle protein kinase Cdc2/cyclin B1. The phosphorylation sites of Nlp are mapped at Ser185 and Ser589. Interestingly, the Cdc2/cyclin B1 phosphorylation site Ser185 of Nlp is required for its recognition by PLK1, which enable Nlp depart from centrosomes to allow the establishment of a mitotic scaffold at the onset of mitosis . PLK1 fails to dissociate the Nlp mutant lacking Ser185 from centrosome, suggesting that Cdc2/cyclin B1 might serve as a primary kinase of PLK1 in regulating Nlp subcellular localization. However, the phosphorylation at the site Ser589 by Cdc2/cyclin B1 plays an important role in Nlp protein stability probably due to its effect on protein degradation. Furthermore, we show that deregulated expression or subcellular localization of Nlp lead to multinuclei in cells, indicating that scheduled levels of Nlp and proper subcellular localization of Nlp are critical for successful completion of normal cell mitosis, These findings demonstrate that Cdc2/cyclin B1 is a key regulator in maintaining appropriate degradation and subcellular localization of Nlp, providing novel insights into understanding on the role of Cdc2/cyclin B1 in mitotic progression.

Cdk1 phosphorylates the Rac activator Tiam1 to activate centrosomal Pak and promote mitotic spindle formation

PubMed Central

Whalley, Helen J.; Porter, Andrew P.; Diamantopoulou, Zoi; White, Gavin R. M.; Castañeda-Saucedo, Eduardo; Malliri, Angeliki

2015-01-01

Centrosome separation is critical for bipolar spindle formation and the accurate segregation of chromosomes during mammalian cell mitosis. Kinesin-5 (Eg5) is a microtubule motor essential for centrosome separation, and Tiam1 and its substrate Rac antagonize Eg5-dependent centrosome separation in early mitosis promoting efficient chromosome congression. Here we identify S1466 of Tiam1 as a novel Cdk1 site whose phosphorylation is required for the mitotic function of Tiam1. We find that this phosphorylation of Tiam1 is required for the activation of group I p21-activated kinases (Paks) on centrosomes in prophase. Further, we show that both Pak1 and Pak2 counteract centrosome separation in a kinase-dependent manner and demonstrate that they act downstream of Tiam1. We also show that depletion of Pak1/2 allows cells to escape monopolar arrest by Eg5 inhibition, highlighting the potential importance of this signalling pathway for the development of Eg5 inhibitors as cancer therapeutics. PMID:26078008

Hierarchical assembly of centriole subdistal appendages via centrosome binding proteins CCDC120 and CCDC68.

PubMed

Huang, Ning; Xia, Yuqing; Zhang, Donghui; Wang, Song; Bao, Yitian; He, Runsheng; Teng, Junlin; Chen, Jianguo

2017-04-19

In animal cells, the centrosome is the main microtubule-organizing centre where microtubules are nucleated and anchored. The centriole subdistal appendages (SDAs) are the key structures that anchor microtubules in interphase cells, but the composition and assembly mechanisms of SDAs are not well understood. Here, we reveal that centrosome-binding proteins, coiled-coil domain containing (CCDC) 120 and CCDC68 are two novel SDA components required for hierarchical SDA assembly in human cells. CCDC120 is anchored to SDAs by ODF2 and recruits CEP170 and Ninein to the centrosome through different coiled-coil domains at its N terminus. CCDC68 is a CEP170-interacting protein that competes with CCDC120 in recruiting CEP170 to SDAs. Furthermore, CCDC120 and CCDC68 are required for centrosome microtubule anchoring. Our findings elucidate the molecular basis for centriole SDA hierarchical assembly and microtubule anchoring in human interphase cells.

Historical roots of centrosome research: discovery of Boveri's microscope slides in Würzburg

PubMed Central

Scheer, Ulrich

2014-01-01

Boveri's visionary monograph ‘Ueber die Natur der Centrosomen’ (On the nature of centrosomes) in 1900 was founded primarily on microscopic observations of cleaving eggs of sea urchins and the roundworm parasite Ascaris. As Boveri wrote in the introductory paragraph, his interests were less about morphological aspects of centrosomes, but rather aimed at an understanding of their physiological role during cell division. The remarkable transition from observations of tiny dot-like structures in fixed and sectioned material to a unified theory of centrosome function (which in essence still holds true today) cannot be fully appreciated without examining Boveri's starting material, the histological specimens. It was generally assumed that the microscope slides were lost during the bombing of the Zoological Institute in Würzburg at the end of WWII. Here, I describe the discovery of a number of Boveri's original microscope slides with serial sections of early sea urchin and Ascaris embryos, stained by Heidenhain's iron haematoxylin method. Some slides bear handwritten notes and sketches by Boveri. Evidence is presented that the newly discovered slides are part of the original material used by Boveri for his seminal centrosome monograph. PMID:25047623

A phenanthrene derived PARP inhibitor is an extra-centrosomes de-clustering agent exclusively eradicating human cancer cells

PubMed Central

2011-01-01

Background Cells of most human cancers have supernumerary centrosomes. To enable an accurate chromosome segregation and cell division, these cells developed a yet unresolved molecular mechanism, clustering their extra centrosomes at two poles, thereby mimicking mitosis in normal cells. Failure of this bipolar centrosome clustering causes multipolar spindle structures and aberrant chromosomes segregation that prevent normal cell division and lead to 'mitotic catastrophe cell death'. Methods We used cell biology and biochemical methods, including flow cytometry, immunocytochemistry and live confocal imaging. Results We identified a phenanthrene derived PARP inhibitor, known for its activity in neuroprotection under stress conditions, which exclusively eradicated multi-centrosomal human cancer cells (mammary, colon, lung, pancreas, ovarian) while acting as extra-centrosomes de-clustering agent in mitosis. Normal human proliferating cells (endothelial, epithelial and mesenchymal cells) were not impaired. Despite acting as PARP inhibitor, the cytotoxic activity of this molecule in cancer cells was not attributed to PARP inhibition alone. Conclusion We identified a water soluble phenanthridine that exclusively targets the unique dependence of most human cancer cells on their supernumerary centrosomes bi-polar clustering for their survival. This paves the way for a new selective cancer-targeting therapy, efficient in a wide range of human cancers. PMID:21943092

Downregulation of Protein 4.1R impairs centrosome function,bipolar spindle organization and anaphase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Spence, Jeffrey R.; Go, Minjoung M.; Bahmanyar, S.

2006-03-17

Centrosomes nucleate and organize interphase MTs and areinstrumental in the assembly of the mitotic bipolar spindle. Here wereport that two members of the multifunctional protein 4.1 family havedistinct distributions at centrosomes. Protein 4.1R localizes to maturecentrioles whereas 4.1G is a component of the pericentriolar matrixsurrounding centrioles. To selectively probe 4.1R function, we used RNAinterference-mediated depletion of 4.1R without decreasing 4.1Gexpression. 4.1R downregulation reduces MT anchoring and organization atinterphase and impairs centrosome separation during prometaphase.Metaphase chromosomes fail to properly condense/align and spindleorganization is aberrant. Notably 4.1R depletion causes mislocalizationof its binding partner NuMA (Nuclear Mitotic Apparatus Protein),essential for spindle pole focusing,more » and disrupts ninein. Duringanaphase/telophase, 4.1R-depleted cells have lagging chromosomes andaberrant MT bridges. Our data provide functional evidence that 4.1R makescrucial contributions to centrosome integrity and to mitotic spindlestructure enabling mitosis and anaphase to proceed with the coordinatedprecision required to avoid pathological events.« less

Novel Microcephalic Primordial Dwarfism Disorder Associated with Variants in the Centrosomal Protein Ninein

PubMed Central

LaFranchi, Stephen H.; Maliga, Zoltan; Lui, Julian C.; Moon, Jennifer E.; McDeed, Cailin; Henke, Katrin; Zonana, Jonathan; Kingman, Garrett A.; Pers, Tune H.; Baron, Jeffrey; Rosenfeld, Ron G.; Hirschhorn, Joel N.; Harris, Matthew P.; Hwa, Vivian

2012-01-01

Context: Microcephalic primordial dwarfism (MPD) is a rare, severe form of human growth failure in which growth restriction is evident in utero and continues into postnatal life. Single causative gene defects have been identified in a number of patients with MPD, and all involve genes fundamental to cellular processes including centrosome functions. Objective: The objective of the study was to find the genetic etiology of a novel presentation of MPD. Design: The design of the study was whole-exome sequencing performed on two affected sisters in a single family. Molecular and functional studies of a candidate gene were performed using patient-derived primary fibroblasts and a zebrafish morpholino oligonucleotides knockdown model. Patients: Two sisters presented with a novel subtype of MPD, including severe intellectual disabilities. Main Outcome Measures: NIN, encoding Ninein, a centrosomal protein critically involved in asymmetric cell division, was identified as a candidate gene, and functional impacts in fibroblasts and zebrafish were studied. Results: From 34,606 genomic variants, two very rare missense variants in NIN were identified. Both probands were compound heterozygotes. In the zebrafish, ninein knockdown led to specific and novel defects in the specification and morphogenesis of the anterior neuroectoderm, resulting in a deformity of the developing cranium with a small, squared skull highly reminiscent of the human phenotype. Conclusion: We identified a novel clinical subtype of MPD in two sisters who have rare variants in NIN. We show, for the first time, that reduction of ninein function in the developing zebrafish leads to specific deficiencies of brain and skull development, offering a developmental basis for the myriad phenotypes in our patients. PMID:22933543

CEP63 deficiency promotes p53-dependent microcephaly and reveals a role for the centrosome in meiotic recombination.

PubMed

Marjanović, Marko; Sánchez-Huertas, Carlos; Terré, Berta; Gómez, Rocío; Scheel, Jan Frederik; Pacheco, Sarai; Knobel, Philip A; Martínez-Marchal, Ana; Aivio, Suvi; Palenzuela, Lluís; Wolfrum, Uwe; McKinnon, Peter J; Suja, José A; Roig, Ignasi; Costanzo, Vincenzo; Lüders, Jens; Stracker, Travis H

2015-07-09

CEP63 is a centrosomal protein that facilitates centriole duplication and is regulated by the DNA damage response. Mutations in CEP63 cause Seckel syndrome, a human disease characterized by microcephaly and dwarfism. Here we demonstrate that Cep63-deficient mice recapitulate Seckel syndrome pathology. The attrition of neural progenitor cells involves p53-dependent cell death, and brain size is rescued by the deletion of p53. Cell death is not the result of an aberrant DNA damage response but is triggered by centrosome-based mitotic errors. In addition, Cep63 loss severely impairs meiotic recombination, leading to profound male infertility. Cep63-deficient spermatocytes display numerical and structural centrosome aberrations, chromosome entanglements and defective telomere clustering, suggesting that a reduction in centrosome-mediated chromosome movements underlies recombination failure. Our results provide novel insight into the molecular pathology of microcephaly and establish a role for the centrosome in meiotic recombination.

PCM1 is recruited to the centrosome by the cooperative action of DISC1 and BBS4 and is a candidate for psychiatric illness

PubMed Central

Kamiya, Atsushi; Tan, Perciliz L.; Kubo, Ken-ichiro; Engelhard, Caitlin; Ishizuka, Koko; Kubo, Akiharu; Tsukita, Sachiko; Pulver, Ann E.; Nakajima, Kazunori; Cascella, Nicola G.; Katsanis, Nicholas; Sawa, Akira

2009-01-01

Context A role for the centrosome has been suggested in the pathology of major mental illnesses, especially schizophrenia (SZ). Objectives To show that pericentriolar material-1 protein (PCM1) forms a complex at the centrosome with Disrupted-In-Schizophrenia-1 (DISC1) and Bardet-Biedl syndrome-4 protein (BBS4), which provides a crucial pathway for cortical development associated with the pathology of SZ. To identify mutations in the PCM1 gene in a SZ population. Design Interaction of DISC1, PCM1, and BBS proteins was assessed by immunofluorescent staining and co-immunoprecipitation. Effects of PCM1, DISC1, and BBS on centrosomal functions and corticogenesis in vivo were tested by RNAi. PCM1 gene was examined by sequencing 39 exons and flanking splice sites. Setting and Patients Thirty-two probands with SZ from families that had excess allele sharing among affected individuals at 8p22, and 219 Caucasian controls. Main Outcome Measures Protein interaction and recruitment at the centrosome in cells; neuronal migration in the cerebral cortex; variant discovery in PCM1 in SZ patients. Results PCM1 forms a complex with DISC1 and BBS4 through discrete binding domains in each protein. DISC1 and BBS4 are required for targeting PCM1 and other cargo proteins, such as ninein, to the centrosome in a synergistic manner. In the developing cerebral cortex, suppression of PCM1 leads to neuronal migration defects, which are phenocopied by the suppression of either DISC1 or BBS4, and are exacerbated by the concomitant suppression of both. Furtheremore, a nonsense mutation that segregates with schizophrenia-spectrum psychosis is found in one family. Conclusion Our data further support for the role of centrosomal proteins in cortical development and suggest that perturbation of centrosomal function contributes to the development of mental diseases including SZ. PMID:18762586

CEP63 deficiency promotes p53-dependent microcephaly and reveals a role for the centrosome in meiotic recombination

PubMed Central

Marjanović, Marko; Sánchez-Huertas, Carlos; Terré, Berta; Gómez, Rocío; Scheel, Jan Frederik; Pacheco, Sarai; Knobel, Philip A.; Martínez-Marchal, Ana; Aivio, Suvi; Palenzuela, Lluís; Wolfrum, Uwe; McKinnon, Peter J.; Suja, José A.; Roig, Ignasi; Costanzo, Vincenzo; Lüders, Jens; Stracker, Travis H.

2015-01-01

CEP63 is a centrosomal protein that facilitates centriole duplication and is regulated by the DNA damage response. Mutations in CEP63 cause Seckel syndrome, a human disease characterized by microcephaly and dwarfism. Here we demonstrate that Cep63 deficient mice recapitulate Seckel syndrome pathology. The attrition of neural progenitor cells involves p53-dependent cell death and brain size is rescued by the deletion of p53. Cell death is not the result of an aberrant DNA damage response but is triggered by centrosome-based mitotic errors. In addition, Cep63 loss severely impairs meiotic recombination, leading to profound male infertility. Cep63 deficient spermatocytes display numerical and structural centrosome aberrations, chromosome entanglements and defective telomere clustering, suggesting that a reduction in centrosome-mediated chromosome movements underlies recombination failure. Our results provide novel insight into the molecular pathology of microcephaly and establish a role for the centrosome in meiotic recombination. PMID:26158450

Nonequivalence of maternal centrosomes/centrioles in starfish oocytes: selective casting-off of reproductive centrioles into polar bodies.

PubMed

Uetake, Yumi; Kato, Koichi H; Washitani-Nemoto, Setsuko; Nemoto Si, Shin-ichi

2002-07-01

It is believed that in most animals only the paternal centrosome provides the division poles for mitosis in zygotes. This paternal inheritance of the centrosomes depends on the selective loss of the maternal centrosome. In order to understand the mechanism of centrosome inheritance, the behavior of all maternal centrosomes/centrioles was investigated throughout the meiotic and mitotic cycles by using starfish eggs that had polar body (PB) formation suppressed. In starfish oocytes, the centrioles do not duplicate during meiosis II. Hence, each centrosome of the meiosis II spindle has only one centriole, whereas in meiosis I, each has a pair of centrioles. When two pairs of meiosis I centrioles were retained in the cytoplasm of oocytes by complete suppression of PB extrusion, they separated into four single centrioles in meiosis II. However, after completion of the meiotic process, only two of the four single centrioles were found in addition to the pronucleus. When the two single centrioles of a meiosis II spindle were retained in the oocyte cytoplasm by suppressing the extrusion of the second PB, only one centriole was found with the pronucleus after the completion of the meiotic process. When these PB-suppressed eggs were artificially activated to drive the mitotic cycles, all the surviving single centrioles duplicated repeatedly to form pairs of centrioles, which could organize mitotic spindles. These results indicate that the maternal centrioles are not equivalent in their intrinsic stability and reproductive capacity. The centrosomes with the reproductive centrioles are selectively cast off into the PBs, resulting in the mature egg inheriting a nonreproductive centriole, which would degrade shortly after the completion of meiosis. (c) 2002 Elsevier Science (USA).

Centrobin-mediated Regulation of the Centrosomal Protein 4.1-associated Protein (CPAP) Level Limits Centriole Length during Elongation Stage*

PubMed Central

Gudi, Radhika; Haycraft, Courtney J.; Bell, P. Darwin; Li, Zihai; Vasu, Chenthamarakshan

2015-01-01

Microtubule-based centrioles in the centrosome mediate accurate bipolar cell division, spindle orientation, and primary cilia formation. Cellular checkpoints ensure that the centrioles duplicate only once in every cell cycle and achieve precise dimensions, dysregulation of which results in genetic instability and neuro- and ciliopathies. The normal cellular level of centrosomal protein 4.1-associated protein (CPAP), achieved by its degradation at mitosis, is considered as one of the major mechanisms that limits centriole growth at a predetermined length. Here we show that CPAP levels and centriole elongation are regulated by centrobin. Exogenous expression of centrobin causes abnormal elongation of centrioles due to massive accumulation of CPAP in the cell. Conversely, CPAP was undetectable in centrobin-depleted cells, suggesting that it undergoes degradation in the absence of centrobin. Only the reintroduction of full-length centrobin, but not its mutant form that lacks the CPAP binding site, could restore cellular CPAP levels in centrobin-depleted cells, indicating that persistence of CPAP requires its interaction with centrobin. Interestingly, inhibition of the proteasome in centrobin-depleted cells restored the cellular and centriolar CPAP expression, suggesting its ubiquitination and proteasome-mediated degradation when centrobin is absent. Intriguingly, however, centrobin-overexpressing cells also showed proteasome-independent accumulation of ubiquitinated CPAP and abnormal, ubiquitin-positive, elongated centrioles. Overall, our results show that centrobin interacts with ubiquitinated CPAP and prevents its degradation for normal centriole elongation function. Therefore, it appears that loss of centrobin expression destabilizes CPAP and triggers its degradation to restrict the centriole length during biogenesis. PMID:25616662

Epstein-Barr virus thymidine kinase is a centrosomal resident precisely localized to the periphery of centrioles.

PubMed

Gill, Michael B; Kutok, Jeffery L; Fingeroth, Joyce D

2007-06-01

The thymidine kinase (TK) encoded by Epstein-Barr virus (EBV) differs not only from that of the alphaherpesviruses but also from that of the gamma-2 herpesvirus subfamily. Because cellular location is frequently a determinant of regulatory function, to gain insight into additional role(s) of EBV TK and to uncover how the lymphocryptovirus and rhadinovirus enzymes differ, the subcellular localizations of EBV TK and the related cercopithecine herpesvirus-15 TK were investigated. We show that in contrast to those of the other family members, the gamma-1 herpesvirus TKs localize to the centrosome and even more precisely to the periphery of the centriole, tightly encircling the tubulin-rich centrioles in a microtubule-independent fashion. Centrosomal localization is observed in diverse cell types and occurs whether the protein is expressed independently or in the context of lytic EBV infection. Surprisingly, analysis of mutants revealed that the unique N-terminal domain was not critical for targeting to the centrosome, but rather, peptide sequences located C terminal to this domain were key. This is the first herpesvirus protein documented to reside in the centrosome, or microtubule-organizing center, an amembranous organelle that regulates the structural biology of the cell cycle through control of chromosome separation and cytokinesis. More recently, proteasome-mediated degradation of cell cycle regulatory proteins, production and loading of antigenic peptides onto HLA molecules, and transient homing of diverse virion proteins required for entry and/or egress have been shown to be coordinated at the centrosome. Potential implications of centrosomal localization for EBV TK function are discussed.

The C9orf72 repeat expansion disrupts nucleocytoplasmic transport.

PubMed

Zhang, Ke; Donnelly, Christopher J; Haeusler, Aaron R; Grima, Jonathan C; Machamer, James B; Steinwald, Peter; Daley, Elizabeth L; Miller, Sean J; Cunningham, Kathleen M; Vidensky, Svetlana; Gupta, Saksham; Thomas, Michael A; Hong, Ingie; Chiu, Shu-Ling; Huganir, Richard L; Ostrow, Lyle W; Matunis, Michael J; Wang, Jiou; Sattler, Rita; Lloyd, Thomas E; Rothstein, Jeffrey D

2015-09-03

The hexanucleotide repeat expansion (HRE) GGGGCC (G4C2) in C9orf72 is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recent studies support an HRE RNA gain-of-function mechanism of neurotoxicity, and we previously identified protein interactors for the G4C2 RNA including RanGAP1. A candidate-based genetic screen in Drosophila expressing 30 G4C2 repeats identified RanGAP (Drosophila orthologue of human RanGAP1), a key regulator of nucleocytoplasmic transport, as a potent suppressor of neurodegeneration. Enhancing nuclear import or suppressing nuclear export of proteins also suppresses neurodegeneration. RanGAP physically interacts with HRE RNA and is mislocalized in HRE-expressing flies, neurons from C9orf72 ALS patient-derived induced pluripotent stem cells (iPSC-derived neurons), and in C9orf72 ALS patient brain tissue. Nuclear import is impaired as a result of HRE expression in the fly model and in C9orf72 iPSC-derived neurons, and these deficits are rescued by small molecules and antisense oligonucleotides targeting the HRE G-quadruplexes. Nucleocytoplasmic transport defects may be a fundamental pathway for ALS and FTD that is amenable to pharmacotherapeutic intervention.

Centrin 3 is an inhibitor of centrosomal Mps1 and antagonizes centrin 2 function.

PubMed

Sawant, Dwitiya B; Majumder, Shubhra; Perkins, Jennifer L; Yang, Ching-Hui; Eyers, Patrick A; Fisk, Harold A

2015-11-01

Centrins are a family of small, calcium-binding proteins with diverse cellular functions that play an important role in centrosome biology. We previously identified centrin 2 and centrin 3 (Cetn2 and Cetn3) as substrates of the protein kinase Mps1. However, although Mps1 phosphorylation sites control the function of Cetn2 in centriole assembly and promote centriole overproduction, Cetn2 and Cetn3 are not functionally interchangeable, and we show here that Cetn3 is both a biochemical inhibitor of Mps1 catalytic activity and a biological inhibitor of centrosome duplication. In vitro, Cetn3 inhibits Mps1 autophosphorylation at Thr-676, a known site of T-loop autoactivation, and interferes with Mps1-dependent phosphorylation of Cetn2. The cellular overexpression of Cetn3 attenuates the incorporation of Cetn2 into centrioles and centrosome reduplication, whereas depletion of Cetn3 generates extra centrioles. Finally, overexpression of Cetn3 reduces Mps1 Thr-676 phosphorylation at centrosomes, and mimicking Mps1-dependent phosphorylation of Cetn2 bypasses the inhibitory effect of Cetn3, suggesting that the biological effects of Cetn3 are due to the inhibition of Mps1 function at centrosomes. © 2015 Sawant et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

HIV-1 Recruits UPF1 but Excludes UPF2 to Promote Nucleocytoplasmic Export of the Genomic RNA

PubMed Central

Ajamian, Lara; Abel, Karen; Rao, Shringar; Vyboh, Kishanda; García-de-Gracia, Francisco; Soto-Rifo, Ricardo; Kulozik, Andreas E.; Gehring, Niels H.; Mouland, Andrew J.

2015-01-01

Unspliced, genomic HIV-1 RNA (vRNA) is a component of several ribonucleoprotein complexes (RNP) during the viral replication cycle. In earlier work, we demonstrated that the host upframeshift protein 1 (UPF1), a key factor in nonsense-mediated mRNA decay (NMD), colocalized and associated to the viral structural protein Gag during viral egress. In this work, we demonstrate a new function for UPF1 in the regulation of vRNA nuclear export. We establish that the nucleocytoplasmic shuttling of UPF1 is required for this function and demonstrate that UPF1 exists in two essential viral RNPs during the late phase of HIV-1 replication: the first, in a nuclear export RNP that contains Rev, CRM1, DDX3 and the nucleoporin p62, and the second, which excludes these nuclear export markers but contains Gag in the cytoplasm. Interestingly, we observed that both UPF2 and the long isoform of UPF3a, UPF3aL, but not the shorter isoforms UPF3aS and UPF3b, are excluded from the UPF1-Rev-CRM1-DDX3 complex as they are negative regulators of vRNA nuclear export. In silico protein-protein docking analyses suggest that Rev binds UPF1 in a region that overlaps the UPF2 binding site, thus explaining the exclusion of this negative regulatory factor by HIV-1 that is necessary for vRNA trafficking. This work uncovers a novel and unique regulatory circuit involving several UPF proteins that ultimately regulate vRNA nuclear export and trafficking. PMID:26492277

BRCA1 interaction of centrosomal protein Nlp is required for successful mitotic progression.

PubMed

Jin, Shunqian; Gao, Hua; Mazzacurati, Lucia; Wang, Yang; Fan, Wenhong; Chen, Qiang; Yu, Wei; Wang, Mingrong; Zhu, Xueliang; Zhang, Chuanmao; Zhan, Qimin

2009-08-21

Breast cancer susceptibility gene BRCA1 is implicated in the control of mitotic progression, although the underlying mechanism(s) remains to be further defined. Deficiency of BRCA1 function leads to disrupted mitotic machinery and genomic instability. Here, we show that BRCA1 physically interacts and colocalizes with Nlp, an important molecule involved in centrosome maturation and spindle formation. Interestingly, Nlp centrosomal localization and its protein stability are regulated by normal cellular BRCA1 function because cells containing BRCA1 mutations or silenced for endogenous BRCA1 exhibit disrupted Nlp colocalization to centrosomes and enhanced Nlp degradation. Its is likely that the BRCA1 regulation of Nlp stability involves Plk1 suppression. Inhibition of endogenous Nlp via the small interfering RNA approach results in aberrant spindle formation, aborted chromosomal segregation, and aneuploidy, which mimic the phenotypes of disrupted BRCA1. Thus, BRCA1 interaction of Nlp might be required for the successful mitotic progression, and abnormalities of Nlp lead to genomic instability.

BRCA1 and FancJ cooperatively promote interstrand crosslinker induced centrosome amplification through the activation of polo-like kinase 1

PubMed Central

Zou, Jianqiu; Zhang, Deli; Qin, Guang; Chen, Xiangming; Wang, Hongmin; Zhang, Dong

2014-01-01

DNA damage response (DDR) and the centrosome cycle are 2 of the most critical cellular processes affecting the genome stability in animal cells. Yet the cross-talks between DDR and the centrosome are poorly understood. Here we showed that deficiency of the breast cancer 1, early onset gene (BRCA1) induces centrosome amplification in non-stressed cells as previously reported while attenuating DNA damage-induced centrosome amplification (DDICA) in cells experiencing prolonged genotoxic stress. Mechanistically, the function of BRCA1 in promoting DDICA is through binding and recruiting polo-like kinase 1 (PLK1) to the centrosome. In a recent study, we showed that FancJ also suppresses centrosome amplification in non-stressed cells while promoting DDICA in both hydroxyurea and mitomycin C treated cells. FancJ is a key component of the BRCA1 B-complex. Here, we further demonstrated that, in coordination with BRCA1, FancJ promotes DDICA by recruiting both BRCA1 and PLK1 to the centrosome in the DNA damaged cells. Thus, we have uncovered a novel role of BRCA1 and FancJ in the regulation of DDICA. Dysregulation of DDR or centrosome cycle leads to aneuploidy, which is frequently seen in both solid and hematological cancers. BRCA1 and FancJ are known tumor suppressors and have well-recognized functions in DNA damage checkpoint and DNA repair. Together with our recent findings, we demonstrated here that BRCA1 and FancJ also play an important role in centrosome cycle especially in DDICA. DDICA is thought to be an alternative fail-safe mechanism to prevent cells experiencing severe DNA damage from becoming carcinogenic. Therefore, BRCA1 and FancJ are potential liaisons linking early DDR with the DDICA. We propose that together with their functions in DDR, the role of BRCA1 and FancJ in the activation of DDICA is also crucial for their tumor suppression functions in vivo. PMID:25483079

Centrosomal Latency of Incoming Foamy Viruses in Resting Cells

PubMed Central

Giron, Marie Lou; Roingeard, Philippe; Clave, Emmanuel; Tobaly-Tapiero, Joelle; Bittoun, Patricia; Toubert, Antoine; de Thé, Hugues; Saïb, Ali

2007-01-01

Completion of early stages of retrovirus infection depends on the cell cycle. While gammaretroviruses require mitosis for proviral integration, lentiviruses are able to replicate in post-mitotic non-dividing cells. Resting cells such as naive resting T lymphocytes from peripheral blood cannot be productively infected by retroviruses, including lentiviruses, but the molecular basis of this restriction remains poorly understood. We demonstrate that in G0 resting cells (primary fibroblasts or peripheral T cells), incoming foamy retroviruses accumulate in close proximity to the centrosome, where they lie as structured and assembled capsids for several weeks. Under these settings, virus uncoating is impaired, but upon cell stimulation, Gag proteolysis and capsid disassembly occur, which allows viral infection to proceed. The data imply that foamy virus uncoating is the rate-limiting step for productive infection of primary G0 cells. Incoming foamy retroviruses can stably persist at the centrosome, awaiting cell stimulation to initiate capsid cleavage, nuclear import, and viral gene expression. PMID:17530924

BRCA1 Interaction of Centrosomal Protein Nlp Is Required for Successful Mitotic Progression*♦

PubMed Central

Jin, Shunqian; Gao, Hua; Mazzacurati, Lucia; Wang, Yang; Fan, Wenhong; Chen, Qiang; Yu, Wei; Wang, Mingrong; Zhu, Xueliang; Zhang, Chuanmao; Zhan, Qimin

2009-01-01

Breast cancer susceptibility gene BRCA1 is implicated in the control of mitotic progression, although the underlying mechanism(s) remains to be further defined. Deficiency of BRCA1 function leads to disrupted mitotic machinery and genomic instability. Here, we show that BRCA1 physically interacts and colocalizes with Nlp, an important molecule involved in centrosome maturation and spindle formation. Interestingly, Nlp centrosomal localization and its protein stability are regulated by normal cellular BRCA1 function because cells containing BRCA1 mutations or silenced for endogenous BRCA1 exhibit disrupted Nlp colocalization to centrosomes and enhanced Nlp degradation. Its is likely that the BRCA1 regulation of Nlp stability involves Plk1 suppression. Inhibition of endogenous Nlp via the small interfering RNA approach results in aberrant spindle formation, aborted chromosomal segregation, and aneuploidy, which mimic the phenotypes of disrupted BRCA1. Thus, BRCA1 interaction of Nlp might be required for the successful mitotic progression, and abnormalities of Nlp lead to genomic instability. PMID:19509300

Mitosis-specific phosphorylation of amyloid precursor protein at Threonine 668 leads to its altered processing and association with centrosomes

PubMed Central

2011-01-01

Background Atypical expression of cell cycle regulatory proteins has been implicated in Alzheimer's disease (AD), but the molecular mechanisms by which they induce neurodegeneration are not well understood. We examined transgenic mice expressing human amyloid precursor protein (APP) and presenilin 1 (PS1) for changes in cell cycle regulatory proteins to determine whether there is a correlation between cell cycle activation and pathology development in AD. Results Our studies in the AD transgenic mice show significantly higher levels of cyclin E, cyclin D1, E2F1, and P-cdc2 in the cells in the vicinity of the plaques where maximum levels of Threonine 668 (Thr668)-phosphorylated APP accumulation was observed. This suggests that the cell cycle regulatory proteins might be influencing plaque pathology by affecting APP phosphorylation. Using neuroglioma cells overexpressing APP we demonstrate that phosphorylation of APP at Thr668 is mitosis-specific. Cells undergoing mitosis show altered cellular distribution and localization of P-APP at the centrosomes. Also, Thr668 phosphorylation in mitosis correlates with increased processing of APP to generate Aβ and the C-terminal fragment of APP, which is prevented by pharmacological inhibitors of the G1/S transition. Conclusions The data presented here suggests that cell cycle-dependent phosphorylation of APP may affect its normal cellular function. For example, association of P-APP with the centrosome may affect spindle assembly and cell cycle progression, further contributing to the development of pathology in AD. The experiments with G1/S inhibitors suggest that cell cycle inhibition may impede the development of Alzheimer's pathology by suppressing modification of βAPP, and thus may represent a novel approach to AD treatment. Finally, the cell cycle regulated phosphorylation and processing of APP into Aβ and the C-terminal fragment suggest that these proteins may have a normal function during mitosis. PMID:22112898

Activity and subcellular compartmentalization of peroxisome proliferator-activated receptor alpha are altered by the centrosome-associated protein CAP350.

PubMed

Patel, Hansa; Truant, Ray; Rachubinski, Richard A; Capone, John P

2005-01-01

Peroxisome proliferator-activated nuclear hormone receptors (PPAR) are ligand-activated transcription factors that play pivotal roles in governing metabolic homeostasis and cell growth. PPARs are primarily in the nucleus but, under certain circumstances, can be found in the cytoplasm. We show here that PPAR(alpha) interacts with the centrosome-associated protein CAP350. CAP350 also interacts with PPAR(delta), PPAR(gamma) and liver-X-receptor alpha, but not with the 9-cis retinoic acid receptor, RXR(alpha). Immunofluorescence analysis indicated that PPAR(alpha) is diffusely distributed in the nucleus and excluded from the cytoplasm. However, in the presence of coexpressed CAP350, PPAR(alpha) colocalizes with CAP350 to discrete nuclear foci and to the centrosome, perinuclear region and intermediate filaments. In contrast, the subcellular distribution of RXR(alpha) or of thyroid hormone receptor alpha was not altered by coexpression of CAP350. An amino-terminal fragment of CAP350 was localized exclusively to nuclear foci and was sufficient to recruit PPAR(alpha) to these sites. Mutation of the single putative nuclear hormone receptor interacting signature motif LXXLL present in this fragment had no effect on its subnuclear localization but abrogated recruitment of PPAR(alpha) to nuclear foci. Surprisingly, mutation of the LXXLL motif in this CAP350 subfragment did not prevent its binding to PPAR(alpha) in vitro, suggesting that this motif serves some function other than PPAR(alpha) binding in recruiting PPAR(alpha) to nuclear spots. CAP350 inhibited PPAR(alpha)-mediated transactivation in an LXXLL-dependent manner, suggesting that CAP350 represses PPAR(alpha) function. Our findings implicate CAP350 in a dynamic process that recruits PPAR(alpha) to discrete nuclear and cytoplasmic compartments and suggest that altered intracellular compartmentalization represents a regulatory process that modulates PPAR function.

Increased centrosome number in BRCA-related breast cancer specimens determined by immunofluorescence analysis.

PubMed

Watanabe, Gou; Chiba, Natsuko; Nomizu, Tadashi; Furuta, Akihiko; Sato, Kaolu; Miyashita, Minoru; Tada, Hiroshi; Suzuki, Akihiko; Ohuchi, Noriaki; Ishida, Takanori

2018-06-01

BRCA-related breast carcinoma can be prevented through prophylactic surgery and an intensive follow-up regimen. However, BRCA genetic tests cannot be routinely performed, and some BRCA mutations could not be defined as deleterious mutations or normal variants. Therefore, an easy functional assay of BRCA will be useful to evaluate BRCA status. As it has been reported that BRCA functions in the regulation of centrosome number, we focused on centrosome number in cancer tissues. Here, 70 breast cancer specimens with known BRCA status were analyzed using immunofluorescence of γ-tubulin (a marker of centrosome) foci. The number of foci per cell was higher in cases with BRCA mutation compared to wild-type cases, that is, 1.9 (95% confidence interval [CI], 1.5-2.3) vs 0.5 (95% CI, 0.2-0.8) (P < .001). Specifically, foci numbers per cell in BRCA1 and BRCA2 mutation cases were 1.2 (95% CI, 0.6-1.8) and 2.2 (95% CI, 1.7-2.6), respectively, both higher than those in wild-type cases (P = .042 and P < .0001, respectively). The predictive value of γ-tubulin foci as determined by area under the curve (AUC = 0.86) for BRCA status was superior to BRCAPRO (AUC = 0.69), Myriad Table (AUC = 0.61), and KOHBRA BRCA risk calculator (AUC = 0.65) pretest values. The use of γ-tubulin foci to predict BRCA status had sensitivity = 83% (19/23), specificity = 89% (42/47), and positive predictive value = 77% (20/26). Thus, γ-tubulin immunofluorescence, a functional assessment of BRCA, can be used as a new prospective test of BRCA status. © 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

Cdk1 Phosphorylates Drosophila Sas-4 to Recruit Polo to Daughter Centrioles and Convert Them to Centrosomes.

PubMed

Novak, Zsofia A; Wainman, Alan; Gartenmann, Lisa; Raff, Jordan W

2016-06-20

Centrosomes and cilia are organized by a centriole pair comprising an older mother and a younger daughter. Centriole numbers are tightly regulated, and daughter centrioles (which assemble in S phase) cannot themselves duplicate or organize centrosomes until they have passed through mitosis. It is unclear how this mitotic "centriole conversion" is regulated, but it requires Plk1/Polo kinase. Here we show that in flies, Cdk1 phosphorylates the conserved centriole protein Sas-4 during mitosis. This creates a Polo-docking site that helps recruit Polo to daughter centrioles and is required for the subsequent recruitment of Asterless (Asl), a protein essential for centriole duplication and mitotic centrosome assembly. Point mutations in Sas-4 that prevent Cdk1 phosphorylation or Polo docking do not block centriole disengagement during mitosis, but block efficient centriole conversion and lead to embryonic lethality. These observations can explain why daughter centrioles have to pass through mitosis before they can duplicate and organize a centrosome. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

HIV-1 Recruits UPF1 but Excludes UPF2 to Promote Nucleocytoplasmic Export of the Genomic RNA.

PubMed

Ajamian, Lara; Abel, Karen; Rao, Shringar; Vyboh, Kishanda; García-de-Gracia, Francisco; Soto-Rifo, Ricardo; Kulozik, Andreas E; Gehring, Niels H; Mouland, Andrew J

2015-10-20

Unspliced, genomic HIV-1 RNA (vRNA) is a component of several ribonucleoprotein complexes (RNP) during the viral replication cycle. In earlier work, we demonstrated that the host upframeshift protein 1 (UPF1), a key factor in nonsense-mediated mRNA decay (NMD), colocalized and associated to the viral structural protein Gag during viral egress. In this work, we demonstrate a new function for UPF1 in the regulation of vRNA nuclear export. OPEN ACCESS Biomolecules 2015, 5 2809 We establish that the nucleocytoplasmic shuttling of UPF1 is required for this function and demonstrate that UPF1 exists in two essential viral RNPs during the late phase of HIV-1 replication: the first, in a nuclear export RNP that contains Rev, CRM1, DDX3 and the nucleoporin p62, and the second, which excludes these nuclear export markers but contains Gag in the cytoplasm. Interestingly, we observed that both UPF2 and the long isoform of UPF3a, UPF3aL, but not the shorter isoforms UPF3aS and UPF3b, are excluded from the UPF1-Rev-CRM1-DDX3 complex as they are negative regulators of vRNA nuclear export. In silico protein-protein docking analyses suggest that Rev binds UPF1 in a region that overlaps the UPF2 binding site, thus explaining the exclusion of this negative regulatory factor by HIV-1 that is necessary for vRNA trafficking. This work uncovers a novel and unique regulatory circuit involving several UPF proteins that ultimately regulate vRNA nuclear export and trafficking.

Centrosome and microtubule instability in aging Drosophila cells

NASA Technical Reports Server (NTRS)

Schatten, H.; Chakrabarti, A.; Hedrick, J.

1999-01-01

Several cytoskeletal changes are associated with aging which includes alterations in muscle structure leading to muscular atrophy, and weakening of the microtubule network which affects cellular secretion and maintenance of cell shape. Weakening of the microtubule network during meiosis in aging oocytes can result in aneuploidy or trisomic zygotes with increasing maternal age. Imbalances of cytoskeletal organization can lead to disease such as Alzheimer's, muscular disorders, and cancer. Because many cytoskeletal diseases are related to age we investigated the effects of aging on microtubule organization in cell cultures of the Drosophila cell model system (Schneider S-1 and Kc23 cell lines). This cell model is increasingly being used as an alternative system to mammalian cell cultures. Drosophila cells are amenable to genetic manipulations and can be used to identify and manipulate genes which are involved in the aging processes. Immunofluorescence, scanning, and transmission electron microscopy were employed for the analysis of microtubule organizing centers (centrosomes) and microtubules at various times after subculturing cells in fresh medium. Our results reveal that centrosomes and the microtubule network becomes significantly affected in aging cells after 5 days of subculture. At 5-14 days of subculture, 1% abnormal out of 3% mitoses were noted which were clearly distinguishable from freshly subcultured control cells in which 3% of cells undergo normal mitosis with bipolar configurations. Microtubules are also affected in the midbody during cell division. The midbody in aging cells becomes up to 10 times longer when compared with midbodies in freshly subcultured cells. During interphase, microtubules are often disrupted and disorganized, which may indicate improper function related to transport of cell organelles along microtubules. These results are likely to help explain some cytoskeletal disorders and diseases related to aging.

Discovery of a nucleocytoplasmic O-mannose glycoproteome in yeast

PubMed Central

Halim, Adnan; Larsen, Ida Signe Bohse; Neubert, Patrick; Joshi, Hiren Jitendra; Petersen, Bent Larsen; Vakhrushev, Sergey Y.; Strahl, Sabine; Clausen, Henrik

2015-01-01

Dynamic cycling of N-Acetylglucosamine (GlcNAc) on serine and threonine residues (O-GlcNAcylation) is an essential process in all eukaryotic cells except yeast, including Saccharomyces cerevisiae and Schizosaccharomyces pombe. O-GlcNAcylation modulates signaling and cellular processes in an intricate interplay with protein phosphorylation and serves as a key sensor of nutrients by linking the hexosamine biosynthetic pathway to cellular signaling. A longstanding conundrum has been how yeast survives without O-GlcNAcylation in light of its similar phosphorylation signaling system. We previously developed a sensitive lectin enrichment and mass spectrometry workflow for identification of the human O-linked mannose (O-Man) glycoproteome and used this to identify a pleothora of O-Man glycoproteins in human cell lines including the large family of cadherins and protocadherins. Here, we applied the workflow to yeast with the aim to characterize the yeast O-Man glycoproteome, and in doing so, we discovered hitherto unknown O-Man glycosites on nuclear, cytoplasmic, and mitochondrial proteins in S. cerevisiae and S. pombe. Such O-Man glycoproteins were not found in our analysis of human cell lines. However, the type of yeast O-Man nucleocytoplasmic proteins and the localization of identified O-Man residues mirror that of the O-GlcNAc glycoproteome found in other eukaryotic cells, indicating that the two different types of O-glycosylations serve the same important biological functions. The discovery opens for exploration of the enzymatic machinery that is predicted to regulate the nucleocytoplasmic O-Man glycosylations. It is likely that manipulation of this type of O-Man glycosylation will have wide applications for yeast bioprocessing. PMID:26644575

The Stil protein regulates centrosome integrity and mitosis through suppression of Chfr

PubMed Central

Castiel, Asher; Danieli, Michal Mark; David, Ahuvit; Moshkovitz, Sharon; Aplan, Peter D.; Kirsch, Ilan R.; Brandeis, Michael; Krämer, Alwin; Izraeli, Shai

2011-01-01

Stil (Sil, SCL/TAL1 interrupting locus) is a cytosolic and centrosomal protein expressed in proliferating cells that is required for mouse and zebrafish neural development and is mutated in familial microcephaly. Recently the Drosophila melanogaster ortholog of Stil was found to be important for centriole duplication. Consistent with this finding, we report here that mouse embryonic fibroblasts lacking Stil are characterized by slow growth, low mitotic index and absence of clear centrosomes. We hypothesized that Stil regulates mitosis through the tumor suppressor Chfr, an E3 ligase that blocks mitotic entry in response to mitotic stress. Mouse fibroblasts lacking Stil by genomic or RNA interference approaches, as well as E9.5 Stil−/− embryos, express high levels of the Chfr protein and reduced levels of the Chfr substrate Plk1. Exogenous expression of Stil, knockdown of Chfr or overexpression of Plk1 reverse the abnormal mitotic phenotypes of fibroblasts lacking Stil. We further demonstrate that Stil increases Chfr auto-ubiquitination and reduces its protein stability. Thus, Stil is required for centrosome organization, entry into mitosis and cell proliferation, and these functions are at least partially mediated by Chfr and its targets. This is the first identification of a negative regulator of the Chfr mitotic checkpoint. PMID:21245198

Modular elements of the TPR domain in the Mps1 N terminus differentially target Mps1 to the centrosome and kinetochore

PubMed Central

Marquardt, Joseph R.; Perkins, Jennifer L.; Beuoy, Kyle J.; Fisk, Harold A.

2016-01-01

Faithful segregation of chromosomes to two daughter cells is regulated by the formation of a bipolar mitotic spindle and the spindle assembly checkpoint, ensuring proper spindle function. Here we show that the proper localization of the kinase Mps1 (monopolar spindle 1) is critical to both these processes. Separate elements in the Mps1 N-terminal extension (NTE) and tetratricopeptide repeat (TPR) domains govern localization to either the kinetochore or the centrosome. The third TPR (TPR3) and the TPR-capping helix (C-helix) are each sufficient to target Mps1 to the centrosome. TPR3 binds to voltage-dependent anion channel 3, but although this is sufficient for centrosome targeting of Mps1, it is not necessary because of the presence of the C-helix. A version of Mps1 lacking both elements cannot localize to or function at the centrosome, but maintains kinetochore localization and spindle assembly checkpoint function, indicating that TPR3 and the C-helix define a bipartite localization determinant that is both necessary and sufficient to target Mps1 to the centrosome but dispensable for kinetochore targeting. In contrast, elements required for kinetochore targeting (the NTE and first two TPRs) are dispensable for centrosomal localization and function. These data are consistent with a separation of Mps1 function based on localization determinants within the N terminus. PMID:27339139

Modular elements of the TPR domain in the Mps1 N terminus differentially target Mps1 to the centrosome and kinetochore.

PubMed

Marquardt, Joseph R; Perkins, Jennifer L; Beuoy, Kyle J; Fisk, Harold A

2016-07-12

Faithful segregation of chromosomes to two daughter cells is regulated by the formation of a bipolar mitotic spindle and the spindle assembly checkpoint, ensuring proper spindle function. Here we show that the proper localization of the kinase Mps1 (monopolar spindle 1) is critical to both these processes. Separate elements in the Mps1 N-terminal extension (NTE) and tetratricopeptide repeat (TPR) domains govern localization to either the kinetochore or the centrosome. The third TPR (TPR3) and the TPR-capping helix (C-helix) are each sufficient to target Mps1 to the centrosome. TPR3 binds to voltage-dependent anion channel 3, but although this is sufficient for centrosome targeting of Mps1, it is not necessary because of the presence of the C-helix. A version of Mps1 lacking both elements cannot localize to or function at the centrosome, but maintains kinetochore localization and spindle assembly checkpoint function, indicating that TPR3 and the C-helix define a bipartite localization determinant that is both necessary and sufficient to target Mps1 to the centrosome but dispensable for kinetochore targeting. In contrast, elements required for kinetochore targeting (the NTE and first two TPRs) are dispensable for centrosomal localization and function. These data are consistent with a separation of Mps1 function based on localization determinants within the N terminus.

Transactivation activity and nucleocytoplasmic transport of β-catenin are independently regulated by its C-terminal end.

PubMed

Maturana, J L; Niechi, I; Silva, E; Huerta, H; Cataldo, R; Härtel, S; Barros, L F; Galindo, M; Tapia, J C

2015-11-15

The key protein in the canonical Wnt pathway is β-catenin, which is phosphorylated both in absence and presence of Wnt signals by different kinases. Upon activation in the cytoplasm, β-catenin can enter into the nucleus to transactivate target gene expression, many of which are cancer-related genes. The mechanism governing β-catenin's nucleocytoplasmic transport has been recently unvealed, although phosphorylation at its C-terminal end and its functional consequences are not completely understood. Serine 646 of β-catenin is a putative CK2 phosphorylation site and lies in a region which has been proposed to be important for its nucleocytoplasmic transport and transactivation activity. This residue was mutated to aspartic acid mimicking CK2-phosphorylation and its effects on β-catenin activity as well as localization were explored. β-Catenin S6464D did not show significant differences in both transcriptional activity and nuclear localization compared to the wild-type form, but displayed a characteristic granular nuclear pattern. Three-dimensional models of nuclei were constructed which showed differences in number and volume of granules, being those from β-catenin S646D more and smaller than the wild-type form. FRAP microscopy was used to compare nuclear export of both proteins which showed a slightly higher but not significant retention of β-catenin S646D. Altogether, these results show that C-terminal phosphorylation of β-catenin seems to be related with its nucleocytoplasmic transport but not transactivation activity. Copyright © 2015 Elsevier B.V. All rights reserved.

KENNEDY SPACE CENTER, FLA. -- From left, United Space Alliance (USA) Deputy Space Shuttle Program Manager of Operations Loren Shriver, USA Associate Program Manager of Ground Operations Andy Allen, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, and USA Vice President and Space Shuttle Program Manager Howard DeCastro examine a tile used in the Shuttle's Thermal Protection System (TPS) in KSC's TPS Facility. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, United Space Alliance (USA) Deputy Space Shuttle Program Manager of Operations Loren Shriver, USA Associate Program Manager of Ground Operations Andy Allen, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, and USA Vice President and Space Shuttle Program Manager Howard DeCastro examine a tile used in the Shuttle's Thermal Protection System (TPS) in KSC's TPS Facility. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

The presence of centrioles and centrosomes in ovarian mature cystic teratoma cells suggests human parthenotes developed in vitro can differentiate into mature cells without a sperm centriole

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Bo Yon, E-mail: boyonlee@gmail.com; Shim, Sang Woo; Kim, Young Sun

Highlights: Black-Right-Pointing-Pointer The sperm centriole is the progenitor of centrosomes in all somatic cells. Black-Right-Pointing-Pointer Centrioles and centrosomes exist in parthenogenetic ovarian teratoma cells. Black-Right-Pointing-Pointer Without a sperm centriole, parthenogenetic oocytes produce centrioles and centrosomes. Black-Right-Pointing-Pointer Parthenogenetic human oocytes can develop and differentiate into mature cells. -- Abstract: In most animals, somatic cell centrosomes are inherited from the centriole of the fertilizing spermatozoa. The oocyte centriole degenerates during oogenesis, and completely disappears in metaphase II. Therefore, the embryos generated by in vitro parthenogenesis are supposed to develop without any centrioles. Exceptional acentriolar and/or acentrosomal developments are possible in mice andmore » in some experimental cells; however, in most animals, the full developmental potential of parthenogenetic cells in vitro and the fate of their centrioles/centrosomes are not clearly understood. To predict the future of in vitro human parthenogenesis, we explored the centrioles/centrosomes in ovarian mature cystic teratoma cells by immunofluorescent staining and transmission electron microscopy. We confirmed the presence of centrioles and centrosomes in these well-known parthenogenetic ovarian tumor cells. Our findings clearly demonstrate that, even without a sperm centriole, parthenotes that develop from activated oocytes can produce their own centrioles/centrosomes, and can even develop into the well-differentiated mature tissue.« less

Centrosomal Nlp is an oncogenic protein that is gene-amplified in human tumors and causes spontaneous tumorigenesis in transgenic mice.

PubMed

Shao, Shujuan; Liu, Rong; Wang, Yang; Song, Yongmei; Zuo, Lihui; Xue, Liyan; Lu, Ning; Hou, Ning; Wang, Mingrong; Yang, Xiao; Zhan, Qimin

2010-02-01

Disruption of mitotic events contributes greatly to genomic instability and results in mutator phenotypes. Indeed, abnormalities of mitotic components are closely associated with malignant transformation and tumorigenesis. Here we show that ninein-like protein (Nlp), a recently identified BRCA1-associated centrosomal protein involved in microtubule nucleation and spindle formation, is an oncogenic protein. Nlp was found to be overexpressed in approximately 80% of human breast and lung carcinomas analyzed. In human lung cancers, this deregulated expression was associated with NLP gene amplification. Further analysis revealed that Nlp exhibited strong oncogenic properties; for example, it conferred to NIH3T3 rodent fibroblasts the capacity for anchorage-independent growth in vitro and tumor formation in nude mice. Consistent with these data, transgenic mice overexpressing Nlp displayed spontaneous tumorigenesis in the breast, ovary, and testicle within 60 weeks. In addition, Nlp overexpression induced more rapid onset of radiation-induced lymphoma. Furthermore, mouse embryonic fibroblasts (MEFs) derived from Nlp transgenic mice showed centrosome amplification, suggesting that Nlp overexpression mimics BRCA1 loss. These findings demonstrate that Nlp abnormalities may contribute to genomic instability and tumorigenesis and suggest that Nlp might serve as a potential biomarker for clinical diagnosis and therapeutic target.

NASA Space Shuttle Program: Shuttle Environmental Assurance (SEA) Initiative

NASA Technical Reports Server (NTRS)

Glover, Steve E.; McCool, Alex (Technical Monitor)

2002-01-01

The first Space Shuttle flight was in 1981 and the fleet was originally expected to be replaced with a new generation vehicle in the early 21st century. Space Shuttle Program (SSP) elements proactively address environmental and obsolescence concerns and continue to improve safety and supportability. The SSP manager created the Shuttle Environmental Assurance (SEA) Initiative in 2000. SEA is to provide an integrated approach for the SSP to promote environmental excellence, proactively manage materials obsolescence, and optimize associated resources.

Purinergic A2b Receptor Activation by Extracellular Cues Affects Positioning of the Centrosome and Nucleus and Causes Reduced Cell Migration*

PubMed Central

Ou, Young; Chan, Gordon; Zuo, Jeremy; Rattner, Jerome B.; van der Hoorn, Frans A.

2016-01-01

The tight, relative positioning of the nucleus and centrosome in mammalian cells is important for the regulation of cell migration. Under pathophysiological conditions, the purinergic A2b receptor can regulate cell motility, but the underlying mechanism remains unknown. Expression of A2b, normally low, is increased in tissues experiencing adverse physiological conditions, including hypoxia and inflammation. ATP is released from such cells. We investigated whether extracellular cues can regulate centrosome-nucleus positioning and cell migration. We discovered that hypoxia as well as extracellular ATP cause a reversible increase in the distance between the centrosome and nucleus and reduced cell motility. We uncovered the underlying pathway: both treatments act through the A2b receptor and specifically activate the Epac1/RapGef3 pathway. We show that cells lacking A2b do not respond in this manner to hypoxia or ATP but transfection of A2b restores this response, that Epac1 is critically involved, and that Rap1B is important for the relative positioning of the centrosome and nucleus. Our results represent, to our knowledge, the first report demonstrating that pathophysiological conditions can impact the distance between the centrosome and nucleus. Furthermore, we identify the A2b receptor as a central player in this process. PMID:27226580

Poor Centrosomal Function of Cat Testicular Spermatozoa Impairs Embryo Development In Vitro after Intracytoplasmic Sperm Injection1

PubMed Central

Comizzoli, Pierre; Wildt, David E.; Pukazhenthi, Budhan S.

2007-01-01

In the domestic cat, morula-blastocyst formation in vitro is compromised after intracytoplasmic sperm injection (ICSI) with testicular compared to ejaculated spermatozoa. The aim of this study was to determine the cellular basis of the lower developmental potential of testicular spermatozoa. Specifically, we examined the influence of sperm DNA fragmentation (evaluated by TUNEL assay) and centrosomal function (assessed by sperm aster formation after ICSI) on first-cleavage timing, developmental rate, and morula-blastocyst formation. Because the incidences of DNA fragmentation were not different between testicular and ejaculated sperm suspensions, DNA integrity was not the origin of the reduced developmental potential of testicular spermatozoa. After ICSI, proportions of fertilized and cleaved oocytes were similar and not influenced by sperm source. However, observations made at 5 h post-activation clearly demonstrated that 1) zygotes generally contained a large sperm aster after ICSI with ejaculated spermatozoa, a phenomenon never observed with testicular spermatozoa, and 2) proportions of zygotes with short or absent sperm asters were higher after ICSI with testicular spermatozoa than using ejaculated spermatozoa. The poor pattern of aster formation arose from the testicular sperm centrosome, which contributed to a delayed first cleavage, a slower developmental rate, and a reduced formation of morulae and blastocysts compared to ejaculated spermatozoa. When a testicular sperm centrosome was replaced by a centrosome from an ejaculated spermatozoon, kinetics of first cell cycle as well as embryo development quality significantly improved and were comparable to data from ejaculated spermatozoa. Results demonstrate for the first time in mammals that maturity of the cat sperm centrosome (likely via epididymal transit) contributes to an enhanced ability of the spermatozoon to produce embryos that develop normally to the morula and blastocyst stages. PMID:16687647

A centrosomal protein FOR20 regulates microtubule assembly dynamics and plays a role in cell migration.

PubMed

Srivastava, Shalini; Panda, Dulal

2017-08-10

Here, we report that a centrosomal protein FOR20 [FOP (FGFR1 (fibroblast growth factor receptor 1) oncogene protein)-like protein of molecular mass of 20 kDa; also named as C16orf63, FLJ31153 or PHSECRG2] can regulate the assembly and stability of microtubules. Both FOR20 IgG antibody and GST (glutathione S -transferase)-tagged FOR20 could precipitate tubulin from the HeLa cell extract, indicating a possible interaction between FOR20 and tubulin. FOR20 was also detected in goat brain tissue extract and it cycled with microtubule-associated proteins. Furthermore, FOR20 bound to purified tubulin and inhibited the assembly of tubulin in vitro. The overexpression of FOR20 depolymerized interphase microtubules and the depletion of FOR20 prevented nocodazole-induced depolymerization of microtubules in HeLa cells. In addition, the depletion of FOR20 suppressed the dynamics of individual microtubules in live HeLa cells. FOR20-depleted MDA-MB-231 cells displayed zigzag motion and migrated at a slower rate than the control cells, indicating that FOR20 plays a role in directed cell migration. The results suggested that the centrosomal protein FOR20 is a new member of the microtubule-associated protein family and that it regulates the assembly and dynamics of microtubules. © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

STED nanoscopy of the centrosome linker reveals a CEP68-organized, periodic rootletin network anchored to a C-Nap1 ring at centrioles.

PubMed

Vlijm, Rifka; Li, Xue; Panic, Marko; Rüthnick, Diana; Hata, Shoji; Herrmannsdörfer, Frank; Kuner, Thomas; Heilemann, Mike; Engelhardt, Johann; Hell, Stefan W; Schiebel, Elmar

2018-03-06

The centrosome linker proteins C-Nap1, rootletin, and CEP68 connect the two centrosomes of a cell during interphase into one microtubule-organizing center. This coupling is important for cell migration, cilia formation, and timing of mitotic spindle formation. Very little is known about the structure of the centrosome linker. Here, we used stimulated emission depletion (STED) microscopy to show that each C-Nap1 ring at the proximal end of the two centrioles organizes a rootletin ring and, in addition, multiple rootletin/CEP68 fibers. Rootletin/CEP68 fibers originating from the two centrosomes form a web-like, interdigitating network, explaining the flexible nature of the centrosome linker. The rootletin/CEP68 filaments are repetitive and highly ordered. Staggered rootletin molecules (N-to-N and C-to-C) within the filaments are 75 nm apart. Rootletin binds CEP68 via its C-terminal spectrin repeat-containing region in 75-nm intervals. The N-to-C distance of two rootletin molecules is ∼35 to 40 nm, leading to an estimated minimal rootletin length of ∼110 nm. CEP68 is important in forming rootletin filaments that branch off centrioles and to modulate the thickness of rootletin fibers. Thus, the centrosome linker consists of a vast network of repeating rootletin units with C-Nap1 as ring organizer and CEP68 as filament modulator. Copyright © 2018 the Author(s). Published by PNAS.

Purinergic A2b Receptor Activation by Extracellular Cues Affects Positioning of the Centrosome and Nucleus and Causes Reduced Cell Migration.

PubMed

Ou, Young; Chan, Gordon; Zuo, Jeremy; Rattner, Jerome B; van der Hoorn, Frans A

2016-07-15

The tight, relative positioning of the nucleus and centrosome in mammalian cells is important for the regulation of cell migration. Under pathophysiological conditions, the purinergic A2b receptor can regulate cell motility, but the underlying mechanism remains unknown. Expression of A2b, normally low, is increased in tissues experiencing adverse physiological conditions, including hypoxia and inflammation. ATP is released from such cells. We investigated whether extracellular cues can regulate centrosome-nucleus positioning and cell migration. We discovered that hypoxia as well as extracellular ATP cause a reversible increase in the distance between the centrosome and nucleus and reduced cell motility. We uncovered the underlying pathway: both treatments act through the A2b receptor and specifically activate the Epac1/RapGef3 pathway. We show that cells lacking A2b do not respond in this manner to hypoxia or ATP but transfection of A2b restores this response, that Epac1 is critically involved, and that Rap1B is important for the relative positioning of the centrosome and nucleus. Our results represent, to our knowledge, the first report demonstrating that pathophysiological conditions can impact the distance between the centrosome and nucleus. Furthermore, we identify the A2b receptor as a central player in this process. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

KENNEDY SPACE CENTER, FLA. -- NASA and United Space Alliance (USA) Space Shuttle program managers attend a briefing, part of activities during a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC. Starting third from left are NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, USA Vice President and Space Shuttle Program Manager Howard DeCastro, NASA Space Shuttle Program Manager William Parsons, and USA Associate Program Manager of Ground Operations Andy Allen.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA and United Space Alliance (USA) Space Shuttle program managers attend a briefing, part of activities during a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC. Starting third from left are NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, USA Vice President and Space Shuttle Program Manager Howard DeCastro, NASA Space Shuttle Program Manager William Parsons, and USA Associate Program Manager of Ground Operations Andy Allen.

Myosin II Motors and F-Actin Dynamics Drive the Coordinated Movement of the Centrosome and Soma during CNS Glial-Guided Neuronal Migration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Solecki, Dr. David; Trivedi, Dr. Niraj; Govek, Eve-Ellen

2009-01-01

Lamination of cortical regions of the vertebrate brain depends on glial-guided neuronal migration. The conserved polarity protein Par6{alpha} localizes to the centrosome and coordinates forward movement of the centrosome and soma in migrating neurons. The cytoskeletal components that produce this unique form of cell polarity and their relationship to polarity signaling cascades are unknown. We show that F-actin and Myosin II motors are enriched in the neuronal leading process and that Myosin II activity is necessary for leading process actin dynamics. Inhibition of Myosin II decreased the speed of centrosome and somal movement, whereas Myosin II activation increased coordinated movement.more » Ectopic expression or silencing of Par6{alpha} inhibited Myosin II motors by decreasing Myosin light-chain phosphorylation. These findings suggest leading-process Myosin II may function to 'pull' the centrosome and soma forward during glial-guided migration by a mechanism involving the conserved polarity protein Par6{alpha}.« less

A functional network involved in the recycling of nucleocytoplasmic pre-60S factors

PubMed Central

Lebreton, Alice; Saveanu, Cosmin; Decourty, Laurence; Rain, Jean-Christophe; Jacquier, Alain; Fromont-Racine, Micheline

2006-01-01

Eukaryotic pre-ribosomes go through cytoplasmic maturation steps before entering translation. The nucleocytoplasmic proteins participating in these late stages of maturation are reimported to the nucleus. In this study, we describe a functional network focused on Rei1/Ybr267w, a strictly cytoplasmic pre-60S factor indirectly involved in nuclear 27S pre-ribosomal RNA processing. In the absence of Rei1, the nuclear import of at least three other pre-60S factors is impaired. The accumulation in the cytoplasm of a small complex formed by the association of Arx1 with a novel factor, Alb1/Yjl122w, inhibits the release of the putative antiassociation factor Tif6 from the premature large ribosomal subunits and its recycling to the nucleus. We propose a model in which Rei1 is a key factor for the coordinated dissociation and recycling of the last pre-60S factors before newly synthesized large ribosomal subunits enter translation. PMID:16651379

CDKL5 localizes at the centrosome and midbody and is required for faithful cell division.

PubMed

Barbiero, Isabella; Valente, Davide; Chandola, Chetan; Magi, Fiorenza; Bergo, Anna; Monteonofrio, Laura; Tramarin, Marco; Fazzari, Maria; Soddu, Silvia; Landsberger, Nicoletta; Rinaldo, Cinzia; Kilstrup-Nielsen, Charlotte

2017-07-24

The cyclin-dependent kinase-like 5 (CDKL5) gene has been associated with rare neurodevelopmental disorders characterized by the early onset of seizures and intellectual disability. The CDKL5 protein is widely expressed in most tissues and cells with both nuclear and cytoplasmic localization. In post-mitotic neurons CDKL5 is mainly involved in dendritic arborization, axon outgrowth, and spine formation while in proliferating cells its function is still largely unknown. Here, we report that CDKL5 localizes at the centrosome and at the midbody in proliferating cells. Acute inactivation of CDKL5 by RNA interference (RNAi) leads to multipolar spindle formation, cytokinesis failure and centrosome accumulation. At the molecular level, we observed that, among the several midbody components we analyzed, midbodies of CDKL5-depleted cells were devoid of HIPK2 and its cytokinesis target, the extrachromosomal histone H2B phosphorylated at S14. Of relevance, expression of the phosphomimetic mutant H2B-S14D, which is capable of overcoming cytokinesis failure in HIPK2-defective cells, was sufficient to rescue spindle multipolarity in CDKL5-depleted cells. Taken together, these results highlight a hitherto unknown role of CDKL5 in regulating faithful cell division by guaranteeing proper HIPK2/H2B functions at the midbody.

Centrosome Amplification: A Potential Marker of Breast Cancer Agressiveness

DTIC Science & Technology

2006-07-01

centrosome amplification. Introduction of DNA damage in the MCF-7 cell line by treatment with hydroxyurea (HU) or daunorubicin (DR) resulted in the...cycles of DNA synthesis and mitotic division in hydroxyurea - arrested Chinese hamster ovary cells. J Cell Biol, 130: 105-115, 1995. 23. D’Assoro, A. B...from cycles of DNA synthesis and mitotic division in hydroxyurea -arrested Chinese hamster ovary cells. J Cell Biol, 1995. 130(1): p. 105-15. 22

Centrosomal Nlp is an oncogenic protein that is gene-amplified in human tumors and causes spontaneous tumorigenesis in transgenic mice

PubMed Central

Shao, Shujuan; Liu, Rong; Wang, Yang; Song, Yongmei; Zuo, Lihui; Xue, Liyan; Lu, Ning; Hou, Ning; Wang, Mingrong; Yang, Xiao; Zhan, Qimin

2010-01-01

Disruption of mitotic events contributes greatly to genomic instability and results in mutator phenotypes. Indeed, abnormalities of mitotic components are closely associated with malignant transformation and tumorigenesis. Here we show that ninein-like protein (Nlp), a recently identified BRCA1-associated centrosomal protein involved in microtubule nucleation and spindle formation, is an oncogenic protein. Nlp was found to be overexpressed in approximately 80% of human breast and lung carcinomas analyzed. In human lung cancers, this deregulated expression was associated with NLP gene amplification. Further analysis revealed that Nlp exhibited strong oncogenic properties; for example, it conferred to NIH3T3 rodent fibroblasts the capacity for anchorage-independent growth in vitro and tumor formation in nude mice. Consistent with these data, transgenic mice overexpressing Nlp displayed spontaneous tumorigenesis in the breast, ovary, and testicle within 60 weeks. In addition, Nlp overexpression induced more rapid onset of radiation-induced lymphoma. Furthermore, mouse embryonic fibroblasts (MEFs) derived from Nlp transgenic mice showed centrosome amplification, suggesting that Nlp overexpression mimics BRCA1 loss. These findings demonstrate that Nlp abnormalities may contribute to genomic instability and tumorigenesis and suggest that Nlp might serve as a potential biomarker for clinical diagnosis and therapeutic target. PMID:20093778

Nesprin 4 is an outer nuclear membrane protein that can induce kinesin-mediated cell polarization.

PubMed

Roux, Kyle J; Crisp, Melissa L; Liu, Qian; Kim, Daein; Kozlov, Serguei; Stewart, Colin L; Burke, Brian

2009-02-17

Nucleocytoplasmic coupling is mediated by outer nuclear membrane (ONM) nesprin proteins and inner nuclear membrane Sun proteins. Interactions spanning the perinuclear space create nesprin-Sun complexes connecting the cytoskeleton to nuclear components. A search for proteins displaying a conserved C-terminal sequence present in nesprins 1-3 identified nesprin 4 (Nesp4), a new member of this family. Nesp4 is a kinesin-1-binding protein that displays Sun-dependent localization to the ONM. Expression of Nesp4 is associated with dramatic changes in cellular organization involving relocation of the centrosome and Golgi apparatus relative to the nucleus. These effects can be accounted for entirely by Nesp4's kinesin-binding function. The implication is that Nesp4 may contribute to microtubule-dependent nuclear positioning.

Cell confinement controls centrosome positioning and lumen initiation during epithelial morphogenesis

PubMed Central

Rodríguez-Fraticelli, Alejo E.; Auzan, Muriel; Alonso, Miguel A.; Bornens, Michel

2012-01-01

Epithelial organ morphogenesis involves sequential acquisition of apicobasal polarity by epithelial cells and development of a functional lumen. In vivo, cells perceive signals from components of the extracellular matrix (ECM), such as laminin and collagens, as well as sense physical conditions, such as matrix stiffness and cell confinement. Alteration of the mechanical properties of the ECM has been shown to promote cell migration and invasion in cancer cells, but the effects on epithelial morphogenesis have not been characterized. We analyzed the effects of cell confinement on lumen morphogenesis using a novel, micropatterned, three-dimensional (3D) Madin-Darby canine kidney cell culture method. We show that cell confinement, by controlling cell spreading, limits peripheral actin contractility and promotes centrosome positioning and lumen initiation after the first cell division. In addition, peripheral actin contractility is mediated by master kinase Par-4/LKB1 via the RhoA–Rho kinase–myosin II pathway, and inhibition of this pathway restores lumen initiation in minimally confined cells. We conclude that cell confinement controls nuclear–centrosomal orientation and lumen initiation during 3D epithelial morphogenesis. PMID:22965908

The presence of centrioles and centrosomes in ovarian mature cystic teratoma cells suggests human parthenotes developed in vitro can differentiate into mature cells without a sperm centriole.

PubMed

Lee, Bo Yon; Shim, Sang Woo; Kim, Young Sun; Kim, Seung Bo

2011-11-18

In most animals, somatic cell centrosomes are inherited from the centriole of the fertilizing spermatozoa. The oocyte centriole degenerates during oogenesis, and completely disappears in metaphase II. Therefore, the embryos generated by in vitro parthenogenesis are supposed to develop without any centrioles. Exceptional acentriolar and/or acentrosomal developments are possible in mice and in some experimental cells; however, in most animals, the full developmental potential of parthenogenetic cells in vitro and the fate of their centrioles/centrosomes are not clearly understood. To predict the future of in vitro human parthenogenesis, we explored the centrioles/centrosomes in ovarian mature cystic teratoma cells by immunofluorescent staining and transmission electron microscopy. We confirmed the presence of centrioles and centrosomes in these well-known parthenogenetic ovarian tumor cells. Our findings clearly demonstrate that, even without a sperm centriole, parthenotes that develop from activated oocytes can produce their own centrioles/centrosomes, and can even develop into the well-differentiated mature tissue. Copyright © 2011 Elsevier Inc. All rights reserved.

Higher Nucleoporin-Importinβ Affinity at the Nuclear Basket Increases Nucleocytoplasmic Import

PubMed Central

Azimi, Mohammad; Mofrad, Mohammad R. K.

2013-01-01

Several in vitro studies have shown the presence of an affinity gradient in nuclear pore complex proteins for the import receptor Importinβ, at least partially contributing to nucleocytoplasmic transport, while others have historically argued against the presence of such a gradient. Nonetheless, the existence of an affinity gradient has remained an uncharacterized contributing factor. To shed light on the affinity gradient theory and better characterize how the existence of such an affinity gradient between the nuclear pore and the import receptor may influence the nucleocytoplasmic traffic, we have developed a general-purpose agent based modeling (ABM) framework that features a new method for relating rate constants to molecular binding and unbinding probabilities, and used our ABM approach to quantify the effects of a wide range of forward and reverse nucleoporin-Importinβ affinity gradients. Our results indicate that transport through the nuclear pore complex is maximized with an effective macroscopic affinity gradient of 2000 µM, 200 µM and 10 µM in the cytoplasmic, central channel and nuclear basket respectively. The transport rate at this gradient is approximately 10% higher than the transport rate for a comparable pore lacking any affinity gradient, which has a peak transport rate when all nucleoporins have an affinity of 200 µM for Importinβ. Furthermore, this optimal ratio of affinity gradients is representative of the ratio of affinities reported for the yeast nuclear pore complex – suggesting that the affinity gradient seen in vitro is highly optimized. PMID:24282617

Identification of a new protein in the centrosome-like "atractophore" of Trichomonas vaginalis.

PubMed

Bricheux, Geneviève; Coffe, Gérard; Brugerolle, Guy

2007-06-01

The human parasite Trichomonas vaginalis has specific structural bodies, atractophores, associated at one end to the kinetosomes and at the other to the spindle during division. A monoclonal antibody specific for a component of this structure was obtained. It recognizes a protein with a predicted molecular mass of 477 kDa. Sequence analysis of this protein shows that P477 belongs to the family of large coiled-coil proteins, sharing a highly versatile protein folding motif adaptable to many biological functions. P477-might act as an anchor to localize cellular activities and components to the golgi centrosomal region. It may represent a new class of structural proteins, since similar proteins were found in many protozoans.

Overexpression of centrosomal protein Nlp confers breast carcinoma resistance to paclitaxel.

PubMed

Zhao, Weihong; Song, Yongmei; Xu, Binghe; Zhan, Qimin

2012-02-01

Nlp (ninein-like protein), an important molecule involved in centrosome maturation and spindle formation, plays an important role in tumorigenesis and its abnormal expression was recently observed in human breast and lung cancers. In this study, the correlation between overexpression of Nlp and paclitaxel chemosensitivity was investigated to explore the mechanisms of resistance to paclitaxel and to understand the effect of Nlp upon apoptosis induced by chemotherapeutic agents. Nlp expression vector was stably transfected into breast cancer MCF-7 cells. With Nlp overexpression, the survival rates, cell cycle distributions and apoptosis were analyzed in transfected MCF-7 cells by MTT test and FCM approach. The immunofluorescent assay was employed to detect the changes of microtubule after paclitaxel treatment. Immunoblotting analysis was used to examine expression of centrosomal proteins and apoptosis associated proteins. Subsequently, Nlp expression was retrospectively examined with 55 breast cancer samples derived from paclitaxel treated patients. Interestingly, the survival rates of MCF-7 cells with Nlp overexpressing were higher than that of control after paclitaxel treatment. Nlp overexpression promoted G2-M arrest and attenuated apoptosis induced by paclitaxel, which was coupled with elevated Bcl-2 protein. Nlp expression significantly lessened the microtubule polymerization and bundling elicited by paclitaxel attributing to alteration on the structure or dynamics of β-tubulin but not on its expression. The breast cancer patients with high expression of Nlp were likely resistant to the treatment of paclitaxel, as the response rate in Nlp negative patients was 62.5%, whereas was 58.3 and 15.8% in Nlp (+) and Nlp (++) patients respectively (p = 0.015). Nlp expression was positive correlated with those of Plk1 and PCNA. These findings provide insights into more rational chemotherapeutic regimens in clinical practice, and more effective approaches might be

Concerted effort of centrosomal and Golgi-derived microtubules is required for proper Golgi complex assembly but not for maintenance

PubMed Central

Vinogradova, Tatiana; Paul, Raja; Grimaldi, Ashley D.; Loncarek, Jadranka; Miller, Paul M.; Yampolsky, Dmitry; Magidson, Valentin; Khodjakov, Alexey; Mogilner, Alex; Kaverina, Irina

2012-01-01

Assembly of an integral Golgi complex is driven by microtubule (MT)-dependent transport. Conversely, the Golgi itself functions as an unconventional MT-organizing center (MTOC). This raises the question of whether Golgi assembly requires centrosomal MTs or can be self-organized, relying on its own MTOC activity. The computational model presented here predicts that each MT population is capable of gathering Golgi stacks but not of establishing Golgi complex integrity or polarity. In contrast, the concerted effort of two MT populations would assemble an integral, polarized Golgi complex. Indeed, while laser ablation of the centrosome did not alter already-formed Golgi complexes, acentrosomal cells fail to reassemble an integral complex upon nocodazole washout. Moreover, polarity of post-Golgi trafficking was compromised under these conditions, leading to strong deficiency in polarized cell migration. Our data indicate that centrosomal MTs complement Golgi self-organization for proper Golgi assembly and motile-cell polarization. PMID:22262454

Exploring nucleo-cytoplasmic large DNA viruses in Tara Oceans microbial metagenomes

PubMed Central

Hingamp, Pascal; Grimsley, Nigel; Acinas, Silvia G; Clerissi, Camille; Subirana, Lucie; Poulain, Julie; Ferrera, Isabel; Sarmento, Hugo; Villar, Emilie; Lima-Mendez, Gipsi; Faust, Karoline; Sunagawa, Shinichi; Claverie, Jean-Michel; Moreau, Hervé; Desdevises, Yves; Bork, Peer; Raes, Jeroen; de Vargas, Colomban; Karsenti, Eric; Kandels-Lewis, Stefanie; Jaillon, Olivier; Not, Fabrice; Pesant, Stéphane; Wincker, Patrick; Ogata, Hiroyuki

2013-01-01

Nucleo-cytoplasmic large DNA viruses (NCLDVs) constitute a group of eukaryotic viruses that can have crucial ecological roles in the sea by accelerating the turnover of their unicellular hosts or by causing diseases in animals. To better characterize the diversity, abundance and biogeography of marine NCLDVs, we analyzed 17 metagenomes derived from microbial samples (0.2–1.6 μm size range) collected during the Tara Oceans Expedition. The sample set includes ecosystems under-represented in previous studies, such as the Arabian Sea oxygen minimum zone (OMZ) and Indian Ocean lagoons. By combining computationally derived relative abundance and direct prokaryote cell counts, the abundance of NCLDVs was found to be in the order of 104–105 genomes ml−1 for the samples from the photic zone and 102–103 genomes ml−1 for the OMZ. The Megaviridae and Phycodnaviridae dominated the NCLDV populations in the metagenomes, although most of the reads classified in these families showed large divergence from known viral genomes. Our taxon co-occurrence analysis revealed a potential association between viruses of the Megaviridae family and eukaryotes related to oomycetes. In support of this predicted association, we identified six cases of lateral gene transfer between Megaviridae and oomycetes. Our results suggest that marine NCLDVs probably outnumber eukaryotic organisms in the photic layer (per given water mass) and that metagenomic sequence analyses promise to shed new light on the biodiversity of marine viruses and their interactions with potential hosts. PMID:23575371

Exploring nucleo-cytoplasmic large DNA viruses in Tara Oceans microbial metagenomes.

PubMed

Hingamp, Pascal; Grimsley, Nigel; Acinas, Silvia G; Clerissi, Camille; Subirana, Lucie; Poulain, Julie; Ferrera, Isabel; Sarmento, Hugo; Villar, Emilie; Lima-Mendez, Gipsi; Faust, Karoline; Sunagawa, Shinichi; Claverie, Jean-Michel; Moreau, Hervé; Desdevises, Yves; Bork, Peer; Raes, Jeroen; de Vargas, Colomban; Karsenti, Eric; Kandels-Lewis, Stefanie; Jaillon, Olivier; Not, Fabrice; Pesant, Stéphane; Wincker, Patrick; Ogata, Hiroyuki

2013-09-01

Nucleo-cytoplasmic large DNA viruses (NCLDVs) constitute a group of eukaryotic viruses that can have crucial ecological roles in the sea by accelerating the turnover of their unicellular hosts or by causing diseases in animals. To better characterize the diversity, abundance and biogeography of marine NCLDVs, we analyzed 17 metagenomes derived from microbial samples (0.2-1.6 μm size range) collected during the Tara Oceans Expedition. The sample set includes ecosystems under-represented in previous studies, such as the Arabian Sea oxygen minimum zone (OMZ) and Indian Ocean lagoons. By combining computationally derived relative abundance and direct prokaryote cell counts, the abundance of NCLDVs was found to be in the order of 10(4)-10(5) genomes ml(-1) for the samples from the photic zone and 10(2)-10(3) genomes ml(-1) for the OMZ. The Megaviridae and Phycodnaviridae dominated the NCLDV populations in the metagenomes, although most of the reads classified in these families showed large divergence from known viral genomes. Our taxon co-occurrence analysis revealed a potential association between viruses of the Megaviridae family and eukaryotes related to oomycetes. In support of this predicted association, we identified six cases of lateral gene transfer between Megaviridae and oomycetes. Our results suggest that marine NCLDVs probably outnumber eukaryotic organisms in the photic layer (per given water mass) and that metagenomic sequence analyses promise to shed new light on the biodiversity of marine viruses and their interactions with potential hosts.

Androgen and taxol cause cell type-specific alterations of centrosome and DNA organization in androgen-responsive LNCaP and androgen-independent DU145 prostate cancer cells

NASA Technical Reports Server (NTRS)

Schatten, H.; Ripple, M.; Balczon, R.; Weindruch, R.; Chakrabarti, A.; Taylor, M.; Hueser, C. N.

2000-01-01

We investigated the effects of androgen and taxol on the androgen-responsive LNCaP and androgen-independent DU145 prostate cancer cell lines. Cells were treated for 48 and 72 h with 0.05-1 nM of the synthetic androgen R1881 and with 100 nM taxol. Treatment of LNCaP cells with 0.05 nM R1881 led to increased cell proliferation, whereas treatment with 1 nM R1881 resulted in inhibited cell division, DNA cycle arrest, and altered centrosome organization. After treatment with 1 nM R1881, chromatin became clustered, nuclear envelopes convoluted, and mitochondria accumulated around the nucleus. Immunofluorescence microscopy with antibodies to centrosomes showed altered centrosome structure. Although centrosomes were closely associated with the nucleus in untreated cells, they dispersed into the cytoplasm after treatment with 1 nM R1881. Microtubules were only faintly detected in 1 nM R1881-treated LNCaP cells. The effects of taxol included microtubule bundling and altered mitochondria morphology, but not DNA organization. As expected, the androgen-independent prostate cancer cell line DU145 was not affected by R1881. Treatment with taxol resulted in bundling of microtubules in both cell lines. Additional taxol effects were seen in DU145 cells with micronucleation of DNA, an indication of apoptosis. Simultaneous treatment with R1881 and taxol had no additional effects on LNCaP or DU145 cells. These results suggest that LNCaP and DU145 prostate cancer cells show differences not only in androgen responsiveness but in sensitivity to taxol as well. Copyright 2000 Wiley-Liss, Inc.

Nesprin 4 is an outer nuclear membrane protein that can induce kinesin-mediated cell polarization

PubMed Central

Roux, Kyle J.; Crisp, Melissa L.; Liu, Qian; Kim, Daein; Kozlov, Serguei; Stewart, Colin L.; Burke, Brian

2009-01-01

Nucleocytoplasmic coupling is mediated by outer nuclear membrane (ONM) nesprin proteins and inner nuclear membrane Sun proteins. Interactions spanning the perinuclear space create nesprin–Sun complexes connecting the cytoskeleton to nuclear components. A search for proteins displaying a conserved C-terminal sequence present in nesprins 1–3 identified nesprin 4 (Nesp4), a new member of this family. Nesp4 is a kinesin-1-binding protein that displays Sun-dependent localization to the ONM. Expression of Nesp4 is associated with dramatic changes in cellular organization involving relocation of the centrosome and Golgi apparatus relative to the nucleus. These effects can be accounted for entirely by Nesp4's kinesin-binding function. The implication is that Nesp4 may contribute to microtubule-dependent nuclear positioning. PMID:19164528

An Amino-Terminal Polo Kinase Interaction Motif Acts in the Regulation of Centrosome Formation and Reveals a Novel Function for centrosomin (cnn) in Drosophila

PubMed Central

Eisman, Robert C.; Phelps, Melissa A. S.; Kaufman, Thomas

2015-01-01

The formation of the pericentriolar matrix (PCM) and a fully functional centrosome in syncytial Drosophila melanogaster embryos requires the rapid transport of Cnn during initiation of the centrosome replication cycle. We show a Cnn and Polo kinase interaction is apparently required during embryogenesis and involves the exon 1A-initiating coding exon, suggesting a subset of Cnn splice variants is regulated by Polo kinase. During PCM formation exon 1A Cnn-Long Form proteins likely bind Polo kinase before phosphorylation by Polo for Cnn transport to the centrosome. Loss of either of these interactions in a portion of the total Cnn protein pool is sufficient to remove native Cnn from the pool, thereby altering the normal localization dynamics of Cnn to the PCM. Additionally, Cnn-Short Form proteins are required for polar body formation, a process known to require Polo kinase after the completion of meiosis. Exon 1A Cnn-LF and Cnn-SF proteins, in conjunction with Polo kinase, are required at the completion of meiosis and for the formation of functional centrosomes during early embryogenesis. PMID:26447129

Space Shuttle

NASA Technical Reports Server (NTRS)

1975-01-01

A general description of the space shuttle program is presented, with emphasis on its application to the use of space for commercial, scientific, and defense needs. The following aspects of the program are discussed: description of the flight system (orbiter, external tank, solid rocket boosters) and mission profile, direct benefits related to life on earth (both present and expected), description of the space shuttle vehicle and its associated supporting systems, economic impacts (including indirect benefits such as lower inflation rates), listing of participating organizations.

Centrosome Hypertrophy Induced by p53 Mutations Leads to Tumor Aneuploidy

DTIC Science & Technology

1999-06-01

6). A tripolar mitosis is shown in Figure 6A. Tracings of microtubules, spindle poles, and condensed chromosomes from six non-adjacent serial sections...Centrin Immunofluorescence. A. This section through a symmetrical tripolar mitotic cell shows part of the metaphase plate and portions of the tripolar ...A tripolar metaphase cell immunolabeled with Ki-67 is shown in this tumor section. E. In normal breast epithelium, the centrosomes appear as distinct

The conserved Wdr8-hMsd1/SSX2IP complex localises to the centrosome and ensures proper spindle length and orientation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hori, Akiko; Morand, Agathe; Ikebe, Chiho

2015-12-04

The centrosome plays a pivotal role in a wide range of cellular processes and its dysfunction is causally linked to many human diseases including cancer and developmental and neurological disorders. This organelle contains more than one hundred components, and yet many of them remain uncharacterised. Here we identified a novel centrosome protein Wdr8, based upon the structural conservation of the fission yeast counterpart. We showed that Wdr8 constitutively localises to the centrosome and super resolution microscopy uncovered that this protein is enriched at the proximal end of the mother centriole. Furthermore, we identified hMsd1/SSX2IP, a conserved spindle anchoring protein, asmore » one of Wdr8 interactors by mass spectrometry. Wdr8 formed a complex and partially colocalised with hMsd1/SSX2IP. Intriguingly, knockdown of Wdr8 or hMsd1/SSX2IP displayed very similar mitotic defects, in which spindle microtubules became shortened and misoriented. Indeed, Wdr8 depletion resulted in the reduced recruitment of hMsd1/SSX2IP to the mitotic centrosome, though the converse is not true. Together, we propose that the conserved Wdr8-hMsd1/SSX2IP complex plays a critical role in controlling proper spindle length and orientation. - Highlights: • Human Wdr8 is a centrosomal protein enriched in the proximal end of the centriole. • Wdr8 and hMsd1/SSX2IP form a complex conserved in fungi. • Depletion of Wdr8 results in shorter, tilted spindle microtubules. • Depletion phenotypes of Wdr8 are very similar to those of hMsd1/SSX2IP knockdown.« less

The conserved Wdr8-hMsd1/SSX2IP complex localises to the centrosome and ensures proper spindle length and orientation

PubMed Central

Hori, Akiko; Morand, Agathe; Ikebe, Chiho; Frith, David; Snijders, Ambrosius P.; Toda, Takashi

2015-01-01

The centrosome plays a pivotal role in a wide range of cellular processes and its dysfunction is causally linked to many human diseases including cancer and developmental and neurological disorders. This organelle contains more than one hundred components, and yet many of them remain uncharacterised. Here we identified a novel centrosome protein Wdr8, based upon the structural conservation of the fission yeast counterpart. We showed that Wdr8 constitutively localises to the centrosome and super resolution microscopy uncovered that this protein is enriched at the proximal end of the mother centriole. Furthermore, we identified hMsd1/SSX2IP, a conserved spindle anchoring protein, as one of Wdr8 interactors by mass spectrometry. Wdr8 formed a complex and partially colocalised with hMsd1/SSX2IP. Intriguingly, knockdown of Wdr8 or hMsd1/SSX2IP displayed very similar mitotic defects, in which spindle microtubules became shortened and misoriented. Indeed, Wdr8 depletion resulted in the reduced recruitment of hMsd1/SSX2IP to the mitotic centrosome, though the converse is not true. Together, we propose that the conserved Wdr8-hMsd1/SSX2IP complex plays a critical role in controlling proper spindle length and orientation. PMID:26545777

Shuttle Entry Imaging Using Infrared Thermography

NASA Technical Reports Server (NTRS)

Horvath, Thomas; Berry, Scott; Alter, Stephen; Blanchard, Robert; Schwartz, Richard; Ross, Martin; Tack, Steve

2007-01-01

During the Columbia Accident Investigation, imaging teams supporting debris shedding analysis were hampered by poor entry image quality and the general lack of information on optical signatures associated with a nominal Shuttle entry. After the accident, recommendations were made to NASA management to develop and maintain a state-of-the-art imagery database for Shuttle engineering performance assessments and to improve entry imaging capability to support anomaly and contingency analysis during a mission. As a result, the Space Shuttle Program sponsored an observation campaign to qualitatively characterize a nominal Shuttle entry over the widest possible Mach number range. The initial objectives focused on an assessment of capability to identify/resolve debris liberated from the Shuttle during entry, characterization of potential anomalous events associated with RCS jet firings and unusual phenomenon associated with the plasma trail. The aeroheating technical community viewed the Space Shuttle Program sponsored activity as an opportunity to influence the observation objectives and incrementally demonstrate key elements of a quantitative spatially resolved temperature measurement capability over a series of flights. One long-term desire of the Shuttle engineering community is to calibrate boundary layer transition prediction methodologies that are presently part of the Shuttle damage assessment process using flight data provided by a controlled Shuttle flight experiment. Quantitative global imaging may offer a complementary method of data collection to more traditional methods such as surface thermocouples. This paper reviews the process used by the engineering community to influence data collection methods and analysis of global infrared images of the Shuttle obtained during hypersonic entry. Emphasis is placed upon airborne imaging assets sponsored by the Shuttle program during Return to Flight. Visual and IR entry imagery were obtained with available airborne

A Phthalimide Derivative That Inhibits Centrosomal Clustering Is Effective on Multiple Myeloma

PubMed Central

Shiheido, Hirokazu; Terada, Fukiko; Tabata, Noriko; Hayakawa, Ichigo; Matsumura, Nobutaka; Takashima, Hideaki; Ogawa, Yoko; Du, Wenlin; Yamada, Taketo; Shoji, Mitsuru; Sugai, Takeshi; Doi, Nobuhide; Iijima, Shiro; Hattori, Yutaka; Yanagawa, Hiroshi

2012-01-01

Despite the introduction of newly developed drugs such as lenalidomide and bortezomib, patients with multiple myeloma are still difficult to treat and have a poor prognosis. In order to find novel drugs that are effective for multiple myeloma, we tested the antitumor activity of 29 phthalimide derivatives against several multiple myeloma cell lines. Among these derivatives, 2-(2,6-diisopropylphenyl)-5-amino-1H-isoindole-1,3- dione (TC11) was found to be a potent inhibitor of tumor cell proliferation and an inducer of apoptosis via activation of caspase-3, 8 and 9. This compound also showed in vivo activity against multiple myeloma cell line KMS34 tumor xenografts in ICR/SCID mice. By means of mRNA display selection on a microfluidic chip, the target protein of TC11 was identified as nucleophosmin 1 (NPM). Binding of TC11 and NPM monomer was confirmed by surface plasmon resonance. Immunofluorescence and NPM knockdown studies in HeLa cells suggested that TC11 inhibits centrosomal clustering by inhibiting the centrosomal-regulatory function of NPM, thereby inducing multipolar mitotic cells, which undergo apoptosis. NPM may become a novel target for development of antitumor drugs active against multiple myeloma. PMID:22761710

An Amino-Terminal Polo Kinase Interaction Motif Acts in the Regulation of Centrosome Formation and Reveals a Novel Function for centrosomin (cnn) in Drosophila.

PubMed

Eisman, Robert C; Phelps, Melissa A S; Kaufman, Thomas

2015-10-01

The formation of the pericentriolar matrix (PCM) and a fully functional centrosome in syncytial Drosophila melanogaster embryos requires the rapid transport of Cnn during initiation of the centrosome replication cycle. We show a Cnn and Polo kinase interaction is apparently required during embryogenesis and involves the exon 1A-initiating coding exon, suggesting a subset of Cnn splice variants is regulated by Polo kinase. During PCM formation exon 1A Cnn-Long Form proteins likely bind Polo kinase before phosphorylation by Polo for Cnn transport to the centrosome. Loss of either of these interactions in a portion of the total Cnn protein pool is sufficient to remove native Cnn from the pool, thereby altering the normal localization dynamics of Cnn to the PCM. Additionally, Cnn-Short Form proteins are required for polar body formation, a process known to require Polo kinase after the completion of meiosis. Exon 1A Cnn-LF and Cnn-SF proteins, in conjunction with Polo kinase, are required at the completion of meiosis and for the formation of functional centrosomes during early embryogenesis. Copyright © 2015 by the Genetics Society of America.

Nucleocytoplasmic Transport: A Paradigm for Molecular Logistics in Artificial Systems.

PubMed

Vujica, Suncica; Zelmer, Christina; Panatala, Radhakrishnan; Lim, Roderick Y H

2016-01-01

Artificial organelles, molecular factories and nanoreactors are membrane-bound systems envisaged to exhibit cell-like functionality. These constitute liposomes, polymersomes or hybrid lipo-polymersomes that display different membrane-spanning channels and/or enclose molecular modules. To achieve more complex functionality, an artificial organelle should ideally sustain a continuous influx of essential macromolecular modules (i.e. cargoes) and metabolites against an outflow of reaction products. This would benefit from the incorporation of selective nanopores as well as specific trafficking factors that facilitate cargo selectivity, translocation efficiency, and directionality. Towards this goal, we describe how proteinaceous cargoes are transported between the nucleus and cytoplasm by nuclear pore complexes and the biological trafficking machinery in living cells (i.e. nucleocytoplasmic transport). On this basis, we discuss how biomimetic control may be implemented to selectively import, compartmentalize and accumulate diverse macromolecular modules against concentration gradients in artificial organelles.

The role of mammalian Staufen on mRNA traffic: a view from its nucleocytoplasmic shuttling function.

PubMed

Miki, Takashi; Takano, Keizo; Yoneda, Yoshihiro

2005-01-01

The localization of mRNA in neuronal dendrites plays a role in both locally and temporally regulated protein synthesis, which is required for certain forms of synaptic plasticity. RNA granules constitute a dendritic mRNA transport machinery in neurons, which move along microtubules. RNA granules contain densely packed clusters of ribosomes, but lack some factors that are required for translation, suggesting that they are translationally incompetent. Recently some of the components of RNA granules have been identified, and their functions are in the process of being examined, in attempts to better understand the properties of RNA granules. Mammalian Staufen, a double-stranded RNA binding protein, is a component of RNA granules. Staufen is localized in the somatodendritic domain of neurons, and plays an important role in dendritic mRNA targeting. Recently, one of the mammalian homologs of Staufen, Staufen2 (Stau2), was shown to shuttle between the nucleus and the cytoplasm. This finding suggests the possibility that Stau2 binds RNA in the nucleus and that this ribonucleoprotein particle is transported from the nucleus to RNA granules in the cytoplasm. A closer study of this process might provide a clue to the mechanism by which RNA granules are formed.

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, United Space Alliance (USA) Director of Orbiter Operations Patty Stratton, and NASA Space Shuttle Program Manager William Parsons view the underside of Shuttle Discovery in Orbiter Processing Facility Bay 3. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, United Space Alliance (USA) Director of Orbiter Operations Patty Stratton, and NASA Space Shuttle Program Manager William Parsons view the underside of Shuttle Discovery in Orbiter Processing Facility Bay 3. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Space Shuttle Usage of z/OS

NASA Technical Reports Server (NTRS)

Green, Jan

2009-01-01

This viewgraph presentation gives a detailed description of the avionics associated with the Space Shuttle's data processing system and its usage of z/OS. The contents include: 1) Mission, Products, and Customers; 2) Facility Overview; 3) Shuttle Data Processing System; 4) Languages and Compilers; 5) Application Tools; 6) Shuttle Flight Software Simulator; 7) Software Development and Build Tools; and 8) Fun Facts and Acronyms.

STS-121 Space Shuttle Processing Update

NASA Image and Video Library

2006-04-27

NASA Administrator Michael Griffin, left, and Associate Administrator for Space Operations William Gerstenmaier, right, look on as Space Shuttle Program Manager Wayne Hale talks from NASA's Marshall Space Flight Center about the space shuttle's ice frost ramps during a media briefing about the space shuttle program and processing for the STS-121 mission, Friday, April 28, 2006, at NASA Headquarters in Washington. Photo Credit (NASA/Bill Ingalls)

MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore.

PubMed

Ren, Jian; Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

2010-01-01

During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0).

The role of centrosomal Nlp in the control of mitotic progression and tumourigenesis.

PubMed

Li, J; Zhan, Q

2011-05-10

The human centrosomal ninein-like protein (Nlp) is a new member of the γ-tubulin complexes binding proteins (GTBPs) that is essential for proper execution of various mitotic events. The primary function of Nlp is to promote microtubule nucleation that contributes to centrosome maturation, spindle formation and chromosome segregation. Its subcellular localisation and protein stability are regulated by several crucial mitotic kinases, such as Plk1, Nek2, Cdc2 and Aurora B. Several lines of evidence have linked Nlp to human cancer. Deregulation of Nlp in cell models results in aberrant spindle, chromosomal missegregation and multinulei, and induces chromosomal instability and renders cells tumourigenic. Overexpression of Nlp induces anchorage-independent growth and immortalised primary cell transformation. In addition, we first demonstrate that the expression of Nlp is elevated primarily due to NLP gene amplification in human breast cancer and lung carcinoma. Consistently, transgenic mice overexpressing Nlp display spontaneous tumours in breast, ovary and testicle, and show rapid onset of radiation-induced lymphoma, indicating that Nlp is involved in tumourigenesis. This review summarises our current knowledge of physiological roles of Nlp, with an emphasis on its potentials in tumourigenesis.

The role of centrosomal Nlp in the control of mitotic progression and tumourigenesis

PubMed Central

Li, J; Zhan, Q

2011-01-01

The human centrosomal ninein-like protein (Nlp) is a new member of the γ-tubulin complexes binding proteins (GTBPs) that is essential for proper execution of various mitotic events. The primary function of Nlp is to promote microtubule nucleation that contributes to centrosome maturation, spindle formation and chromosome segregation. Its subcellular localisation and protein stability are regulated by several crucial mitotic kinases, such as Plk1, Nek2, Cdc2 and Aurora B. Several lines of evidence have linked Nlp to human cancer. Deregulation of Nlp in cell models results in aberrant spindle, chromosomal missegregation and multinulei, and induces chromosomal instability and renders cells tumourigenic. Overexpression of Nlp induces anchorage-independent growth and immortalised primary cell transformation. In addition, we first demonstrate that the expression of Nlp is elevated primarily due to NLP gene amplification in human breast cancer and lung carcinoma. Consistently, transgenic mice overexpressing Nlp display spontaneous tumours in breast, ovary and testicle, and show rapid onset of radiation-induced lymphoma, indicating that Nlp is involved in tumourigenesis. This review summarises our current knowledge of physiological roles of Nlp, with an emphasis on its potentials in tumourigenesis. PMID:21505454

How to be good at being bad: centrosome amplification and mitotic propensity drive intratumoral heterogeneity

PubMed Central

Rida, Padmashree C. G.; Cantuaria, Guilherme; Reid, Michelle D.; Kucuk, Omer

2016-01-01

Cancer is truly an iconic disease—a tour de force whose multiple formidable strengths can be attributed to the bewildering heterogeneity that a tumor can manifest both spatially and temporally. A Darwinian evolutionary process is believed to undergird, at least in part, the generation of this heterogeneity that contributes to poor clinical outcomes. Risk assessment in clinical oncology is currently based on a small number of clinicopathologic factors (like stage, histological grade, receptor status, and serum tumor markers) and offers limited accuracy in predicting disease course as evidenced by the prognostic heterogeneity that persists in risk segments produced by present-day models. We posit that this insufficiency stems from the exclusion of key risk contributors from such models, especially the omission of certain factors implicated in generating intratumoral heterogeneity. The extent of centrosome amplification and the mitotic propensity inherent in a tumor are two such vital factors whose contributions to poor prognosis are presently overlooked in risk prognostication. Supernumerary centrosomes occur widely in tumors and are potent drivers of chromosomal instability that fosters intratumoral heterogeneity. The mitotic propensity of a proliferating population of tumor cells reflects the cell cycling kinetics of that population. Since frequent passage through improperly regulated mitotic divisions accelerates production of diverse genotypes, the mitotic propensity inherent in a tumor serves as a powerful beacon of risk. In this review, we highlight how centrosome amplification and error-prone mitoses contribute to poor clinical outcomes and urge the need to develop these cancer-specific traits as much-needed clinically-facile prognostic biomarkers with immense potential value for individualized cancer treatment in the clinic. PMID:26358854

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (top) discusses the inner workings of Shuttle Atlantis in Orbiter Processing Facility Bay 1 with a United Space Alliance (USA) technician (bottom). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (top) discusses the inner workings of Shuttle Atlantis in Orbiter Processing Facility Bay 1 with a United Space Alliance (USA) technician (bottom). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right) discusses a speed brake on Shuttle Discovery in Orbiter Processing Facility Bay 3 with a United Space Alliance (USA) technician (left). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right) discusses a speed brake on Shuttle Discovery in Orbiter Processing Facility Bay 3 with a United Space Alliance (USA) technician (left). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Purification, isolation, crystallization, and preliminary X-ray diffraction study of the BTB domain of the centrosomal protein 190 from Drosophila melanogaster

NASA Astrophysics Data System (ADS)

Boyko, K. M.; Nikolaeva, A. Yu.; Kachalova, G. S.; Bonchuk, A. N.; Popov, V. O.

2017-11-01

The spatial organization of the genome is controlled by a special class of architectural proteins, including proteins containing BTB domains that are able to dimerize or multimerize. The centrosomal protein 190 is one of such architectural proteins. The purification, crystallization, and preliminary X-ray diffraction study of the BTB domain of the centrosomal protein 190 are reported. The crystallization conditions were found by the vapor-diffusion technique. The crystals diffracted to 1.5 Å resolution and belonged to sp. gr. P3221. The structure was solved by the molecular replacement method. The structure refinement is currently underway.

Review of Issues Associated with Safe Operation and Management of the Space Shuttle Program

NASA Technical Reports Server (NTRS)

Johnstone, Paul M.; Blomberg, Richard D.; Gleghorn, George J.; Krone, Norris J.; Voltz, Richard A.; Dunn, Robert F.; Donlan, Charles J.; Kauderer, Bernard M.; Brill, Yvonne C.; Englar, Kenneth G.;

Space Shuttle Glider. Educational Brief.

ERIC Educational Resources Information Center

National Aeronautics and Space Administration, Washington, DC.

Space Shuttle Glider is a scale model of the U.S. Space Shuttle orbiter. The airplane-like orbiter usually remains in Earth orbit for up to two weeks at a time. It normally carries a six- to seven-person crew which includes the mission commander, pilot, and several mission and/or payload specialists who have specialized training associated with…

The conversion of centrioles to centrosomes: essential coupling of duplication with segregation.

PubMed

Wang, Won-Jing; Soni, Rajesh Kumar; Uryu, Kunihiro; Tsou, Meng-Fu Bryan

2011-05-16

Centrioles are self-reproducing organelles that form the core structure of centrosomes or microtubule-organizing centers (MTOCs). However, whether duplication and MTOC organization reflect innate activities of centrioles or activities acquired conditionally is unclear. In this paper, we show that newly formed full-length centrioles had no inherent capacity to duplicate or to organize pericentriolar material (PCM) but acquired both after mitosis through a Plk1-dependent modification that occurred in early mitosis. Modified centrioles initiated PCM recruitment in G1 and segregated equally in mitosis through association with spindle poles. Conversely, unmodified centrioles segregated randomly unless passively tethered to modified centrioles. Strikingly, duplication occurred only in centrioles that were both modified and disengaged, whereas unmodified centrioles, engaged or not, were "infertile," indicating that engagement specifically blocks modified centrioles from reduplication. These two requirements, centriole modification and disengagement, fully exclude unlimited duplication in one cell cycle. We thus uncovered a Plk1-dependent mechanism whereby duplication and segregation are coupled to maintain centriole homeostasis.

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) technicians demonstrate the construction of a thermal blanket used in the Shuttle's thermal protection system for USA Vice President and Space Shuttle Program Manager Howard DeCastro (second from left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) technicians demonstrate the construction of a thermal blanket used in the Shuttle's thermal protection system for USA Vice President and Space Shuttle Program Manager Howard DeCastro (second from left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- From left, a United Space Alliance (USA) technician discusses aspects of Shuttle processing performed in the Solid Rocket Booster (SRB) Assembly and Refurbishment Facility (ARF) with USA Vice President and Space Shuttle Program Manager Howard DeCastro and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, a United Space Alliance (USA) technician discusses aspects of Shuttle processing performed in the Solid Rocket Booster (SRB) Assembly and Refurbishment Facility (ARF) with USA Vice President and Space Shuttle Program Manager Howard DeCastro and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (third from left) watch as a USA technician (right) creates a tile for use in the Shuttle's Thermal Protection System (TPS). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (third from left) watch as a USA technician (right) creates a tile for use in the Shuttle's Thermal Protection System (TPS). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Axin localizes to mitotic spindles and centrosomes in mitotic cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Shi-Mun; Choi, Eun-Jin; Song, Ki-Joon

2009-04-01

Wnt signaling plays critical roles in cell proliferation and carcinogenesis. In addition, numerous recent studies have shown that various Wnt signaling components are involved in mitosis and chromosomal instability. However, the role of Axin, a negative regulator of Wnt signaling, in mitosis has remained unclear. Using monoclonal antibodies against Axin, we found that Axin localizes to the centrosome and along mitotic spindles. This localization was suppressed by siRNA specific for Aurora A kinase and by Aurora kinase inhibitor. Interestingly, Axin over-expression altered the subcellular distribution of Plk1 and of phosphorylated glycogen synthase kinase (GSK3{beta}) without producing any notable changes inmore » cellular phenotype. In the presence of Aurora kinase inhibitor, Axin over-expression induced the formation of cleavage furrow-like structures and of prominent astral microtubules lacking midbody formation in a subset of cells. Our results suggest that Axin modulates distribution of Axin-associated proteins such as Plk1 and GSK3{beta} in an expression level-dependent manner and these interactions affect the mitotic process, including cytokinesis under certain conditions, such as in the presence of Aurora kinase inhibitor.« less

MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore

PubMed Central

Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

2010-01-01

During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0). PMID:19783819

GAK, a regulator of clathrin-mediated membrane traffic, also controls centrosome integrity and chromosome congression.

PubMed

Shimizu, Hiroyuki; Nagamori, Ippei; Yabuta, Norikazu; Nojima, Hiroshi

2009-09-01

Cyclin G-associated kinase (GAK) is an association partner of clathrin heavy chain (CHC) and is essential for clathrin-mediated membrane trafficking. Here, we report two novel functions of GAK: maintenance of proper centrosome maturation and of mitotic chromosome congression. Indeed, GAK knockdown by siRNA caused cell-cycle arrest at metaphase, which indicates that GAK is required for proper mitotic progression. We found that this impaired mitotic progression was due to activation of the spindle-assembly checkpoint, which senses protruded, misaligned or abnormally condensed chromosomes in GAK-siRNA-treated cells. GAK knockdown also caused multi-aster formation, which was due to abnormal fragmentation of pericentriolar material, but not of the centrioles. Moreover, GAK and CHC cooperated in the same pathway and interacted in mitosis to regulate the formation of a functional spindle. Taken together, we conclude that GAK and clathrin function cooperatively not only in endocytosis, but also in mitotic progression.

Space Shuttle Projects

NASA Image and Video Library

1991-04-05

Aboard the Space Shuttle Atlantis, the STS-37 mission launched April 5, 1991 from launch pad 39B at the Kennedy Space Center in Florida, and landed back on Earth April 11, 1991. The 39th shuttle mission included crew members: Steven R. Nagel, commander; Kenneth D. Cameron, pilot; Jerry L,. Ross, mission specialist 1; Jay Apt, mission specialist 2; and Linda M. Godwin, mission specialist 3. The primary payload for the mission was the Gamma Ray Observatory (GRO). The GRO included the Burst and Transient Experiment (BATSE); the Imaging Compton Telescope (COMPTEL); the Energetic Gamma Ray Experiment Telescope (EGRET); and the Oriented Scintillation Spectrometer Experiment (OSSEE). Secondary payloads included Crew and Equipment Translation Aids (CETA); the Ascent Particle Monitor (APM); the Shuttle Amateur Radio Experiment II (SAREXII), the Protein Crystal Growth (PCG); the Bioserve Instrumentation Technology Associates Materials Dispersion Apparatus (BIMDA); Radiation Monitoring Equipment III (RMEIII); and Air Force Maui Optical Site (AMOS).

KENNEDY SPACE CENTER, FLA. -- A United Space Alliance (USA) technician (center) discusses aspects of Shuttle processing performed in the Solid Rocket Booster (SRB) Assembly and Refurbishment Facility (ARF) with NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- A United Space Alliance (USA) technician (center) discusses aspects of Shuttle processing performed in the Solid Rocket Booster (SRB) Assembly and Refurbishment Facility (ARF) with NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Space-Shuttle Emulator Software

NASA Technical Reports Server (NTRS)

Arnold, Scott; Askew, Bill; Barry, Matthew R.; Leigh, Agnes; Mermelstein, Scott; Owens, James; Payne, Dan; Pemble, Jim; Sollinger, John; Thompson, Hiram;

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Program Manager of the Space Shuttle Program Michael Wetmore, United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, and a USA technician examine cold plates in Orbiter Processing Facility Bay 2. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Program Manager of the Space Shuttle Program Michael Wetmore, United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik, and a USA technician examine cold plates in Orbiter Processing Facility Bay 2. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- From left, a United Space Alliance (USA) technician briefs NASA Deputy Program Manager of the Space Shuttle Program Michael Wetmore, USA Vice President and Space Shuttle Program Manager Howard DeCastro, and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik on the use of cold plates in Orbiter Processing Facility Bay 2. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, a United Space Alliance (USA) technician briefs NASA Deputy Program Manager of the Space Shuttle Program Michael Wetmore, USA Vice President and Space Shuttle Program Manager Howard DeCastro, and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik on the use of cold plates in Orbiter Processing Facility Bay 2. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- In Orbiter Processing Facility Bay 1, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) and United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (right) are briefed by a USA technician (center) on Shuttle processing in the payload bay of orbiter Atlantis. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- In Orbiter Processing Facility Bay 1, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) and United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (right) are briefed by a USA technician (center) on Shuttle processing in the payload bay of orbiter Atlantis. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro are briefed on the properties of the tile used in the Shuttle's Thermal Protection System (TPS) by USA Manager of the TPS Facility Martin Wilson (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro are briefed on the properties of the tile used in the Shuttle's Thermal Protection System (TPS) by USA Manager of the TPS Facility Martin Wilson (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Space Shuttle wind tunnel testing program

NASA Technical Reports Server (NTRS)

Whitnah, A. M.; Hillje, E. R.

1984-01-01

A major phase of the Space Shuttle Vehicle (SSV) Development Program was the acquisition of data through the space shuttle wind tunnel testing program. It became obvious that the large number of configuration/environment combinations would necessitate an extremely large wind tunnel testing program. To make the most efficient use of available test facilities and to assist the prime contractor for orbiter design and space shuttle vehicle integration, a unique management plan was devised for the design and development phase. The space shuttle program is reviewed together with the evolutional development of the shuttle configuration. The wind tunnel testing rationale and the associated test program management plan and its overall results is reviewed. Information is given for the various facilities and models used within this program. A unique posttest documentation procedure and a summary of the types of test per disciplines, per facility, and per model are presented with detailed listing of the posttest documentation.

Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells.

PubMed

Rosorius, O; Heger, P; Stelz, G; Hirschmann, N; Hauber, J; Stauber, R H

1999-08-01

We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For this purpose, substrates were created containing nuclear localization signals (NLS) or nuclear export signals (NES) linked to a chimeric protein composed of the glutathione S-transferase (GST) fused to the green fluorescent protein (GFP). The combination of GST/GFP-tagging allowed us to control protein expression in bacteria and to monitor protein purification during chromatography. Following microinjection into somatic cells, nuclear export/import of the highly fluorescent substrates could be observed directly by fluorescence microscopy. This system sets the stage to quantitate, in real time, the kinetics of nuclear import/export in living cells and to evaluate qualitative differences in various NLS/NES signals and pathways.

Arsenite promotes centrosome abnormalities under a p53 compromised status induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liao, W.-T.; Center of Excellence for Environmental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

2010-02-15

Epidemiological evidence indicated that residents, especially cigarette smokers, in arseniasis areas had significantly higher lung cancer risk than those living in non-arseniasis areas. Thus an interaction between arsenite and cigarette smoking in lung carcinogenesis was suspected. In the present study, we investigated the interactions of a tobacco-specific carcinogen 4- (methylnitrosamino)-1-(3-pyridyl)-1-butanone (nicotine-derived nitrosamine ketone, NNK) and arsenite on lung cell transformation. BEAS-2B, an immortalized human lung epithelial cell line, was selected to test the centrosomal abnormalities and colony formation by NNK and arsenite. We found that NNK, alone, could enhance BEAS-2B cell growth at 1-5 muM. Under NNK exposure, arsenite wasmore » able to increase centrosomal abnormality as compared with NNK or arsenite treatment alone. NNK treatment could also reduce arsenite-induced G2/M cell cycle arrest and apoptosis, these cellular effects were found to be correlated with p53 dysfunction. Increased anchorage-independent growth (colony formation) of BEAS-2B cells cotreated with NNK and arsenite was also observed in soft agar. Our present investigation demonstrated that NNK could provide a p53 compromised status. Arsenite would act specifically on this p53 compromised status to induce centrosomal abnormality and colony formation. These findings provided strong evidence on the carcinogenic promotional role of arsenite under tobacco-specific carcinogen co-exposure.« less

The conversion of centrioles to centrosomes: essential coupling of duplication with segregation

PubMed Central

Wang, Won-Jing; Soni, Rajesh Kumar; Uryu, Kunihiro

2011-01-01

Centrioles are self-reproducing organelles that form the core structure of centrosomes or microtubule-organizing centers (MTOCs). However, whether duplication and MTOC organization reflect innate activities of centrioles or activities acquired conditionally is unclear. In this paper, we show that newly formed full-length centrioles had no inherent capacity to duplicate or to organize pericentriolar material (PCM) but acquired both after mitosis through a Plk1-dependent modification that occurred in early mitosis. Modified centrioles initiated PCM recruitment in G1 and segregated equally in mitosis through association with spindle poles. Conversely, unmodified centrioles segregated randomly unless passively tethered to modified centrioles. Strikingly, duplication occurred only in centrioles that were both modified and disengaged, whereas unmodified centrioles, engaged or not, were “infertile,” indicating that engagement specifically blocks modified centrioles from reduplication. These two requirements, centriole modification and disengagement, fully exclude unlimited duplication in one cell cycle. We thus uncovered a Plk1-dependent mechanism whereby duplication and segregation are coupled to maintain centriole homeostasis. PMID:21576395

Space Shuttle mission: STS-67

NASA Technical Reports Server (NTRS)

1995-01-01

The Space Shuttle Endeavor, scheduled to launch March 2, 1995 from NASA's Kennedy Space Center, will conduct NASA's longest Shuttle flight prior to date. The mission, designated STS-67, has a number of experiments and payloads, which the crew, commanded by Stephen S. Oswald, will have to oversee. This NASA press kit for the mission contains a general background (general press release, media services information, quick-look facts page, shuttle abort modes, summary timeline, payload and vehicle weights, orbital summary, and crew responsibilities); cargo bay payloads and activities (Astro 2, Get Away Special Experiments); in-cabin payloads (Commercial Minimum Descent Altitude Instrumentation Technology Associates Experiments, protein crystal growth experiments, Middeck Active Control Experiment, and Shuttle Amateur Radio Experiment); and the STS-67 crew biographies. The payloads and experiments are described and summarized to give an overview of the goals, objectives, apparatuses, procedures, sponsoring parties, and the assigned crew members to carry out the tasks.

Autophagy mediates cell cycle response by regulating nucleocytoplasmic transport of PAX6 in limbal stem cells under ultraviolet-A stress

PubMed Central

Laggner, Maria; Pollreisz, Andreas; Schmidinger, Gerald; Schmidt-Erfurth, Ursula; Chen, Ying-Ting

2017-01-01

Limbal stem cells (LSC) account for homeostasis and regeneration of corneal epithelium. Solar ultraviolet A (UVA) is the major source causing oxidative damage in the ocular surface. Autophagy, a lysosomal degradation mechanism, is essential for physiologic function and stress defense of stem cells. PAX6, a master transcription factor governing corneal homeostasis by regulating cell cycle and cell fate of LSC, responds to oxidative stress by nucleocytoplasmic shuttling. Impaired autophagy and deregulated PAX6 have been reported in oxidative stress-related ocular surface disorders. We hypothesize a functional role for autophagy and PAX6 in LSC’s stress response to UVA. Therefore, human LSC colonies were irradiated with a sub-lethal dose of UVA and autophagic activity and intracellular reactive oxygen species (ROS) were measured by CYTO-ID assay and CM-H2DCFDA live staining, respectively. Following UVA irradiation, the percentage of autophagic cells significantly increased in LSC colonies while intracellular ROS levels remained unaffected. siRNA-mediated knockdown (KD) of ATG7 abolished UVA-induced autophagy and led to an excessive accumulation of ROS. Upon UVA exposure, LSCs displayed nuclear-to-cytoplasmic translocation of PAX6, while ATG7KD or antioxidant pretreatment largely attenuated the intracellular trafficking event. Immunofluorescence showing downregulation of proliferative marker PCNA and induction of cell cycle regulator p21 indicates cell cycle arrest in UVA-irradiated LSC. Abolishing autophagy, adenoviral-assisted restoration of nuclear PAX6 or antioxidant pretreatment abrogated the UVA-induced cell cycle arrest. Adenoviral expression of an ectopic PAX gene, PAX7, did not affect UVA cell cycle response. Furthermore, knocking down PAX6 attenuated the cell cycle progression of irradiated ATG7KD LSC by de-repressing p21 expression. Collectively, our data suggest a crosstalk between autophagy and PAX6 in regulating cell cycle response of ocular progenitors

Truncation of POC1A associated with short stature and extreme insulin resistance.

PubMed

Chen, Jian-Hua; Segni, Maria; Payne, Felicity; Huang-Doran, Isabel; Sleigh, Alison; Adams, Claire; Savage, David B; O'Rahilly, Stephen; Semple, Robert K; Barroso, Inês

2015-10-01

We describe a female proband with primordial dwarfism, skeletal dysplasia, facial dysmorphism, extreme dyslipidaemic insulin resistance and fatty liver associated with a novel homozygous frameshift mutation in POC1A, predicted to affect two of the three protein products of the gene. POC1A encodes a protein associated with centrioles throughout the cell cycle and implicated in both mitotic spindle and primary ciliary function. Three homozygous mutations affecting all isoforms of POC1A have recently been implicated in a similar syndrome of primordial dwarfism, although no detailed metabolic phenotypes were described. Primary cells from the proband we describe exhibited increased centrosome amplification and multipolar spindle formation during mitosis, but showed normal DNA content, arguing against mitotic skipping, cleavage failure or cell fusion. Despite evidence of increased DNA damage in cells with supernumerary centrosomes, no aneuploidy was detected. Extensive centrosome clustering both at mitotic spindles and in primary cilia mitigated the consequences of centrosome amplification, and primary ciliary formation was normal. Although further metabolic studies of patients with POC1A mutations are warranted, we suggest that POC1A may be added to ALMS1 and PCNT as examples of centrosomal or pericentriolar proteins whose dysfunction leads to extreme dyslipidaemic insulin resistance. Further investigation of links between these molecular defects and adipose tissue dysfunction is likely to yield insights into mechanisms of adipose tissue maintenance and regeneration that are critical to metabolic health. © 2015 Society for Endocrinology.

Shuttle program: Ground tracking data program document shuttle OFT launch/landing

NASA Technical Reports Server (NTRS)

Lear, W. M.

1977-01-01

The equations for processing ground tracking data during a space shuttle ascent or entry, or any nonfree flight phase of a shuttle mission are given. The resulting computer program processes data from up to three stations simultaneously: C-band station number 1; C-band station number 2; and an S-band station. The C-band data consists of range, azimuth, and elevation angle measurements. The S-band data consists of range, two angles, and integrated Doppler data in the form of cycle counts. A nineteen element state vector is used in Kalman filter to process the measurements. The acceleration components of the shuttle are taken to be independent exponentially-correlated random variables. Nine elements of the state vector are the measurement bias errors associated with range and two angles for each tracking station. The biases are all modeled as exponentially-correlated random variables with a typical time constant of 108 seconds. All time constants are taken to be the same for all nine state variables. This simplifies the logic in propagating the state error covariance matrix ahead in time.

Infrared emission associated with chemical reactions on Shuttle and SIRTF surfaces

NASA Technical Reports Server (NTRS)

Hollenbach, D. J.; Tielens, Alexander G. G. M.

1984-01-01

The infrared intensities which would be observed by the Shuttle Infrared Telescope Facility (SIRTF), and which are produced by surface chemistry following atmospheric impact on SIRTF and the shuttle are estimated. Three possible sources of reactants are analyzed: (1) direct atmospheric and scattered contaminant fluxes onto the shuttle's surface; (2) direct atmospheric and scattered contaminant fluxes onto the SIRTF sunshade; and (3) scattered fluxes onto the cold SIRTF mirror. The chemical reactions are primarily initiated by the dominent flux of reactive atomic oxygen on the surfaces. Using observations of the optical glow to constrain theoretical parameters, it is estimated for source (1) that the infrared glow on the SIRTF mirror will be comparable to the zodiacal background between 1 and 10 micron wavelengths. It is speculated that oxygen reacts with the atoms and the radicals bound in the organic molecules that reside on the shuttle and the Explorer surfaces. It is concluded that for source (2) that with suitable construction, a warm sunshade will produce insignificant infrared glow. It is noted that the atomic oxygen flux on the cold SIRTF mirror (3) is insufficient to produce significant infrared glow. Infrared absorption by the ice buildup on the mirror is also small.

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Manager of the Thermal Protection System (TPS) Facility Martin Wilson (right) briefs NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) on the properties of a thermal blanket used in the Shuttle's TPS. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Manager of the Thermal Protection System (TPS) Facility Martin Wilson (right) briefs NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) on the properties of a thermal blanket used in the Shuttle's TPS. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) discusses some of the working parts inside the nose of Shuttle Discovery in Orbiter Processing Facility Bay 3 with a United Space Alliance (USA) technician (back to camera). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) discusses some of the working parts inside the nose of Shuttle Discovery in Orbiter Processing Facility Bay 3 with a United Space Alliance (USA) technician (back to camera). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

KENNEDY SPACE CENTER, FLA. -- From front row left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and NASA Space Shuttle Program Manager William Parsons are trained on the proper use of the Emergency Life Support Apparatus (ELSA). NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From front row left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and NASA Space Shuttle Program Manager William Parsons are trained on the proper use of the Emergency Life Support Apparatus (ELSA). NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

STS-121 Space Shuttle Processing Update

NASA Image and Video Library

2006-04-27

NASA Administrator Michael Griffin, left, and Associate Administrator for Space Operations William Gerstenmaier, right, look on as Space Shuttle Program Manager Wayne Hale from NASA's Marshall Space Flight Center, holds a test configuration of an ice frost ramp during a media briefing about the space shuttle program and processing for the STS-121 mission, Friday, April 28, 2006, at NASA Headquarters in Washington. Photo Credit (NASA/Bill Ingalls)

Transforming growth factor beta-independent shuttling of Smad4 between the cytoplasm and nucleus.

PubMed

Pierreux, C E; Nicolás, F J; Hill, C S

2000-12-01

Smad4 plays a pivotal role in all transforming growth factor beta (TGF-beta) signaling pathways. Here we describe six widely expressed alternatively spliced variants of human Smad4 with deletions of different exons in the linker, the region of Smad4 that separates the two well-conserved MH1 and MH2 domains. All these Smad4 variants form complexes with activated Smad2 and Smad3 and are incorporated into DNA-binding complexes with the transcription factor Fast-1, regardless of the amount of linker they contain. However, sequences encoded by exons 5 to 7 in the linker are essential for transcriptional activation. Most importantly, our observation that different Smad4 isoforms have different subcellular localizations has led us to the identification of a functional CRM1-dependent nuclear export signal in the Smad4 linker and a constitutively active nuclear localization signal in the N-terminal MH1 domain. In the absence of TGF-beta signaling, we conclude that Smad4 is rapidly and continuously shuttling between the nucleus and the cytoplasm, the distribution of Smad4 between the nucleus and the cytoplasm being dictated by the relative strengths of the nuclear import and export signals. We demonstrate that inhibition of CRM1-mediated nuclear export by treatment of cells with leptomycin B results in endogenous Smad4 accumulating very rapidly in the nucleus. Endogenous Smad2 and Smad3 are completely unaffected by leptomycin B treatment, indicating that the nucleocytoplasmic shuttling is specific for Smad4. We propose that, upon TGF-beta signaling, complex formation between Smad4 and activated Smad2 or -3 leads to nuclear accumulation of Smad4 through inhibition of its nuclear export. We demonstrate that after prolonged TGF-beta signaling Smad2 becomes dephosphorylated and Smad2 and Smad4 accumulate back in the cytoplasm.

Transforming Growth Factor β-Independent Shuttling of Smad4 between the Cytoplasm and Nucleus

PubMed Central

Pierreux, Christophe E.; Nicolás, Francisco J.; Hill, Caroline S.

2000-01-01

Smad4 plays a pivotal role in all transforming growth factor β (TGF-β) signaling pathways. Here we describe six widely expressed alternatively spliced variants of human Smad4 with deletions of different exons in the linker, the region of Smad4 that separates the two well-conserved MH1 and MH2 domains. All these Smad4 variants form complexes with activated Smad2 and Smad3 and are incorporated into DNA-binding complexes with the transcription factor Fast-1, regardless of the amount of linker they contain. However, sequences encoded by exons 5 to 7 in the linker are essential for transcriptional activation. Most importantly, our observation that different Smad4 isoforms have different subcellular localizations has led us to the identification of a functional CRM1-dependent nuclear export signal in the Smad4 linker and a constitutively active nuclear localization signal in the N-terminal MH1 domain. In the absence of TGF-β signaling, we conclude that Smad4 is rapidly and continuously shuttling between the nucleus and the cytoplasm, the distribution of Smad4 between the nucleus and the cytoplasm being dictated by the relative strengths of the nuclear import and export signals. We demonstrate that inhibition of CRM1-mediated nuclear export by treatment of cells with leptomycin B results in endogenous Smad4 accumulating very rapidly in the nucleus. Endogenous Smad2 and Smad3 are completely unaffected by leptomycin B treatment, indicating that the nucleocytoplasmic shuttling is specific for Smad4. We propose that, upon TGF-β signaling, complex formation between Smad4 and activated Smad2 or -3 leads to nuclear accumulation of Smad4 through inhibition of its nuclear export. We demonstrate that after prolonged TGF-β signaling Smad2 becomes dephosphorylated and Smad2 and Smad4 accumulate back in the cytoplasm. PMID:11074002

Viral Proteinase Requirements for the Nucleocytoplasmic Relocalization of Cellular Splicing Factor SRp20 during Picornavirus Infections

PubMed Central

Fitzgerald, Kerry D.; Chase, Amanda J.; Cathcart, Andrea L.; Tran, Genevieve P.

2013-01-01

Infection of mammalian cells by picornaviruses results in the nucleocytoplasmic redistribution of certain host cell proteins. These viruses interfere with import-export pathways, allowing for the cytoplasmic accumulation of nuclear proteins that are then available to function in viral processes. We recently described the cytoplasmic relocalization of cellular splicing factor SRp20 during poliovirus infection. SRp20 is an important internal ribosome entry site (IRES) trans-acting factor (ITAF) for poliovirus IRES-mediated translation; however, it is not known whether other picornaviruses utilize SRp20 as an ITAF and direct its cytoplasmic relocalization. Also, the mechanism by which poliovirus directs the accumulation of SRp20 in the cytoplasm of the infected cell is currently unknown. Work described in this report demonstrated that infection by another picornavirus (coxsackievirus B3) causes SRp20 to relocalize from the nucleus to the cytoplasm of HeLa cells, similar to poliovirus infection; however, SRp20 is relocalized to a somewhat lesser extent in the cytoplasm of HeLa cells during infection by yet another picornavirus (human rhinovirus 16). We show that expression of poliovirus 2A proteinase is sufficient to cause the nucleocytoplasmic redistribution of SRp20. Following expression of poliovirus 2A proteinase in HeLa cells, we detect cleavage of specific nuclear pore proteins known to be cleaved during poliovirus infection. We also find that expression of human rhinovirus 16 2A proteinase alone can cause efficient cytoplasmic relocalization of SRp20, despite the lower levels of SRp20 relocalization observed during rhinovirus infection compared to poliovirus. Taken together, these results further define the mechanism of SRp20 cellular redistribution during picornavirus infections, and they provide additional insight into some of the differences observed between human rhinovirus and other enterovirus infections. PMID:23255796

Shuttle/GPSPAC experimentation study

NASA Technical Reports Server (NTRS)

Moses, J.; Flack, J. F.

1977-01-01

The utilization is discussed of the GPSPAC, which is presently being developed to be used on the low altitude host vehicle (LAHV), for possible use in the shuttle avionics system to evaluate shuttle/GPS navigation performance. Analysis and tradeoffs of the shuttle/GPS link, shuttle signal interface requirements, oscillator tradeoffs and GPSPAC mechanical modifications for shuttle are included. Only the on-orbit utilization of GPSPAC for the shuttle is discussed. Other phases are briefly touched upon. Recommendations are provided for using the present GPSPAC and the changes required to perform shuttle on-orbit navigation.

Nucleocytoplasmic shuttling of hexokinase II in a cancer cell

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neary, Catherine L., E-mail: nearycl@umdnj.edu; Pastorino, John G.

2010-04-16

In yeast, the hexokinase type II enzyme (HXKII) translocates to the nucleus in the presence of excess glucose, and participates in glucose repression. However, no evidence has suggested a nuclear function for HXKII in mammalian cells. Herein, we present data showing nuclear localization of HXKII in HeLa cells, both by immunocytochemistry and subcellular fractionation. HXKII is extruded from the nucleus, at least in part, by the activity of the exportin 1/CrmA system, as demonstrated by increased nuclear expression and decreased cytoplasmic expression after incubation with leptomycin B, a bacterially-derived exportin inhibitor. Furthermore, cytoplasmic localization of HXKII is dependent on itsmore » enzymatic activity, as inhibiting HXKII activity using 2-deoxy-D-glucose (2DG) increased nuclear localization. This effect was more significant in cells incubated in the absence of glucose for 24 h prior to addition of 2DG. Regulated translocation of HXKII to the nucleus of mammalian cells could represent a previously unknown glucose-sensing mechanism.« less

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Associate Program Manager of Florida Operations Bill Pickavance (left front) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right front) tour a solid rocket booster (SRB) retrieval ship at Cape Canaveral. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Associate Program Manager of Florida Operations Bill Pickavance (left front) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (right front) tour a solid rocket booster (SRB) retrieval ship at Cape Canaveral. NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

FOOD - SHUTTLE

NASA Image and Video Library

1982-02-01

S82-26423 (January 1982) --- This is a close-up view of the rehydration unit to be used in meal preparation on operational space shuttle flights. The unit is located on the middeck of the space shuttles in the NASA fleet. Note the part of the food tray in upper right corner. Its six compartments (not all pictured) are used in space shuttle meal preparation and consumption. Photo credit: NASA

Shuttle Safety Improvements

NASA Technical Reports Server (NTRS)

Henderson, Edward

2001-01-01

The Space Shuttle has been flying for over 20 years and based on the Orbiter design life of 100 missions it should be capable of flying at least 20 years more if we take care of it. The Space Shuttle Development Office established in 1997 has identified those upgrades needed to keep the Shuttle flying safely and efficiently until a new reusable launch vehicle (RLV) is available to meet the agency commitments and goals for human access to space. The upgrade requirements shown in figure 1 are to meet the program goals, support HEDS and next generation space transportation goals while protecting the country 's investment in the Space Shuttle. A major review of the shuttle hardware and processes was conducted in 1999 which identified key shuttle safety improvement priorities, as well as other system upgrades needed to reliably continue to support the shuttle miss ions well into the second decade of this century. The high priority safety upgrades selected for development and study will be addressed in this paper.

The Shuttle Era

NASA Technical Reports Server (NTRS)

1981-01-01

An overview of the Space Shuttle Program is presented. The missions of the space shuttle orbiters, the boosters and main engine, and experimental equipment are described. Crew and passenger accommodations are discussed as well as the shuttle management teams.

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and NASA Space Shuttle Program Manager William Parsons each don an Emergency Life Support Apparatus (ELSA) during training on the proper use of the escape devices. NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- From left, NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik and NASA Space Shuttle Program Manager William Parsons each don an Emergency Life Support Apparatus (ELSA) during training on the proper use of the escape devices. NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Shuttle Carrier Aircraft

NASA Image and Video Library

2014-04-23

It has been called the world's greatest piggyback ride: a space shuttle, atop a Boeing 747 jet aircraft. But this is no ordinary 747, this is the Shuttle Carrier Aircraft...the SCA. This specially modified jumbo jet was not only a taxi service for the shuttle, but also helped in the development of the shuttle itself. In 30 years of flying, the majestic image of a spacecraft joined to the SCA, became a symbol of American invention and ingenuity.

Autonomous Space Shuttle

NASA Technical Reports Server (NTRS)

Siders, Jeffrey A.; Smith, Robert H.

2004-01-01

The continued assembly and operation of the International Space Station (ISS) is the cornerstone within NASA's overall Strategic P an. As indicated in NASA's Integrated Space Transportation Plan (ISTP), the International Space Station requires Shuttle to fly through at least the middle of the next decade to complete assembly of the Station, provide crew transport, and to provide heavy lift up and down mass capability. The ISTP reflects a tight coupling among the Station, Shuttle, and OSP programs to support our Nation's space goal . While the Shuttle is a critical component of this ISTP, there is a new emphasis for the need to achieve greater efficiency and safety in transporting crews to and from the Space Station. This need is being addressed through the Orbital Space Plane (OSP) Program. However, the OSP is being designed to "complement" the Shuttle as the primary means for crew transfer, and will not replace all the Shuttle's capabilities. The unique heavy lift capabilities of the Space Shuttle is essential for both ISS, as well as other potential missions extending beyond low Earth orbit. One concept under discussion to better fulfill this role of a heavy lift carrier, is the transformation of the Shuttle to an "un-piloted" autonomous system. This concept would eliminate the loss of crew risk, while providing a substantial increase in payload to orbit capability. Using the guidelines reflected in the NASA ISTP, the autonomous Shuttle a simplified concept of operations can be described as; "a re-supply of cargo to the ISS through the use of an un-piloted Shuttle vehicle from launch through landing". Although this is the primary mission profile, the other major consideration in developing an autonomous Shuttle is maintaining a crew transportation capability to ISS as an assured human access to space capability.

Space Shuttle Projects

NASA Image and Video Library

2002-03-07

STS-109 Astronaut Michael J. Massimino, mission specialist, perched on the Shuttle's robotic arm is working at the stowage area for the Hubble Space Telescope's port side solar array. Working in tandem with James. H. Newman, Massimino removed the old port solar array and stored it in Columbia's payload bay for return to Earth. The two went on to install a third generation solar array and its associated electrical components. Two crew mates had accomplished the same feat with the starboard array on the previous day. In addition to the replacement of the solar arrays, the STS-109 crew also installed the experimental cooling system for the Hubble's Near-Infrared Camera (NICMOS), replaced the power control unit (PCU), and replaced the Faint Object Camera (FOC) with a new advanced camera for Surveys (ACS). The 108th flight overall in NASA's Space Shuttle Program, the Space Shuttle Columbia STS-109 mission lifted off March 1, 2002 for 10 days, 22 hours, and 11 minutes. Five space walks were conducted to complete the HST upgrades. The Marshall Space Flight Center in Huntsville, Alabama had the responsibility for the design, development, and construction of the HST, which is the most powerful and sophisticated telescope ever built.

Space Shuttle Projects

NASA Image and Video Library

2001-01-01

The Space Shuttle represented an entirely new generation of space vehicles, the world's first reusable spacecraft. Unlike earlier expendable rockets, the Shuttle was designed to be launched over and over again and would serve as a system for ferrying payloads and persornel to and from Earth orbit. The Shuttle's major components are the orbiter spacecraft; the three main engines, with a combined thrust of more than 1.2 million pounds; the huge external tank (ET) that feeds the liquid hydrogen fuel and liquid oxygen oxidizer to the three main engines; and the two solid rocket boosters (SRB's), with their combined thrust of some 5.8 million pounds, that provide most of the power for the first two minutes of flight. Crucially involved with the Space Shuttle program virtually from its inception, the Marshall Space Flight Center (MSFC) played a leading role in the design, development, testing, and fabrication of many major Shuttle propulsion components. The MSFC was assigned responsibility for developing the Shuttle orbiter's high-performance main engines, the most complex rocket engines ever built. The MSFC was also responsible for developing the Shuttle's massive ET and the solid rocket motors and boosters.

Space Shuttle Projects

NASA Image and Video Library

1975-01-01

The Space Shuttle represented an entirely new generation of space vehicle, the world's first reusable spacecraft. Unlike earlier expendable rockets, the Shuttle was designed to be launched over and over again and would serve as a system for ferrying payloads and persornel to and from Earth orbit. The Shuttle's major components are the orbiter spacecraft; the three main engines, with a combined thrust of more than 1.2 million pounds; the huge external tank (ET) that feeds the liquid hydrogen fuel and liquid oxygen oxidizer to the three main engines; and the two solid rocket boosters (SRB's), with their combined thrust of some 5.8 million pounds. The SRB's provide most of the power for the first two minutes of flight. Crucially involved with the Space Shuttle program virtually from its inception, the Marshall Space Flight Center (MSFC) played a leading role in the design, development, testing, and fabrication of many major Shuttle propulsion components. The MSFC was assigned responsibility for developing the Shuttle orbiter's high-performance main engines, the most complex rocket engines ever built. The MSFC was also responsible for developing the Shuttle's massive ET and the solid rocket motors and boosters.

Post-Shuttle EVA Operations on ISS

NASA Technical Reports Server (NTRS)

West, Bill; Witt, Vincent; Chullen, Cinda

2010-01-01

The EVA hardware used to assemble and maintain the ISS was designed with the assumption that it would be returned to Earth on the Space Shuttle for ground processing, refurbishment, or failure investigation (if necessary). With the retirement of the Space Shuttle, a new concept of operations was developed to enable EVA hardware (EMU, Airlock Systems, EVA tools, and associated support equipment and consumables) to perform ISS EVAs until 2016 and possibly beyond to 2020. Shortly after the decision to retire the Space Shuttle was announced, NASA and the One EVA contractor team jointly initiated the EVA 2010 Project. Challenges were addressed to extend the operating life and certification of EVA hardware, secure the capability to launch EVA hardware safely on alternate launch vehicles, and protect EMU hardware operability on orbit for long durations.

STS-37 Space Shuttle mission report

NASA Astrophysics Data System (ADS)

Fricke, Robert W.

1991-05-01

The STS-37 Space Shuttle Program Mission Report contains a summary of the vehicle subsystem activities during this thirty-ninth flight of the Space Shuttle and the eighth flight of the Orbiter Vehicle Atlantis (OV-104). In addition to the Atlantis vehicle, the flight vehicle consisted of the following: an External Tank (ET) (designated as ET-37/LWT-30); three Space Shuttle main engines (SSME's) (serial numbers 2019, 2031, and 2107 in positions 1, 2, and 3, respectively); and two Solid Rocket Boosters (SRB's) designated as BI-042. The primary objective of this flight was to successfully deploy the Gamma Ray Observatory (GRO) payload. The secondary objectives were to successfully perform all operations necessary to support the requirements of the Protein Crystal Growth (PCG) Block 2 version, Radiation Monitoring Experiment-3 (RME-3), Ascent Particle Monitor (APM), Shuttle Amateur Radio Experiment-2 (SAREX-2), Air Force Maui Optical Site Calibration Test (AMOS), Bioserve Instrumentation Technology Associates Materials Dispersion Apparatus (BIMDA), and the Crew and Equipment Transfer Aids (CETA) payloads.

STS-37 Space Shuttle mission report

NASA Technical Reports Server (NTRS)

Fricke, Robert W.

1991-01-01

The STS-37 Space Shuttle Program Mission Report contains a summary of the vehicle subsystem activities during this thirty-ninth flight of the Space Shuttle and the eighth flight of the Orbiter Vehicle Atlantis (OV-104). In addition to the Atlantis vehicle, the flight vehicle consisted of the following: an External Tank (ET) (designated as ET-37/LWT-30); three Space Shuttle main engines (SSME's) (serial numbers 2019, 2031, and 2107 in positions 1, 2, and 3, respectively); and two Solid Rocket Boosters (SRB's) designated as BI-042. The primary objective of this flight was to successfully deploy the Gamma Ray Observatory (GRO) payload. The secondary objectives were to successfully perform all operations necessary to support the requirements of the Protein Crystal Growth (PCG) Block 2 version, Radiation Monitoring Experiment-3 (RME-3), Ascent Particle Monitor (APM), Shuttle Amateur Radio Experiment-2 (SAREX-2), Air Force Maui Optical Site Calibration Test (AMOS), Bioserve Instrumentation Technology Associates Materials Dispersion Apparatus (BIMDA), and the Crew and Equipment Transfer Aids (CETA) payloads.

A molecular mechanism of mitotic centrosome assembly in Drosophila

PubMed Central

Conduit, Paul T; Richens, Jennifer H; Wainman, Alan; Holder, James; Vicente, Catarina C; Pratt, Metta B; Dix, Carly I; Novak, Zsofia A; Dobbie, Ian M; Schermelleh, Lothar; Raff, Jordan W

2014-01-01

Centrosomes comprise a pair of centrioles surrounded by pericentriolar material (PCM). The PCM expands dramatically as cells enter mitosis, but it is unclear how this occurs. In this study, we show that the centriole protein Asl initiates the recruitment of DSpd-2 and Cnn to mother centrioles; both proteins then assemble into co-dependent scaffold-like structures that spread outwards from the mother centriole and recruit most, if not all, other PCM components. In the absence of either DSpd-2 or Cnn, mitotic PCM assembly is diminished; in the absence of both proteins, it appears to be abolished. We show that DSpd-2 helps incorporate Cnn into the PCM and that Cnn then helps maintain DSpd-2 within the PCM, creating a positive feedback loop that promotes robust PCM expansion around the mother centriole during mitosis. These observations suggest a surprisingly simple mechanism of mitotic PCM assembly in flies. DOI: http://dx.doi.org/10.7554/eLife.03399.001 PMID:25149451

Search, capture and signal: games microtubules and centrosomes play.

PubMed

Schuyler, S C; Pellman, D

2001-01-01

Accurate distribution of the chromosomes in dividing cells requires coupling of cellular polarity cues with both the orientation of the mitotic spindle and cell cycle progression. Work in budding yeast has demonstrated that cytoplasmic dynein and the kinesin Kip3p define redundant pathways that ensure proper spindle orientation. Furthermore, it has been shown that the Kip3p pathway components Kar9p and Bim1p (Yeb1p) form a complex that provides a molecular link between cortical polarity cues and spindle microtubules. Recently, other studies indicated that the cortical localization of Kar9p depends upon actin cables and Myo2p, a type V myosin. In addition, a BUB2-dependent cell cycle checkpoint has been described that inhibits the mitotic exit network and cytokinesis until proper centrosome position is achieved. Combined, these studies provide molecular insight into how cells link cellular polarity, spindle position and cell cycle progression.

STS-63 Space Shuttle report

NASA Technical Reports Server (NTRS)

Fricke, Robert W., Jr.

1995-01-01

The STS-63 Space Shuttle Program Mission Report summarizes the Payload activities and provides detailed data on the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Reusable Solid Rocket Motor (RSRM), and the Space Shuttle Main Engine (SSME) systems performance during this sixty-seventh flight of the Space Shuttle Program, the forty-second since the return to flight, and twentieth flight of the Orbiter vehicle Discovery (OV-103). In addition to the OV-103 Orbiter vehicle, the flight vehicle consisted of an ET that was designated ET-68; three SSME's that were designated 2035, 2109, and 2029 in positions 1, 2, and 3, respectively; and two SRB's that were designated BI-070. The RSRM's that were an integral part of the SRB's were designated 360Q042A for the left SRB and 360L042B for the right SRB. The STS-63 mission was planned as an 8-day duration mission with two contingency days available for weather avoidance or Orbiter contingency operations. The primary objectives of the STS-63 mission were to perform the Mir rendezvous operations, accomplish the Spacehab-3 experiments, and deploy and retrieve the Shuttle Pointed Autonomous Research Tool for Astronomy-204 (SPARTAN-204) payload. The secondary objectives were to perform the Cryogenic Systems Experiment (CSE)/Shuttle Glo-2 Experiment (GLO-2) Payload (CGP)/Orbital Debris Radar Calibration Spheres (ODERACS-2) (CGP/ODERACS-2) payload objectives, the Solid Surface Combustion Experiment (SSCE), and the Air Force Maui Optical Site Calibration Tests (AMOS). The objectives of the Mir rendezvous/flyby were to verify flight techniques, communication and navigation-aid sensor interfaces, and engineering analyses associated with Shuttle/Mir proximity operations in preparation for the STS-71 docking mission.

Space Shuttle news reference

NASA Technical Reports Server (NTRS)

1981-01-01

A detailed description of the space shuttle vehicle and associated subsystems is given. Space transportation system propulsion, power generation, environmental control and life support system and avionics are among the topics. Also, orbiter crew accommodations and equipment, mission operations and support, and flight crew complement and crew training are addressed.

Differential sensitivity of Glioma stem cells to Aurora kinase A inhibitors: implications for stem cell mitosis and centrosome dynamics.

PubMed

Mannino, Mariella; Gomez-Roman, Natividad; Hochegger, Helfrid; Chalmers, Anthony J

2014-07-01

Glioma stem-cell-like cells are considered to be responsible for treatment resistance and tumour recurrence following chemo-radiation in glioblastoma patients, but specific targets by which to kill the cancer stem cell population remain elusive. A characteristic feature of stem cells is their ability to undergo both symmetric and asymmetric cell divisions. In this study we have analysed specific features of glioma stem cell mitosis. We found that glioma stem cells appear to be highly prone to undergo aberrant cell division and polyploidization. Moreover, we discovered a pronounced change in the dynamic of mitotic centrosome maturation in these cells. Accordingly, glioma stem cell survival appeared to be strongly dependent on Aurora A activity. Unlike differentiated cells, glioma stem cells responded to moderate Aurora A inhibition with spindle defects, polyploidization and a dramatic increase in cellular senescence, and were selectively sensitive to Aurora A and Plk1 inhibitor treatment. Our study proposes inhibition of centrosomal kinases as a novel strategy to selectively target glioma stem cells. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) tours a solid rocket booster (SRB) retrieval ship at Cape Canaveral. NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (left) tours a solid rocket booster (SRB) retrieval ship at Cape Canaveral. NASA and United Space Alliance (USA) Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Space shuttle requirements/configuration evolution

NASA Technical Reports Server (NTRS)

Andrews, E. P.

1991-01-01

Space Shuttle chronology; Space Shuttle comparison; Cost comparison; Performance; Program ground rules; Sizing criteria; Crew/passenger provisions; Space Shuttle Main Engine (SSME) characteristics; Space Shuttle program milestones; and Space Shuttle requirements are outlined. This presentation is represented by viewgraphs.

DISC1, PDE4B, and NDE1 at the centrosome and synapse

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bradshaw, Nicholas J.; Ogawa, Fumiaki; Antolin-Fontes, Beatriz

Disrupted-In-Schizophrenia 1 (DISC1) is a risk factor for schizophrenia and other major mental illnesses. Its protein binding partners include the Nuclear Distribution Factor E Homologs (NDE1 and NDEL1), LIS1, and phosphodiesterases 4B and 4D (PDE4B and PDE4D). We demonstrate that NDE1, NDEL1 and LIS1, together with their binding partner dynein, associate with DISC1, PDE4B and PDE4D within the cell, and provide evidence that this complex is present at the centrosome. LIS1 and NDEL1 have been previously suggested to be synaptic, and we now demonstrate localisation of DISC1, NDE1, and PDE4B at synapses in cultured neurons. NDE1 is phosphorylated by cAMP-dependantmore » Protein Kinase A (PKA), whose activity is, in turn, regulated by the cAMP hydrolysis activity of phosphodiesterases, including PDE4. We propose that DISC1 acts as an assembly scaffold for all of these proteins and that the NDE1/NDEL1/LIS1/dynein complex is modulated by cAMP levels via PKA and PDE4.« less

Hormone- and light-regulated nucleocytoplasmic transport in plants: current status.

PubMed

Lee, Yew; Lee, Hak-Soo; Lee, June-Seung; Kim, Seong-Ki; Kim, Soo-Hwan

2008-01-01

The gene regulation mechanisms underlying hormone- and light-induced signal transduction in plants rely not only on post-translational modification and protein degradation, but also on selective inclusion and exclusion of proteins from the nucleus. For example, plant cells treated with light or hormones actively transport many signalling regulatory proteins, transcription factors, and even photoreceptors and hormone receptors into the nucleus, while actively excluding other proteins. The nuclear envelope (NE) is the physical and functional barrier that mediates this selective partitioning, and nuclear transport regulators transduce hormone- or light-initiated signalling pathways across the membrane to mediate nuclear activities. Recent reports revealed that mutating the proteins regulating nuclear transport through the pores, such as nucleoporins, alters the plant's response to a stimulus. In this review, recent works are introduced that have revealed the importance of regulated nucleocytoplasmic partitioning. These important findings deepen our understanding about how co-ordinated plant hormone and light signal transduction pathways facilitate communication between the cytoplasm and the nucleus. The roles of nucleoporin components within the nuclear pore complex (NPC) are also emphasized, as well as nuclear transport cargo, such as Ran/TC4 and its binding proteins (RanBPs), in this process. Recent findings concerning these proteins may provide a possible direction by which to characterize the regulatory potential of hormone- or light-triggered nuclear transport.

ITOS/space shuttle study

NASA Technical Reports Server (NTRS)

1971-01-01

The results are reported of a study to explore the potential cost reductions in the operational ITOS weather satellite program as a consequence of shuttle/bug availability for satellite placement and retrieval, and satellite servicing and maintenance. The study program was divided into shuttle impact on equipment and testing costs, and shuttle impact on overall future ITOS operational program costs, and shuttle impact on configuration. It is concluded that savings in recurring spacecraft costs can be realized in the 1978 ITOS program, if a space shuttle is utilized.

Shuttle plate braiding machine

NASA Technical Reports Server (NTRS)

Huey, Jr., Cecil O. (Inventor)

1994-01-01

A method and apparatus for moving yarn in a selected pattern to form a braided article. The apparatus includes a segmented grid of stationary support elements and a plurality of shuttles configured to carry yarn. The shuttles are supported for movement on the grid assembly and each shuttle includes a retractable plunger for engaging a reciprocating shuttle plate that moves below the grid assembly. Such engagement at selected times causes the shuttles to move about the grid assembly in a selected pattern to form a braided article of a particular geometry.

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (center) are briefed on the use of a cold plate in Orbiter Processing Facility Bay 2 by a USA technician (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- United Space Alliance (USA) Vice President and Space Shuttle Program Manager Howard DeCastro (left) and NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (center) are briefed on the use of a cold plate in Orbiter Processing Facility Bay 2 by a USA technician (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Shuttle Carrier Aircraft (SCA) Fleet Photo

NASA Technical Reports Server (NTRS)

1995-01-01

NASA's two Boeing 747 Shuttle Carrier Aircraft (SCA) are seen here nose to nose at Dryden Flight Research Center, Edwards, California. The front mounting attachment for the Shuttle can just be seen on top of each. The SCAs are used to ferry Space Shuttle orbiters from landing sites back to the launch complex at the Kennedy Space Center, and also to and from other locations too distant for the orbiters to be delivered by ground transportation. The orbiters are placed atop the SCAs by Mate-Demate Devices, large gantry-like structures which hoist the orbiters off the ground for post-flight servicing, and then mate them with the SCAs for ferry flights. Features which distinguish the two SCAs from standard 747 jetliners are; three struts, with associated interior structural strengthening, protruding from the top of the fuselage (two aft, one forward) on which the orbiter is attached, and two additional vertical stabilizers, one on each end of the standard horizontal stabilizer, to enhance directional stability. The two SCAs are under the operational control of NASA's Johnson Space Center, Houston, Texas. Space Shuttles are the main element of America's Space Transportation System and are used for space research and other space applications. The shuttles are the first vehicles capable of being launched into space and returning to Earth on a routine basis. Space Shuttles are used as orbiting laboratories in which scientists and mission specialists conduct a wide variety of scientific experiments. Crews aboard shuttles place satellites in orbit, rendezvous with satellites to carry out repair missions and return them to space, and retrieve satellites and return them to Earth for refurbishment and reuse. Space Shuttles are true aerospace vehicles. They leave Earth and its atmosphere under rocket power provided by three liquid-propellant main engines with two solid-propellant boosters attached plus an external liquid-fuel tank. After their orbital missions, they streak back

Nucleocytoplasmic protein translocation during mitosis in the social amoebozoan Dictyostelium discoideum.

PubMed

O'Day, Danton H; Budniak, Aldona

2015-02-01

Mitosis is a fundamental and essential life process. It underlies the duplication and survival of all cells and, as a result, all eukaryotic organisms. Since uncontrolled mitosis is a dreaded component of many cancers, a full understanding of the process is critical. Evolution has led to the existence of three types of mitosis: closed, open, and semi-open. The significance of these different mitotic species, how they can lead to a full understanding of the critical events that underlie the asexual duplication of all cells, and how they may generate new insights into controlling unregulated cell division remains to be determined. The eukaryotic microbe Dictyostelium discoideum has proved to be a valuable biomedical model organism. While it appears to utilize closed mitosis, a review of the literature suggests that it possesses a form of mitosis that lies in the middle between truly open and fully closed mitosis-it utilizes a form of semi-open mitosis. Here, the nucleocytoplasmic translocation patterns of the proteins that have been studied during mitosis in the social amoebozoan D. discoideum are detailed followed by a discussion of how some of them provide support for the hypothesis of semi-open mitosis. © 2014 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.

Modulation of integrin-linked kinase nucleo-cytoplasmic shuttling by ILKAP and CRM1.

PubMed

Nakrieko, Kerry-Ann; Vespa, Alisa; Mason, David; Irvine, Timothy S; D'Souza, Sudhir J A; Dagnino, Lina

2008-07-15

Integrin-linked kinase (ILK) plays key roles in a variety of cell functions, including cell proliferation, adhesion and migration. Within the cell, ILK localizes to multiple sites, including the cytoplasm, focal adhesion complexes that mediate cell adhesion to extracellular substrates, as well as cell-cell junctions in epidermal keratinocytes. Central to understanding ILK function is the elucidation of the mechanisms that regulate its subcellular localization. We now demonstrate that ILK is imported into the nucleus through sequences in its N-terminus, via active transport mechanisms that involve nuclear pore complexes. In addition, nuclear ILK can be rapidly exported into the cytoplasm through a CRM1-dependent pathway, and its export is enhanced by the type 2C protein phosphatase ILKAP. Nuclear localization of ILK in epidermal keratinocytes is associated with increased DNA synthesis, which is sensitive to inhibition by ILKAP. Our studies demonstrate the importance for keratinocyte proliferation of ILK regulation through changes in its subcellular localization, and establish ILKAP and CRM1 as pivotal modulators of ILK subcellular distribution and activity in these cells.

Cardiovirus Leader proteins bind exportins: Implications for virus replication and nucleocytoplasmic trafficking inhibition

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ciomperlik, Jessica J.; Basta, Holly A.; Palmenberg, Ann C., E-mail: acpalmen@wisc.edu

2016-01-15

Cardiovirus Leader proteins (L{sub X}) inhibit cellular nucleocytoplasmic trafficking by directing host kinases to phosphorylate Phe/Gly-containing nuclear pore proteins (Nups). Resolution of the Mengovirus L{sub M} structure bound to Ran GTPase, suggested this complex would further recruit specific exportins (karyopherins), which in turn mediate kinase selection. Pull-down experiments and recombinant complex reconstitution now confirm that Crm1 and CAS exportins form stable dimeric complexes with encephalomyocarditis virus L{sub E}, and also larger complexes with L{sub E}:Ran. shRNA knockdown studies support this idea. Similar activities could be demonstrated for recombinant L{sub S} and L{sub T} from Theiloviruses. When mutations were introduced tomore » alter the L{sub E} zinc finger domain, acidic domain, or dual phosphorylation sites, there was reduced exportin selection. These regions are not involved in Ran interactions, so the Ran and Crm1 binding sites on L{sub E} must be non-overlapping. The involvement of exportins in this mechanism is important to viral replication and the observation of trafficking inhibition by L{sub E}.« less

Space Shuttle Pinhole Formation Mechanism Studies

NASA Technical Reports Server (NTRS)

Jacobson, Nathan S.

1998-01-01

Pinholes have been observed to form on the wing leading edge of the space shuttle after about 10-15 flights. In this report we expand upon previous observations by Christensen (1) that these pinholes often form along cracks and are associated with a locally zinc-rich area. The zinc appears to come from weathering and peeling paint on the launch structure. Three types of experimental examinations are performed to understand this issue further: (A) Detailed microstructural examination of actual shuttle pinholes (B) Mass spectrometric studies of coupons containing, actual shuttle pinholes and (C) Laboratory furnace studies of ZnO/SiC reactions and ZnO/SiC protected carbon/carbon reaction. On basis of these observations we present a detailed mechanism of pinhole formation due to formation of a corrosive ZnO-Na-2-O-SiO2 ternary glass, which flows into existing cracks and enlarges them.

Electron Microscopy Structural Insights into CPAP Oligomeric Behavior: A Plausible Assembly Process of a Supramolecular Scaffold of the Centrosome

PubMed Central

Alvarez-Cabrera, Ana L.; Delgado, Sandra; Gil-Carton, David; Mortuza, Gulnahar B.; Montoya, Guillermo; Sorzano, Carlos O. S.; Tang, Tang K.; Carazo, Jose M.

2017-01-01

Centrosomal P4.1-associated protein (CPAP) is a cell cycle regulated protein fundamental for centrosome assembly and centriole elongation. In humans, the region between residues 897–1338 of CPAP mediates interactions with other proteins and includes a homodimerization domain. CPAP mutations cause primary autosomal recessive microcephaly and Seckel syndrome. Despite of the biological/clinical relevance of CPAP, its mechanistic behavior remains unclear and its C-terminus (the G-box/TCP domain) is the only part whose structure has been solved. This situation is perhaps due in part to the challenges that represent obtaining the protein in a soluble, homogeneous state for structural studies. Our work constitutes a systematic structural analysis on multiple oligomers of HsCPAP897−1338, using single-particle electron microscopy (EM) of negatively stained (NS) samples. Based on image classification into clearly different regular 3D maps (putatively corresponding to dimers and tetramers) and direct observation of individual images representing other complexes of HsCPAP897−1338 (i.e., putative flexible monomers and higher-order multimers), we report a dynamic oligomeric behavior of this protein, where different homo-oligomers coexist in variable proportions. We propose that dimerization of the putative homodimer forms a putative tetramer which could be the structural unit for the scaffold that either tethers the pericentriolar material to centrioles or promotes procentriole elongation. A coarse fitting of atomic models into the NS 3D maps at resolutions around 20 Å is performed only to complement our experimental data, allowing us to hypothesize on the oligomeric composition of the different complexes. In this way, the current EM work represents an initial step toward the structural characterization of different oligomers of CPAP, suggesting further insights to understand how this protein works, contributing to the elucidation of control mechanisms for centriole

Space Shuttle Projects

NASA Image and Video Library

1984-04-24

The official mission insignia for the 41-D Space Shuttle flight features the Discovery - NASA's third orbital vehicle - as it makes its maiden voyage. The ghost ship represents the orbiter's namesakes which have figured prominently in the history of exploration. The Space Shuttle Discovery heads for new horizons to extend that proud tradition. Surnames for the crewmembers of NASA's eleventh Space Shuttle mission encircle the red, white, and blue scene.

Space Shuttle Projects

NASA Image and Video Library

1977-02-01

This photograph shows an inside view of a liquid hydrogen tank for the Space Shuttle external tank (ET) Main Propulsion Test Article (MPTA). The ET provides liquid hydrogen and liquid oxygen to the Shuttle's three main engines during the first 8.5 minutes of flight. At 154-feet long and more than 27-feet in diameter, the ET is the largest component of the Space Shuttle, the structural backbone of the entire Shuttle system, and is the only part of the vehicle that is not reusable. The ET is manufactured at the Michoud Assembly Facility near New Orleans, Louisiana, by the Martin Marietta Corporation under management of the Marshall Space Flight Center.

Space Shuttle Projects

NASA Image and Video Library

1978-05-01

This photograph shows a liquid oxygen tank for the Shuttle External Tank (ET) during a hydroelastic modal survey test at the Marshall Space Flight Center. The ET provides liquid hydrogen and liquid oxygen to the Shuttle's three main engines during the first 8.5 minutes of flight. At 154-feet long and more than 27-feet in diameter, the ET is the largest component of the Space Shuttle, the structural backbone of the entire Shuttle system, and is the only part of the vehicle that is not reusable. The ET is manufactured at the Michoud Assembly Facility near New Orleans, Louisiana, by the Martin Marietta Corporation under management of the Marshall Space Flight Center.

Shuttle interaction study extension

NASA Technical Reports Server (NTRS)

1981-01-01

The implications of using the Shuttle with the SOC were analyzed, including constraints that the Shuttle places upon the SOC design. All the considerations involved in the use of the shuttle as a part of the SOC concept were identified.

Loss of Ranbp2 in motoneurons causes disruption of nucleocytoplasmic and chemokine signaling, proteostasis of hnRNPH3 and Mmp28, and development of amyotrophic lateral sclerosis-like syndromes.

PubMed

Cho, Kyoung-In; Yoon, Dosuk; Qiu, Sunny; Danziger, Zachary; Grill, Warren M; Wetsel, William C; Ferreira, Paulo A

2017-05-01

The pathogenic drivers of sporadic and familial motor neuron disease (MND), such amyotrophic lateral sclerosis (ALS), are unknown. MND impairs the Ran GTPase cycle, which controls nucleocytoplasmic transport, ribostasis and proteostasis; however, cause-effect mechanisms of Ran GTPase modulators in motoneuron pathobiology have remained elusive. The cytosolic and peripheral nucleoporin Ranbp2 is a crucial regulator of the Ran GTPase cycle and of the proteostasis of neurological disease-prone substrates, but the roles of Ranbp2 in motoneuron biology and disease remain unknown. This study shows that conditional ablation of Ranbp2 in mouse Thy1 motoneurons causes ALS syndromes with hypoactivity followed by hindlimb paralysis, respiratory distress and, ultimately, death. These phenotypes are accompanied by: a decline in the nerve conduction velocity, free fatty acids and phophatidylcholine of the sciatic nerve; a reduction in the g-ratios of sciatic and phrenic nerves; and hypertrophy of motoneurons. Furthermore, Ranbp2 loss disrupts the nucleocytoplasmic partitioning of the import and export nuclear receptors importin β and exportin 1, respectively, Ran GTPase and histone deacetylase 4. Whole-transcriptome, proteomic and cellular analyses uncovered that the chemokine receptor Cxcr4, its antagonizing ligands Cxcl12 and Cxcl14, and effector, latent and activated Stat3 all undergo early autocrine and proteostatic deregulation, and intracellular sequestration and aggregation as a result of Ranbp2 loss in motoneurons. These effects were accompanied by paracrine and autocrine neuroglial deregulation of hnRNPH3 proteostasis in sciatic nerve and motoneurons, respectively, and post-transcriptional downregulation of metalloproteinase 28 in the sciatic nerve. Mechanistically, our results demonstrate that Ranbp2 controls nucleocytoplasmic, chemokine and metalloproteinase 28 signaling, and proteostasis of substrates that are crucial to motoneuronal homeostasis and whose impairments

7. YOSEMITE VALLEY SHUTTLE BUS AT SENTINEL BRIDGE SHUTTLE BUS ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

7. YOSEMITE VALLEY SHUTTLE BUS AT SENTINEL BRIDGE SHUTTLE BUS AND PARKING LOT AREA. LOOKING WNW. GIS: N-37 40 36.2 / W-119 44 45.0 - Yosemite National Park Roads & Bridges, Yosemite Village, Mariposa County, CA

Monitoring Shuttle Burns and Rocket Launches with GPS

NASA Astrophysics Data System (ADS)

Coster, A. J.; Bhatt, A.; O'Hanlon, B.; Rideout, W.

2009-12-01

We report on different GPS analysis techniques that can be used to examine the effects of rocket exhaust on the upper atmosphere. GPS observations of artificially produced electron density holes created by chemical releases from Space Shuttle Orbital Maneuvering System (OMS) engine burns will be discussed. The percentage drop in total electron content (TEC) and the temporal and spatial scales observed in the electron density hole for different Shuttle burn experiments will be compared. We will also report on observations of TEC depletions associated with Titan rocket launches on 8 April 2003 and on 19 October 2005. Finally we will discuss the use of GPS measurements of precipitable water vapor from time periods before, during, and after Shuttle burns.

Space Shuttle Projects

NASA Image and Video Library

1977-03-01

This photograph shows the liquid hydrogen tank and liquid oxygen tank for the Space Shuttle external tank (ET) being assembled in the weld assembly area of the Michoud Assembly Facility (MAF). The ET provides liquid hydrogen and liquid oxygen to the Shuttle's three main engines during the first eight 8.5 minutes of flight. At 154-feet long and more than 27-feet in diameter, the ET is the largest component of the Space Shuttle, the structural backbone of the entire Shuttle system, and the only part of the vehicle that is not reusable. The ET is manufactured at the Michoud Assembly Facility near New Orleans, Louisiana, by the Martin Marietta Corporation under management of the Marshall Space Flight Center.

Space Shuttle Discovery Launch

NASA Image and Video Library

2008-05-31

NASA Shuttle Launch Director Michael Leinbach, left, STS-124 Assistant Launch Director Ed Mango, center, and Flow Director for Space Shuttle Discovery Stephanie Stilson clap in the the Launch Control Center after the main engine cut off and successful launch of the Space Shuttle Discovery (STS-124) Saturday, May 31, 2008, at the Kennedy Space Center in Cape Canaveral, Fla. The Shuttle lifted off from launch pad 39A at 5:02 p.m. EDT. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Debris Transport

NASA Technical Reports Server (NTRS)

Gomez, Reynaldo J., III

2010-01-01

This slide presentation reviews the assessment of debris damage to the Space Shuttle, and the use of computation to assist in the space shuttle applications. The presentation reviews the sources of debris, a mechanism for determining the probability of damaging debris impacting the shuttle, tools used, eliminating potential damaging debris sources, the use of computation to assess while inflight damage, and a chart showing the applications that have been used on increasingly powerful computers simulate the shuttle and the debris transport.

History of Space Shuttle Rendezvous

NASA Technical Reports Server (NTRS)

Goodman, John L.

2011-01-01

This technical history is intended to provide a technical audience with an introduction to the rendezvous and proximity operations history of the Space Shuttle Program. It details the programmatic constraints and technical challenges encountered during shuttle development in the 1970s and over thirty years of shuttle missions. An overview of rendezvous and proximity operations on many shuttle missions is provided, as well as how some shuttle rendezvous and proximity operations systems and flight techniques evolved to meet new programmatic objectives. This revised edition provides additional information on Mercury, Gemini, Apollo, Skylab, and Apollo/Soyuz. Some chapters on the Space Shuttle have been updated and expanded. Four special focus chapters have been added to provide more detailed information on shuttle rendezvous. A chapter on the STS-39 mission of April/May 1991 describes the most complex deploy/retrieve mission flown by the shuttle. Another chapter focuses on the Hubble Space Telescope servicing missions. A third chapter gives the reader a detailed look at the February 2010 STS-130 mission to the International Space Station. The fourth chapter answers the question why rendezvous was not completely automated on the Gemini, Apollo, and Space Shuttle vehicles.

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (center) is given a tour of a solid rocket booster (SRB) retrieval ship by United Space Alliance (USA) employee Joe Chaput (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

NASA Image and Video Library

2003-12-19

KENNEDY SPACE CENTER, FLA. -- NASA Deputy Associate Administrator for Space Station and Shuttle Programs Michael Kostelnik (center) is given a tour of a solid rocket booster (SRB) retrieval ship by United Space Alliance (USA) employee Joe Chaput (right). NASA and USA Space Shuttle program management are participating in a leadership workday. The day is intended to provide management with an in-depth, hands-on look at Shuttle processing activities at KSC.

Space Shuttle Projects

NASA Image and Video Library

1994-09-13

Designed by the mission crew members, the STS-66 emblem depicts the Space Shuttle Atlantis launching into Earth orbit to study global environmental change. The payload for the Atmospheric Laboratory for Applications and Science (ATLAS-3) and complementary experiments were part of a continuing study of the atmosphere and the Sun's influence on it. The Space Shuttle is trailed by gold plumes representing the astronaut symbol and is superimposed over Earth, much of which is visible from the flight's high inclination orbit. Sensitive instruments aboard the ATLAS pallet in the Shuttle payload bay and on the free-flying Cryogenic Infrared Spectrometers and Telescopes for the Atmospheric-Shuttle Pallet Satellite (CHRISTA-SPAS) that gazed down on Earth and toward the Sun, are illustrated by the stylized sunrise and visible spectrum.

STS-124 Space Shuttle Discovery Landing

NASA Image and Video Library

2008-06-14

NASA Deputy Shuttle Program Manager LeRoy Cain points out a portion of the space shuttle Discovery to NASA Associate Administrator for Space Operations Bill Gerstenmaier, left, during a walk around shortly after Discovery touched down at 11:15 a.m., Saturday, June 14, 2008, at the Kennedy Space Center in Cape Canaveral, Florida. During the 14-day STS-124 mission Discovery's crew installed the Japan Aerospace Exploration Agency's large Kibo laboratory and its remote manipulator system leaving a larger space station and one with increased science capabilities. Discovery also brought home NASA astronaut Garrett Reisman after his 3 month mission onboard the International Space Station. Photo Credit: (NASA/Bill Ingalls)

53BP1 and USP28 mediate p53-dependent cell cycle arrest in response to centrosome loss and prolonged mitosis.

PubMed

Fong, Chii Shyang; Mazo, Gregory; Das, Tuhin; Goodman, Joshua; Kim, Minhee; O'Rourke, Brian P; Izquierdo, Denisse; Tsou, Meng-Fu Bryan

2016-07-02

Mitosis occurs efficiently, but when it is disturbed or delayed, p53-dependent cell death or senescence is often triggered after mitotic exit. To characterize this process, we conducted CRISPR-mediated loss-of-function screens using a cell-based assay in which mitosis is consistently disturbed by centrosome loss. We identified 53BP1 and USP28 as essential components acting upstream of p53, evoking p21-dependent cell cycle arrest in response not only to centrosome loss, but also to other distinct defects causing prolonged mitosis. Intriguingly, 53BP1 mediates p53 activation independently of its DNA repair activity, but requiring its interacting protein USP28 that can directly deubiquitinate p53 in vitro and ectopically stabilize p53 in vivo. Moreover, 53BP1 can transduce prolonged mitosis to cell cycle arrest independently of the spindle assembly checkpoint (SAC), suggesting that while SAC protects mitotic accuracy by slowing down mitosis, 53BP1 and USP28 function in parallel to select against disturbed or delayed mitosis, promoting mitotic efficiency.

Shuttle communications design study

NASA Technical Reports Server (NTRS)

Cartier, D. E.

1975-01-01

The design and development of a space shuttle communication system are discussed. The subjects considered include the following: (1) Ku-band satellite relay to shuttle, (2) phased arrays, (3) PN acquisition, (4) quadriplexing of direct link ranging and telemetry, (5) communications blackout on launch and reentry, (6) acquisition after blackout on reentry, (7) wideband communications interface with the Ku-Band rendezvous radar, (8) aeroflight capabilities of the space shuttle, (9) a triple multiplexing scheme equivalent to interplex, and (10) a study of staggered quadriphase for use on the space shuttle.

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

The space shuttle Endeavour is seen as it traverses through Inglewood, Calif. on Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Project

NASA Image and Video Library

1981-01-01

A Space Shuttle Main Engine undergoes test-firing at the National Space Technology Laboratories (now the Sternis Space Center) in Mississippi. The Marshall Space Flight Center had management responsibility of Space Shuttle propulsion elements, including the Main Engines.

Space Shuttle Program

NASA Image and Video Library

2012-09-12

Ronnie Rigney (r), chief of the Propulsion Test Office in the Project Directorate at Stennis Space Center, stands with agency colleagues to receive the prestigious American Institute of Aeronautics and Astronautics George M. Low Space Transportation Award on Sept. 12. Rigney accepted the award on behalf of the NASA and contractor team at Stennis for their support of the Space Shuttle Program that ended last summer. From 1975 to 2009, Stennis Space Center tested every main engine used to power 135 space shuttle missions. Stennis continued to provide flight support services through the end of the Space Shuttle Program in July 2011. The center also supported transition and retirement of shuttle hardware and assets through September 2012. The 2012 award was presented to the space shuttle team 'for excellence in the conception, development, test, operation and retirement of the world's first and only reusable space transportation system.' Joining Rigney for the award ceremony at the 2012 AIAA Conference in Pasadena, Calif., were: (l to r) Allison Zuniga, NASA Headquarters; Michael Griffin, former NASA administrator; Don Noah, Johnson Space Center in Houston; Steve Cash, Marshall Space Flight Center in Huntsville, Ala.; and Pete Nickolenko, Kennedy Space Center in Florida.

Space Shuttle Projects

NASA Image and Video Library

2004-04-15

The Apollo program demonstrated that men could travel into space, perform useful tasks there, and return safely to Earth. But space had to be more accessible. This led to the development of the Space Shuttle. The Shuttle's major components are the orbiter spacecraft; the three main engines, with a combined thrust of more than 1.2 million pounds; the huge external tank (ET) that feeds the liquid hydrogen fuel and liquid oxygen oxidizer to the three main engines; and the two solid rocket boosters (SRBs), with their combined thrust of some 5.8 million pounds, that provide most of the power for the first two minutes of flight. Crucially involved with the Space Shuttle program virtually from its inception, the Marshall Space Flight Center (MSFC) played a leading role in the design, development, testing, and fabrication of many major Shuttle propulsion components.

Space Shuttle Drawing

NASA Technical Reports Server (NTRS)

2004-01-01

The Apollo program demonstrated that men could travel into space, perform useful tasks there, and return safely to Earth. But space had to be more accessible. This led to the development of the Space Shuttle. The Shuttle's major components are the orbiter spacecraft; the three main engines, with a combined thrust of more than 1.2 million pounds; the huge external tank (ET) that feeds the liquid hydrogen fuel and liquid oxygen oxidizer to the three main engines; and the two solid rocket boosters (SRBs), with their combined thrust of some 5.8 million pounds, that provide most of the power for the first two minutes of flight. Crucially involved with the Space Shuttle program virtually from its inception, the Marshall Space Flight Center (MSFC) played a leading role in the design, development, testing, and fabrication of many major Shuttle propulsion components.

NASA Facts, Space Shuttle.

ERIC Educational Resources Information Center

National Aeronautics and Space Administration, Washington, DC. Educational Programs Div.

This newsletter from the National Aeronautics and Space Administration (NASA) contains a description of the purposes and potentials of the Space Shuttle craft. The illustrated document explains some of the uses for which the shuttle is designed; how the shuttle will be launched from earth, carry out its mission, and land again on earth; and what a…

Space Shuttle Project

NASA Image and Video Library

1972-03-07

This early chart conceptualizes the use of two parallel Solid Rocket Motor Boosters in conjunction with three main engines to launch the proposed Space Shuttle to orbit. At approximately twenty-five miles altitude, the boosters would detach from the Orbiter and parachute back to Earth where they would be recovered and refurbished for future use. The Shuttle was designed as NASA's first reusable space vehicle, launching vertically like a spacecraft and landing on runways like conventional aircraft. Marshall Space Flight Center had management responsibility for the Shuttle's propulsion elements, including the Solid Rocket Boosters.

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

The space shuttle Enterprise is lowered onto a transport vehicle after being demated from the NASA 747 Shuttle Carrier Aircraft (SCA) at John F. Kennedy (JFK) International Airport in Jamica, New York, Sunday, May 13, 2012. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to the Intrepid Sea, Air & Space Museum in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

The space shuttle Enterprise hangs from a sling after being demated from the NASA 747 Shuttle Carrier Aircraft (SCA) at John F. Kennedy (JFK) International Airport in Jamica, New York, Sunday, May 13, 2012. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to the Intrepid Sea, Air & Space Museum in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

A yellow sling is lowered onto space shuttle Enterprise, which sits atop NASA's 747 Shuttle Carrier Aircraft (SCA) prior to it being demated a few hours later at John F. Kennedy (JFK) International Airport in New York, Saturday, May 12, 2012. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to Intrepid in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

Spectators watch space shuttle Endeavour as it passes by on its way to its new home at the California Science Center in Los Angeles, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

The space shuttle Endeavour is seen as it traverses through the streest of Los Angeles on its way to its new home at the California Science Center, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

A spectator photographs the space shuttle Endeavour as it passes by on its way to its new home at the California Science Center in Los Angeles, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

A spectator is seen photographing the space shuttle Endeavour as it is moved to its new home at the California Science Center in Los Angeles, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC’s Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Bill Ingalls)

Post-Shuttle EVA Operations on ISS

NASA Technical Reports Server (NTRS)

West, William; Witt, Vincent; Chullen, Cinda

2010-01-01

The expected retirement of the NASA Space Transportation System (also known as the Space Shuttle ) by 2011 will pose a significant challenge to Extra-Vehicular Activities (EVA) on-board the International Space Station (ISS). The EVA hardware currently used to assemble and maintain the ISS was designed assuming that it would be returned to Earth on the Space Shuttle for refurbishment, or if necessary for failure investigation. With the retirement of the Space Shuttle, a new concept of operations was developed to enable EVA hardware (Extra-vehicular Mobility Unit (EMU), Airlock Systems, EVA tools, and associated support hardware and consumables) to perform ISS EVAs until 2015, and possibly beyond to 2020. Shortly after the decision to retire the Space Shuttle was announced, the EVA 2010 Project was jointly initiated by NASA and the One EVA contractor team. The challenges addressed were to extend the operating life and certification of EVA hardware, to secure the capability to launch EVA hardware safely on alternate launch vehicles, to protect for EMU hardware operability on-orbit, and to determine the source of high water purity to support recharge of PLSSs (no longer available via Shuttle). EVA 2010 Project includes the following tasks: the development of a launch fixture that would allow the EMU Portable Life Support System (PLSS) to be launched on-board alternate vehicles; extension of the EMU hardware maintenance interval from 3 years (current certification) to a minimum of 6 years (to extend to 2015); testing of recycled ISS Water Processor Assembly (WPA) water for use in the EMU cooling system in lieu of water resupplied by International Partner (IP) vehicles; development of techniques to remove & replace critical components in the PLSS on-orbit (not routine); extension of on-orbit certification of EVA tools; and development of an EVA hardware logistical plan to support the ISS without the Space Shuttle. Assumptions for the EVA 2010 Project included no more

Space Shuttle Project

NASA Image and Video Library

1978-10-04

The Shuttle Orbiter Enterprise inside of Marshall Space Flight Center's Dynamic Test Stand for Mated Vertical Ground Vibration tests (MVGVT). The tests marked the first time ever that the entire shuttle complement including Orbiter, external tank, and solid rocket boosters were vertically mated.

Orthostatic Intolerance After ISS and Space Shuttle Missions.

PubMed

Lee, Stuart M C; Feiveson, Alan H; Stein, Sydney; Stenger, Michael B; Platts, Steven H

2015-12-01

Cardiovascular deconditioning apparently progresses with flight duration, resulting in a greater incidence of orthostatic intolerance following long-duration missions. Therefore, we anticipated that the proportion of astronauts who could not complete an orthostatic tilt test (OTT) would be higher on landing day and the number of days to recover greater after International Space Station (ISS) than after Space Shuttle missions. There were 20 ISS and 65 Shuttle astronauts who participated in 10-min 80° head-up tilt tests 10 d before launch, on landing day (R+0), and 3 d after landing (R+3). Fisher's Exact Test was used to compare the ability of ISS and Shuttle astronauts to complete the OTT. Cox regression was used to identify cardiovascular parameters associated with OTT completion and mixed model analysis was used to compare the change and recovery rates between groups. The proportion of astronauts who completed the OTT on R+0 (2 of 6) was less in ISS than in Shuttle astronauts (52 of 65). On R+3, 13 of 15 and 19 of 19 of the ISS and Shuttle astronauts, respectively, completed the OTT. An index comprised of stroke volume and diastolic blood pressure provided a good prediction of OTT completion and was altered by spaceflight similarly for both astronaut groups, but recovery was slower in ISS than in Shuttle astronauts. The proportion of ISS astronauts who could not complete the OTT on R+0 was greater and the recovery rate slower after ISS compared to Shuttle missions. Thus, mission planners and crew surgeons should anticipate the need to tailor scheduled activities and level of medical support to accommodate protracted recovery after long-duration microgravity exposures.

HIV-1 Nef-associated Factor 1 Enhances Viral Production by Interacting with CRM1 to Promote Nuclear Export of Unspliced HIV-1 gag mRNA.

PubMed

Ren, Xiao-Xin; Wang, Hai-Bo; Li, Chuan; Jiang, Jin-Feng; Xiong, Si-Dong; Jin, Xia; Wu, Li; Wang, Jian-Hua

2016-02-26

HIV-1 depends on host-cell-encoded factors to complete its life cycle. A comprehensive understanding of how HIV-1 manipulates host machineries during viral infection can facilitate the identification of host targets for antiviral drugs or gene therapy. The cellular protein Naf1 (HIV-1 Nef-associated factor 1) is a CRM1-dependent nucleo-cytoplasmic shuttling protein, and has been identified to regulate multiple receptor-mediated signal pathways in inflammation. The cytoplasm-located Naf1 can inhibit NF-κB activation through binding to A20, and the loss of Naf1 controlled NF-κB activation is associated with multiple autoimmune diseases. However, the effect of Naf1 on HIV-1 mRNA expression has not been characterized. In this study we found that the nucleus-located Naf1 could promote nuclear export of unspliced HIV-1 gag mRNA. We demonstrated that the association between Naf1 and CRM1 was required for this function as the inhibition or knockdown of CRM1 expression significantly impaired Naf1-promoted HIV-1 production. The mutation of Naf1 nuclear export signals (NESs) that account for CRM1 recruitment for nuclear export decreased Naf1 function. Additionally, the mutation of the nuclear localization signal (NLS) of Naf1 diminished its ability to promote HIV-1 production, demonstrating that the shuttling property of Naf1 is required for this function. Our results reveal a novel role of Naf1 in enhancing HIV-1 production, and provide a potential therapeutic target for controlling HIV-1 infection. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

STS-52 Space Shuttle mission report

NASA Technical Reports Server (NTRS)

Fricke, Robert W., Jr.

1992-01-01

The STS-52 Space Shuttle Program Mission Report provides a summary of the Orbiter, External Tank (ET), Solid Rocket Booster/Redesigned Solid Rocket Motor (SRB/RSRM), and the Space Shuttle main engine (SSME) subsystem performance during the fifty-first flight of the Space Shuttle Program, and the thirteenth flight of the Orbiter vehicle Columbia (OV-102). In addition to the Orbiter, the flight vehicle consisted of the following: an ET (designated as ET-55/LWT-48); three SSME's, which were serial numbers 2030, 2015, and 2034 in positions 1, 2, and 3, respectively; and two SRB's, which were designated BI-054. The lightweight RSRM's that were installed in each SRB were designated 360L027A for the left SRB and 360Q027B for the right SRB. The primary objectives of this flight were to successfully deploy the Laser Geodynamic Satellite (LAGEOS-2) and to perform operations of the United States Microgravity Payload-1 (USMP-1). The secondary objectives of this flight were to perform the operations of the Attitude Sensor Package (ASP), the Canadian Experiments-2 (CANEX-2), the Crystals by Vapor Transport Experiment (CVTE), the Heat Pipe Performance Experiment (HPP), the Commercial Materials Dispersion Apparatus Instrumentation Technology Associates Experiments (CMIX), the Physiological System Experiment (PSE), the Commercial Protein Crystal Growth (CPCG-Block 2), the Shuttle Plume Impingement Experiment (SPIE), and the Tank Pressure Control Experiment (TPCE) payloads.

STS-52 Space Shuttle mission report

NASA Astrophysics Data System (ADS)

Fricke, Robert W., Jr.

1992-12-01

The STS-52 Space Shuttle Program Mission Report provides a summary of the Orbiter, External Tank (ET), Solid Rocket Booster/Redesigned Solid Rocket Motor (SRB/RSRM), and the Space Shuttle main engine (SSME) subsystem performance during the fifty-first flight of the Space Shuttle Program, and the thirteenth flight of the Orbiter vehicle Columbia (OV-102). In addition to the Orbiter, the flight vehicle consisted of the following: an ET (designated as ET-55/LWT-48); three SSME's, which were serial numbers 2030, 2015, and 2034 in positions 1, 2, and 3, respectively; and two SRB's, which were designated BI-054. The lightweight RSRM's that were installed in each SRB were designated 360L027A for the left SRB and 360Q027B for the right SRB. The primary objectives of this flight were to successfully deploy the Laser Geodynamic Satellite (LAGEOS-2) and to perform operations of the United States Microgravity Payload-1 (USMP-1). The secondary objectives of this flight were to perform the operations of the Attitude Sensor Package (ASP), the Canadian Experiments-2 (CANEX-2), the Crystals by Vapor Transport Experiment (CVTE), the Heat Pipe Performance Experiment (HPP), the Commercial Materials Dispersion Apparatus Instrumentation Technology Associates Experiments (CMIX), the Physiological System Experiment (PSE), the Commercial Protein Crystal Growth (CPCG-Block 2), the Shuttle Plume Impingement Experiment (SPIE), and the Tank Pressure Control Experiment (TPCE) payloads.

Space operations center: Shuttle interaction study extension, executive summary

NASA Technical Reports Server (NTRS)

1982-01-01

The Space Operations Center (SOC) is conceived as a permanent facility in low Earth orbit incorporating capabilities for space systems construction; space vehicle assembly, launching, recovery and servicing; and the servicing of co-orbiting satellites. The Shuttle Transportation System is an integral element of the SOC concept. It will transport the various elements of the SOC into space and support the assembly operation. Subsequently, it will regularly service the SOC with crew rotations, crew supplies, construction materials, construction equipment and components, space vehicle elements, and propellants and spare parts. The implications to the SOC as a consequence of the Shuttle supporting operations are analyzed. Programmatic influences associated with propellant deliveries, spacecraft servicing, and total shuttle flight operations are addressed.

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

NASA's 747 Shuttle Carrier Aircraft (SCA), with space shuttle Enterprise latched on its back, is towed from the hangar at John F. Kennedy (JFK) International Airport in New York late in the night on Saturday, May 12, 2012. Early Sunday morning, Enterprise was removed from the SCA. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to Intrepid in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

The driver of the Over Land Transporter is seen as he maneuvers the space shuttle Endeavour on the streets of Los Angeles as it heads to its new home at the California Science Center, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC’s Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

Spectators are seen as they watch space shuttle Endeavour as it passes by on its way to its new home at the California Science Center in Los Angeles, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

The driver of the Over Land Transporter (OLT) is seen as he maneuvers the space shuttle Endeavour on the streets of Los Angeles as it heads to its new home at the California Science Center, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

A spectator on the roof of a building photographs space shuttle Endeavour as it passes by on its way to its new home at the California Science Center in Los Angeles, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC’s Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Endeavour Move

NASA Image and Video Library

2012-10-12

The space shuttle Endeavour moves out of the Los Angeles International Airport and onto the streets of Los Angeles to make its way to its new home at the California Science Center, Friday, Oct. 12, 2012. Endeavour, built as a replacement for space shuttle Challenger, completed 25 missions, spent 299 days in orbit, and orbited Earth 4,671 times while traveling 122,883,151 miles. Beginning Oct. 30, the shuttle will be on display in the CSC's Samuel Oschin Space Shuttle Endeavour Display Pavilion, embarking on its new mission to commemorate past achievements in space and educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Space Shuttle Hot Cabin Emergency Responses

NASA Technical Reports Server (NTRS)

Stepaniak, P.; Effenhauser, R. K.; McCluskey, R.; Gillis, D. B.; Hamilton, D.; Kuznetz, L. H.

2005-01-01

Methods: Human thermal tolerance, countermeasures, and thermal model data were reviewed and compared to existing shuttle ECS failure temperature and humidity profiles for each failure mode. Increases in core temperature associated with cognitive impairment was identified, as was metabolic heat generation of crewmembers, temperature monitoring, and communication capabilities after partial power-down and other limiting factors. Orbiter landing strategies and a hydration and salt replacement protocol were developed to put wheels on deck in each failure mode prior to development of significant cognitive impairment or collapse of crewmembers. Thermal tradeoffs for use of the Advanced Crew Escape Suit (ACES), Liquid Cooling Garment, integrated G-suit and Quick Don Mask were examined. candidate solutions involved trade-offs or conflicts with cabin oxygen partial pressure limits, system power-downs to limit heat generation, risks of alternate and emergency landing sites or compromise of Mode V-VIII scenarios. Results: Rehydration and minimized cabin workloads are required in all failure modes. Temperature/humidity profiles increase rapidly in two failure modes, and deorbit is recommended without the ACES, ICU and g-suit. This latter configuration limits several shuttle approach and landing escape modes and requires communication modifications. Additional data requirements were identified and engineering simulations were recommended to develop more current shuttle temperature and humidity profiles. Discussion: After failure of the shuttle ECS, there is insufficient cooling capacity of the ACES to protect crewmembers from rising cabin temperature and humidity. The LCG is inadequate for cabin temperatures above 76 F. Current shuttle future life policy makes it unlikely that major engineering upgrades necessary to address this problem will occur.

Space Shuttle Projects

NASA Image and Video Library

1978-03-01

A liquid hydrogen tank of the Shuttle's external tank (ET) is installed into the S-1C Test Stand for a structural test at the Marshall Space Flight Center. At 154-feet long and more than 27-feet in diameter, the ET is the largest component of the Space Shuttle, the structural backbone of the entire Shuttle system, and is the only part of the vehicle that is not reusable. The ET is manufactured at the Michoud Assembly Facility near New Orleans, Louisiana, by the Martin Marietta Corporation under management of the Marshall Space Flight Center.

Assessment Regarding Impact of Atmospheric Conditions on Space Shuttle Launch Delays

NASA Technical Reports Server (NTRS)

Johnson D. L.; Pearson, S. D.; Vaughan, W. W.; Batts, G. W.

1998-01-01

The atmospheric environment definition has played a key role in the development and operation of the NASA Space Shuttle as it has in other NASA Space Vehicle Programs. The objective of any definition of natural environment design requirements for a space vehicle development is to insure that the vehicle will perform safely and in a timely manner relative to the mission(s) for which the vehicle is being developed. The NASA Space Shuttle has enjoyed the longest tenure of any Space Vehicle from an operational standpoint. As such, it has provided a wealth of information on many engineering aspects of a Space Vehicle plus the influence of the atmosphere on operational endeavors. The atmospheric environment associated with the NASA Space Shuttle launches at the NASA Kennedy Space Center in Florida has been reviewed and studied over the entire NASA Space Shuttle flight history. The results of the analysis of atmospheric environment related launch delays relative to other sources of launch delays has been assessed. This paper will provide a summary of those conditions as well as mission analysis examples focused on atmospheric constraints for launch. Atmospheric conditions associated with NASA Space Shuttle launch delays will be presented to provide a reference as to the type conditions experienced which have mainly caused the delays.

Space Shuttle Strategic Planning Status

NASA Technical Reports Server (NTRS)

Henderson, Edward M.; Norbraten, Gordon L.

2006-01-01

The Space Shuttle Program is aggressively planning the Space Shuttle manifest for assembling the International Space Station and servicing the Hubble Space Telescope. Implementing this flight manifest while concurrently transitioning to the Exploration architecture creates formidable challenges; the most notable of which is retaining critical skills within the Shuttle Program workforce. The Program must define a strategy that will allow safe and efficient fly-out of the Shuttle, while smoothly transitioning Shuttle assets (both human and facility) to support early flight demonstrations required in the development of NASA s Crew Exploration Vehicle (CEV) and Crew and Cargo Launch Vehicles (CLV). The Program must accomplish all of this while maintaining the current level of resources. Therefore, it will be necessary to initiate major changes in operations and contracting. Overcoming these challenges will be essential for NASA to fly the Shuttle safely, accomplish the President s "Vision for Space Exploration," and ultimately meet the national goal of maintaining a robust space program. This paper will address the Space Shuttle Program s strategy and its current status in meeting these challenges.

Space Shuttle Strategic Planning Status

NASA Technical Reports Server (NTRS)

Norbraten, Gordon L.; Henderson, Edward M.

2007-01-01

The Space Shuttle Program is aggressively flying the Space Shuttle manifest for assembling the International Space Station and servicing the Hubble Space Telescope. Completing this flight manifest while concurrently transitioning to the Exploration architecture creates formidable challenges; the most notable of which is retaining critical skills within the Shuttle Program workforce. The Program must define a strategy that will allow safe and efficient fly-out of the Shuttle, while smoothly transitioning Shuttle assets (both human and facility) to support early flight demonstrations required in the development of NASA's Crew Exploration Vehicle (Orion) and Crew and Cargo Launch Vehicles (Ares I). The Program must accomplish all of this while maintaining the current level of resources. Therefore, it will be necessary to initiate major changes in operations and contracting. Overcoming these challenges will be essential for NASA to fly the Shuttle safely, accomplish the Vision for Space Exploration, and ultimately meet the national goal of maintaining a robust space program. This paper will address the Space Shuttle Program s strategy and its current status in meeting these challenges.

Space Shuttle Project

NASA Image and Video Library

1978-04-21

The Shuttle Orbiter Enterprise is lowered into the Dynamic Test Stand for Mated Vertical Ground Vibration tests (MVGVT) at the Marshall Space Flight Center. The tests marked the first time ever that the entire shuttle complement (including Orbiter, external tank, and solid rocket boosters) were mated vertically.

Shuttle Discovery Landing at Edwards

NASA Technical Reports Server (NTRS)

1989-01-01

The STS-29 Space Shuttle Discovery mission lands at NASA's then Ames-Dryden Flight Research Facility, Edwards AFB, California, early Saturday morning, 18 March 1989. Touchdown was at 6:35:49 a.m. PST and wheel stop was at 6:36:40 a.m. on runway 22. Controllers chose the concrete runway for the landing in order to make tests of braking and nosewheel steering. The STS-29 mission was very successful, completing the launch of a Tracking and Data Relay communications satellite, as well as a range of scientific experiments. Discovery's five-man crew was led by Commander Michael L. Coats, and included pilot John E. Blaha and mission specialists James P. Bagian, Robert C. Springer, and James F. Buchli. Space Shuttles are the main element of America's Space Transportation System and are used for space research and other space applications. The shuttles are the first vehicles capable of being launched into space and returning to Earth on a routine basis. Space Shuttles are used as orbiting laboratories in which scientists and mission specialists conduct a wide variety of scientific experiments. Crews aboard shuttles place satellites in orbit, rendezvous with satellites to carry out repair missions and return them to space, and retrieve satellites and return them to Earth for refurbishment and reuse. Space Shuttles are true aerospace vehicles. They leave Earth and its atmosphere under rocket power provided by three liquid-propellant main engines with two solid-propellant boosters attached plus an external liquid-fuel tank. After their orbital missions, they streak back through the atmosphere and land like airplanes. The returning shuttles, however, land like gliders, without power and on runways. Other rockets can place heavy payloads into orbit, but, they can only be used once. Space Shuttles are designed to be continually reused. When Space Shuttles are used to transport complete scientific laboratories into space, the laboratories remain inside the payload bay throughout

Space Shuttle Project

NASA Image and Video Library

1978-10-04

The Shuttle Orbiter Enterprise is being installed into liftoff configuration at Marshall Space Flight Center's Dynamic Test Stand for Mated Vertical Ground Vibration tests (MVGVT). The tests marked the first time ever that the entire shuttle complement (including Orbiter, external tank, and solid rocket boosters) were mated vertically.

Shuttle car loading system

NASA Technical Reports Server (NTRS)

Collins, E. R., Jr. (Inventor)

1985-01-01

A system is described for loading newly mined material such as coal, into a shuttle car, at a location near the mine face where there is only a limited height available for a loading system. The system includes a storage bin having several telescoping bin sections and a shuttle car having a bottom wall that can move under the bin. With the bin in an extended position and filled with coal the bin sections can be telescoped to allow the coal to drop out of the bin sections and into the shuttle car, to quickly load the car. The bin sections can then be extended, so they can be slowly filled with more while waiting another shuttle car.

Shuttle Wastewater Solution Characterization

NASA Technical Reports Server (NTRS)

Adam, Niklas; Pham, Chau

2011-01-01

During the 31st shuttle mission to the International Space Station, STS-129, there was a clogging event in the shuttle wastewater tank. A routine wastewater dump was performed during the mission and before the dump was completed, degraded flow was observed. In order to complete the wastewater dump, flow had to be rerouted around the dump filter. As a result, a basic chemical and microbial investigation was performed to understand the shuttle wastewater system and perform mitigation tasks to prevent another blockage. Testing continued on the remaining shuttle flights wastewater and wastewater tank cleaning solutions. The results of the analyses and the effect of the mitigation steps are detailed in this paper.

Materials and processes for shuttle engine, external tank, and solid rocket booster

NASA Technical Reports Server (NTRS)

Schwinghamer, R. J.

1977-01-01

The Shuttle flight system is composed of the Orbiter, an External Tank (ET) that contains the ascent propellant to be used by the Space Shuttle Main Engines (SSME), and two Solid Rocket Boosters (SRB). The ET is expended on each launch; the Orbiter and SRB's are reusable. It is the requirement for reuse which poses the exciting new materials and processes challenges in the development of the Space Shuttle. A brief description of the Space Shuttle and the mission profile is given. The Shuttle configuration is then described with emphasis on the SSME, ET, and SRB. The materials selection, tracking, and control system used to assure reliability and to minimize cost are described, and salient features and challenges in materials and processes associated with the SSME, ET, and SRB are subsequently discussed.

PROTEINS IN NUCLEOCYTOPLASMIC INTERACTIONS

PubMed Central

Prescott, David; Goldstein, Lester

1968-01-01

The behavior of nuclear proteins in Amoeba proteus was studied by tritiated amino acid labeling, nuclear transplantation, and cytoplasmic amputation. During prophase at least 77% (but probably over 95%) of the nuclear proteins is released to the cytoplasm. These same proteins return to the nucleus within the first 3 hr of interphase. When cytoplasm is amputated from an ameba in mitosis (shen the nuclear proteins are in the cytoplasm), the resultant daughter nuclei are depleted in the labeled nuclear proteins. The degree of depletion is less than proportional to the amount of cytoplasm removed because a portion of rapidly migrating protein (a nuclear protein that is normally shuttling between nucleus and cytoplasm and is thus also present in the cytoplasm) which would normally remain in the cytoplasm is taken up by the reconstituting daughter nuclei. Cytoplasmic fragments cut from mitotic cells are enriched in both major classes of nuclear proteins, i.e. rapidly migrating protein and slow turn-over protein. An interphase nucleus implanted into such an enucleated cell acquires from the cytoplasm essentially all of the excess nuclear proteins of both classes. The data indicate that there is a lack of binding sites in the cytoplasm for the rapidly migrating nuclear protein. The quantitative aspects of the distribution of rapidly migrating protein between the nucleus and the cytoplasm indicate that the distribution is governed primarily by factors within the nucleus. PMID:5677972

Motility and centrosomal organization during sea urchin and mouse fertilization

NASA Technical Reports Server (NTRS)

Schatten, Heide; Schatten, Gerald

1986-01-01

It is noted that microfilaments are essential for incorporation of sperm in sea urchins and for pronuclear apposition in mice. The ability of sea urchin sperm to fertilize eggs is lowered by latrunculin, giving evidence that acrosomal microfilaments are of importance to the process of fertilization. Due to the uncertainty regarding the presence of microfilaments in various mammalian sperm, it is interesting that latrunculin does not noticeably affect the ability of mouse sperm to fertilize oocytes. The movements of the sperm and egg nuclei at the time of sea urchin fertilization are dependent on microtubules arranged into a radial monastral array (the sperm aster). In the mouse egg, microtubule activity is also required during pronuclear apposition, but they are arranged by a number of egg cytoplasmic sites. Results of the investigations show that both microtubules and microfilaments are necessary for the successful completion of fertilization in both mice and sea urchins, but at different stages. Also, it is demonstrated that centrosomes are contributed by the sperm in the process of sea urchin fertilization, but in mammals they may be inherited maternally.

Solid rocket motors for the Space Shuttle booster.

NASA Technical Reports Server (NTRS)

Odom, J. B.

1972-01-01

The evolution of the space shuttle booster system is reviewed from its initial concepts based on liquid-propellant reusable boosters to the final selection of recoverable, solid-fuel rocket motors. The rationale associated with each of the several major decisions in the evolution process is discussed. It is shown that the external tank orbiter configuration emerging from the latest studies takes maximum advantage of the solid rocket motor development experience and promises to be the optimum configuration for fulfilling the paramount shuttle program requirements of minimum total development risk within acceptable costs.

The Space Shuttle

NASA Technical Reports Server (NTRS)

Moffitt, William L.

2003-01-01

As missions have become increasingly more challenging over the years, the most adaptable and capable element of space shuttle operations has proven time and again to be human beings. Human space flight provides unique aspects of observation. interaction and intervention that can reduce risk and improve mission success. No other launch vehicle - in development or in operation today - can match the space shuttle's human space flight capabilities. Preserving U.S. leadership in human space flight requires a strategy to meet those challenges. The ongoing development of next generation vehicles, along with upgrades to the space shuttle, is the most effective means for assuring our access to space.

TERSSE. Definition of the total earth resources system for the shuttle era. Volume 9: Earth resources shuttle applications

NASA Technical Reports Server (NTRS)

Alverado, U.

1975-01-01

The use of the space shuttle for the Earth Resources Program is discussed. Several problems with respect to payload selection, integration, and mission planning were studied. Each of four shuttle roles in the sortie mode were examined and projected into an integrated shuttle program. Several representative Earth Resources missions were designed which would use the shuttle sortie as a platform and collectively include the four shuttle roles. An integrated flight program based on these missions was then developed for the first two years of shuttle flights. A set of broad implications concerning the uses of the shuttle for Earth Resources studies resulted.

Solar flares, proton showers, and the Space Shuttle

NASA Technical Reports Server (NTRS)

Rust, D. M.

1982-01-01

Attention is given the hazards posed to Space Shuttle crews by energetic proton radiation from inherently unpredictable solar flares, such as that of April 10-13, 1981, which was experienced by the Space Shuttle Columbia. The most energetic protons from this flare reached the earth's atmosphere an hour after flare onset, and would have posed a potentially lethal threat to astronauts engaged in extravehicular activity in a polar or geosynchronous orbit rather than the low-latitude, low-altitude orbit of this mission. It is shown that proton-producing flares are associated with energization in shocks, many of which are driven by coronal mass ejections. Insights gained from the Solar Maximum Year programs allow reconsideration of proton shower forecasting, which will be essential in the prediction of the weather that Space Shuttle astronauts will encounter during extravehicular activities.

Analysis of nuclear export using photoactivatable GFP fusion proteins and interspecies heterokaryons.

PubMed

Nakrieko, Kerry-Ann; Ivanova, Iordanka A; Dagnino, Lina

2010-01-01

In this chapter, we review protocols for the analysis of nucleocytoplasmic shuttling of transcription factors and nuclear proteins, using two different approaches. The first involves the use of photoactivatable forms of the protein of interest by fusion to photoactivatable green fluorescent protein to follow its movement out of the nucleus by live-cell confocal microscopy. This methodology allows for the kinetic characterization of protein movements as well as measurement of steady-state levels. In a second procedure to assess the ability of a nuclear protein to move into and out of the nucleus, we describe the use of interspecies heterokaryon assays, which provide a measurement of steady-state distribution. These technologies are directly applicable to the analysis of nucleocytoplasmic movements not only of transcription factors, but also other nuclear proteins.

Space Shuttle Projects

NASA Image and Video Library

1985-04-01

In this photograph the SYNCOM IV-3, also known as LEASAT 3, satellite moves away from the Space Shuttle Orbiter Discovery. SYNCOM (Hughes Geosynchronous Communication Satellite) provides communication services from geosynchronous orbit, principally to the U.S. Government. The satellite was launched on April 12, 1985, aboard the Space Shuttle Orbiter Discovery.

The X11L/X11{beta}/MINT2 and X11L2/X11{gamma}/MINT3 scaffold proteins shuttle between the nucleus and cytoplasm

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sumioka, Akio; Saito, Yuhki; Sakuma, Megumi

2008-03-10

The X11/MINT family proteins are adaptor scaffolding proteins involved in formation of multiprotein complexes, and trafficking and metabolism of membrane proteins such as the beta-amyloid precursor protein. We found that a significant portion of X11L and X11L2 are recovered in nuclear fraction of mouse brain homogenates. EGFP-X11s were not detected in the nucleus of N2a neuroblastoma cells; however, administration of leptomycin B (LMB) induced substantial nuclear accumulation of EGFP-X11L and EGFP-X11L2, while EGFP-X11 showed little accumulation. Fluorescence loss in photobleaching (FLIP) analysis indicated that EGFP-X11L2 and EGFP-X11L are shuttled between the cytoplasm and nucleus, the former more effectively than themore » latter. We identified a nuclear export signal (NES) in the N-terminus of X11L2, mutation of which induces nuclear accumulation of EGFP-X11L2 in the absence of LMB. X11L2 fused to the Gal4 DNA binding domain (DBD) showed transcriptional activity, suggesting that X11L2 could function as a transcriptional activator if tethered near a promoter. Interestingly, attenuation of the nucleo-cytoplasmic shuttling of GAL4-DBD-X11L2 by mutating the NES or attaching the SV40 nuclear localization signal significantly decreased the apparent transcriptional activity. Our observations suggest that X11L2 functions in the nucleus by a mechanism distinct from conventional transactivators.« less

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

Space shuttle Enterprise is held aloft by a yellow sling and a set of cranes after it was removed from the top of NASA's 747 Shuttle Carrier Aircraft early Sunday morning at John F. Kennedy (JFK) International Airport in New York, Sunday, May 13, 2012 .The 747 was towed backwards so that Enterprise could be lowered. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to the Intrepid Sea, Air & Space Museum in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

NASA and United Space Alliance workers lower a yellow sling onto space shuttle Enterprise, which sits atop NASA's 747 Shuttle Carrier Aircraft (SCA) prior to it being demated a few hours later at John F. Kennedy (JFK) International Airport in New York, Saturday, May 12, 2012. Once the sling was firmly attached early Sunday morning, Enterprise was lifted from the SCA. The shuttle will be placed on a barge that will move by tugboat up the Hudson River to Intrepid in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

NASA space shuttle lightweight seat

NASA Technical Reports Server (NTRS)

Hansen, Chris; Jermstad, Wayne; Lewis, James; Colangelo, Todd

1996-01-01

The Space Shuttle Lightweight Seat-Mission Specialist (LWS-MS) is a crew seat for the mission specialists who fly aboard the Space Shuttle. The LWS-MS is a lightweight replacement for the mission specialist seats currently flown on the Shuttle. Using state-of-the-art analysis techniques, a team of NASA and Lockheed engineers from the Johnson Space Center (JSC) designed a seat that met the most stringent requirements demanded of the new seats by the Shuttle program, and reduced the weight of the seats by 52%.

Intersectin goes nuclear: secret life of an endocytic protein.

PubMed

Alvisi, Gualtiero; Paolini, Lucia; Contarini, Andrea; Zambarda, Chiara; Di Antonio, Veronica; Colosini, Antonella; Mercandelli, Nicole; Timmoneri, Martina; Palù, Giorgio; Caimi, Luigi; Ricotta, Doris; Radeghieri, Annalisa

2018-04-27

Intersectin 1-short (ITSN1-s) is a 1220 amino acid ubiquitously expressed scaffold protein presenting a multidomain structure that allows to spatiotemporally regulate the functional interaction of a plethora of proteins. Besides its well-established role in endocytosis, ITSN1-s is involved in the regulation of cell signaling and is implicated in tumorigenesis processes, although the signaling pathways involved are still poorly understood. Here, we identify ITSN1-s as a nucleocytoplasmic trafficking protein. We show that, by binding to importin (IMP)α, a small fraction of ITSN1-s localizes in the cell nucleus at the steady state, where it preferentially associates with the nuclear envelope and interacts with lamin A/C. However, upon pharmacological ablation of chromosome region maintenance 1 (CRM-1)-dependent nuclear export pathway, the protein accumulates into the nucleus, thus revealing its moonlighting nature. Analysis of deletion mutants revealed that the coiled coil (CC) and Src homology (SH3) regions play the major role in its nucleocytoplasmic shuttling. While no evidence of nuclear localization signal (NLS) was detected in the CC region, a functional bipartite NLS was identified within the SH3D region of ITSN1-s (RKKNPGGWWEGELQARGKKRQIGW-1127), capable of conferring energy-dependent nuclear accumulation to reporter proteins and whose mutational ablation affects nuclear import of the whole SH3 region. Thus, ITSN1-s is an endocytic protein, which shuttles between the nucleus and the cytoplasm in a CRM-1- and IMPα-dependent fashion. © 2018 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Shuttle - Mir Program Insignia

NASA Image and Video Library

1994-09-20

The rising sun signifies the dawn of a new era of human Spaceflight, the first phase of the United States/Russian space partnership, Shuttle-Mir. Mir is shown in its proposed final on orbit configuration. The Shuttle is shown in a generic tunnel/Spacehab configuration. The Shuttle/Mir combination, docked to acknowledge the union of the two space programs, orbits over an Earth devoid of any definable features or political borders to emphasize Earth as the home planet for all humanity. The individual stars near the Space Shuttle and the Russian Mir Space Station represent the previous individual accomplishments of Russia's space program and that of the United States. The binary star is a tribute to the previous United States-Russian joint human Spaceflight program, the Apollo-Soyuz Test Project (ASTP). The flags of the two nations are symbolized by flowing ribbons of the national colors interwoven in space to represent the two nations joint exploration of space. NASA SHUTTLE and PKA MNP are shown in the stylized logo fonts of the two agencies that are conducting this program.

Ergonomic assessment of airport shuttle driver tasks using an ergonomic analysis toolset.

PubMed

Çakıt, Erman

2018-06-01

This study aimed to (a) evaluate strength requirements and lower back stresses during lifting and baggage handling tasks with the 3D Static Strength Prediction Program (3DSSPP) and (b) provide additional analyses using rapid entire body assessment (REBA) and the NASA task load index (TLX) to assess the risks associated with the tasks. Four healthy female shuttle drivers of good health aged between 55 and 60 years were observed and interviewed in an effort to determine the tasks required of their occupations. The results indicated that lifting bags and placing them in a shuttle were high risk for injury and possible changes should be further investigated. The study concluded there was a potential for injury associated with baggage storing and retrieval tasks of a shuttle driver.

Space Shuttle Damper System for Ground Wind Load Tests

NASA Technical Reports Server (NTRS)

Robinson, G. D.; Holt, J. R.; Chang, C. S.

1973-01-01

An active damper system which was originally developed for a 5.5% Saturn IB/Skylab Ground Winds Model was modified and used for similar purposes in a Space Shuttle model. A second damper system which was originally used in a 3% Saturn V/Dry Workshop model was also modified and made compatible with the Space Shuttle model to serve as a back-up system. Included in this final report are descriptions of the modified damper systems and the associated control and instrumentation.

Shuttle interaction study extension

NASA Technical Reports Server (NTRS)

1982-01-01

The following areas of Space Shuttle technology were discussed: variable altitude strategy, spacecraft servicing, propellant storage, orbiter plume impingement, space based design, mating (docking and berthing), shuttle fleet utilization, and mission/traffic model.

Shuttle Risk Progression by Flight

NASA Technical Reports Server (NTRS)

Hamlin, Teri; Kahn, Joe; Thigpen, Eric; Zhu, Tony; Lo, Yohon

2011-01-01

Understanding the early mission risk and progression of risk as a vehicle gains insights through flight is important: . a) To the Shuttle Program to understand the impact of re-designs and operational changes on risk. . b) To new programs to understand reliability growth and first flight risk. . Estimation of Shuttle Risk Progression by flight: . a) Uses Shuttle Probabilistic Risk Assessment (SPRA) and current knowledge to calculate early vehicle risk. . b) Shows impact of major Shuttle upgrades. . c) Can be used to understand first flight risk for new programs.

Stennis certifies final shuttle engine

NASA Image and Video Library

2008-10-22

Steam blasts out of the A-2 Test Stand at Stennis Space Center on Oct. 22 as engineers begin a certification test on engine 2061, the last space shuttle main flight engine scheduled to be built. Since 1975, Stennis has tested every space shuttle main engine used in the program - about 50 engines in all. Those engines have powered more than 120 shuttle missions - and no mission has failed as a result of engine malfunction. For the remainder of 2008 and throughout 2009, Stennis will continue testing of various space shuttle main engine components.

Space Shuttle Project

NASA Image and Video Library

1978-04-21

This is an interior ground level view of the Shuttle Orbiter Enterprise being lowered for mating to External Tank (ET) inside Marshall Space Flight Center's Dynamic Test Stand for Mated Vertical Ground Vibration tests (MVGVT). The tests marked the first time ever that the entire shuttle complement (including Orbiter, external tank, and solid rocket boosters) were mated vertically.

Mustard Gas Surrogate, 2-Chloroethyl Ethylsulfide (2-CEES), Induces Centrosome Amplification and Aneuploidy in Human and Mouse Cells

DTIC Science & Technology

2014-03-01

studies , we show that subtoxic levels of 2-chloroethyl ethylsulfide (2-CEES), a mustard gas analog, also induce centrosome amplification and chromosome...instability in cells, which may hasten the mutation rate necessary for tumorigenesis. These studies offer an explanation why those exposed to mustard...unequally, resulting in chromosome  instability,  a common  phenotype of cancer cells.  In our  studies , we show that subtoxic levels of 2

The RanGTP Pathway: From Nucleo-Cytoplasmic Transport to Spindle Assembly and Beyond

PubMed Central

Cavazza, Tommaso; Vernos, Isabelle

2016-01-01

The small GTPase Ran regulates the interaction of transport receptors with a number of cellular cargo proteins. The high affinity binding of the GTP-bound form of Ran to import receptors promotes cargo release, whereas its binding to export receptors stabilizes their interaction with the cargo. This basic mechanism linked to the asymmetric distribution of the two nucleotide-bound forms of Ran between the nucleus and the cytoplasm generates a switch like mechanism controlling nucleo-cytoplasmic transport. Since 1999, we have known that after nuclear envelope breakdown (NEBD) Ran and the above transport receptors also provide a local control over the activity of factors driving spindle assembly and regulating other aspects of cell division. The identification and functional characterization of RanGTP mitotic targets is providing novel insights into mechanisms essential for cell division. Here we review our current knowledge on the RanGTP system and its regulation and we focus on the recent advances made through the characterization of its mitotic targets. We then briefly review the novel functions of the pathway that were recently described. Altogether, the RanGTP system has moonlighting functions exerting a spatial control over protein interactions that drive specific functions depending on the cellular context. PMID:26793706

Biowaste monitoring system for shuttle

NASA Technical Reports Server (NTRS)

Fogal, G. L.; Sauer, R. L.

1975-01-01

The acquisition of crew biomedical data has been an important task on all manned space missions from Project Mercury through the recently completed Skylab Missions. The monitoring of metabolic wastes from the crew is an important aspect of this activity. On early missions emphasis was placed on the collection and return of biowaste samples for post-mission analysis. On later missions such as Skylab, equipment for inflight measurement was also added. Life Science experiments are being proposed for Shuttle missions which will require the inflight measurement and sampling of metabolic wastes. In order to minimize the crew impact associated with these requirements, a high degree of automation of these processes will be required. This paper reviews the design and capabilities of urine biowaste monitoring equipment provided on past-manned space programs and defines and describes the urine volume measurement and sampling equipment planned for the Shuttle Orbiter program.

First Shuttle/747 Captive Flight

NASA Technical Reports Server (NTRS)

1977-01-01

The Space Shuttle prototype Enterprise rides smoothly atop NASA's first Shuttle Carrier Aircraft (SCA), NASA 905, during the first of the shuttle program's Approach and Landing Tests (ALT) at the Dryden Flight Research Center, Edwards, California, in 1977. During the nearly one year-long series of tests, Enterprise was taken aloft on the SCA to study the aerodynamics of the mated vehicles and, in a series of five free flights, tested the glide and landing characteristics of the orbiter prototype. In this photo, the main engine area on the aft end of Enterprise is covered with a tail cone to reduce aerodynamic drag that affects the horizontal tail of the SCA, on which tip fins have been installed to increase stability when the aircraft carries an orbiter. The Space Shuttle Approach and Landings Tests (ALT) program allowed pilots and engineers to learn how the Space Shuttle and the modified Boeing 747 Shuttle Carrier Aircraft (SCA) handled during low-speed flight and landing. The Enterprise, a prototype of the Space Shuttles, and the SCA were flown to conduct the approach and landing tests at the NASA Dryden Flight Research Center, Edwards, California, from February to October 1977. The first flight of the program consisted of the Space Shuttle Enterprise attached to the Shuttle Carrier Aircraft. These flights were to determine how well the two vehicles flew together. Five 'captive-inactive' flights were flown during this first phase in which there was no crew in the Enterprise. The next series of captive flights was flown with a flight crew of two on board the prototype Space Shuttle. Only three such flights proved necessary. This led to the free-flight test series. The free-flight phase of the ALT program allowed pilots and engineers to learn how the Space Shuttle handled in low-speed flight and landing attitudes. For these landings, the Enterprise was flown by a crew of two after it was released from the top of the SCA. The vehicle was released at altitudes

Space Shuttle Project

NASA Image and Video Library

1992-07-09

As the orbiter Columbia (STS-50) rolled down Runway 33 of Kennedy Space Center's (KSC) Shuttle Landing Facility, its distinctively colored drag chute deployed to slow down the spaceship. This landing marked OV-102's first end-of-mission landing at KSC and the tenth in the program, and the second shuttle landing with the drag chute. Edwards Air Force Base, CA, was the designated prime for the landing of Mission STS-50, but poor weather necessitated the switch to KSC after a one-day extension of the historic flight. STS-50 was the longest in Shuttle program historyo date, lasting 13 days, 19 hours, 30 minutes and 4 seconds. A crew of seven and the USML-1 were aboard.

Human TREX2 components PCID2 and centrin 2, but not ENY2, have distinct functions in protein export and co-localize to the centrosome.

PubMed

Cunningham, Corey N; Schmidt, Casey A; Schramm, Nathaniel J; Gaylord, Michelle R; Resendes, Karen K

2014-01-15

TREX-2 is a five protein complex, conserved from yeast to humans, involved in linking mRNA transcription and export. The centrin 2 subunit of TREX-2 is also a component of the centrosome and is additionally involved in a distinctly different process of nuclear protein export. While centrin 2 is a known multifunctional protein, the roles of other human TREX-2 complex proteins other than mRNA export are not known. In this study, we found that human TREX-2 member PCID2 but not ENY2 is involved in some of the same cellular processes as those of centrin 2 apart from the classical TREX-2 function. PCID2 is present at the centrosome in a subset of HeLa cells and this localization is centrin 2 dependent. Furthermore, the presence of PCID2 at the centrosome is prevalent throughout the cell cycle as determined by co-staining with cyclins E, A and B. PCID2 but not ENY2 is also involved in protein export. Surprisingly, siRNA knockdown of PCID2 delayed the rate of nuclear protein export, a mechanism distinct from the effects of centrin 2, which when knocked down inhibits export. Finally we showed that co-depletion of centrin 2 and PCID2 leads to blocking rather than delaying nuclear protein export, indicating the dominance of the centrin 2 phenotype. Together these results represent the first discovery of specific novel functions for PCID2 other than mRNA export and suggest that components of the TREX-2 complex serve alternative shared roles in the regulation of nuclear transport and cell cycle progression. © 2013 Published by Elsevier Inc.

Shuttle Gaseous Hydrogen Venting Risk from Flow Control Valve Failure

NASA Technical Reports Server (NTRS)

Drummond, J. Philip; Baurle, Robert A.; Gafney, Richard L.; Norris, Andrew T.; Pellett, Gerald L.; Rock, Kenneth E.

2009-01-01

This paper describes a series of studies to assess the potential risk associated with the failure of one of three gaseous hydrogen flow control valves in the orbiter's main propulsion system during the launch of Shuttle Endeavour (STS-126) in November 2008. The studies focused on critical issues associated with the possibility of combustion resulting from release of gaseous hydrogen from the external tank into the atmosphere during assent. The Shuttle Program currently assumes hydrogen venting from the external tank will result in a critical failure. The current effort was conducted to increase understanding of the risk associated with venting hydrogen given the flow control valve failure scenarios being considered in the Integrated In-Flight Anomaly Investigation being conducted by NASA.

Food packages for Space Shuttle

NASA Technical Reports Server (NTRS)

Fohey, M. F.; Sauer, R. L.; Westover, J. B.; Rockafeller, E. F.

1978-01-01

The paper reviews food packaging techniques used in space flight missions and describes the system developed for the Space Shuttle. Attention is directed to bite-size food cubes used in Gemini, Gemini rehydratable food packages, Apollo spoon-bowl rehydratable packages, thermostabilized flex pouch for Apollo, tear-top commercial food cans used in Skylab, polyethylene beverage containers, Skylab rehydratable food package, Space Shuttle food package configuration, duck-bill septum rehydration device, and a drinking/dispensing nozzle for Space Shuttle liquids. Constraints and testing of packaging is considered, a comparison of food package materials is presented, and typical Shuttle foods and beverages are listed.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nishino, Tomonori G.; Department of Biotechnology, University of Tokyo, Bunkyo-ku, Tokyo 113-8657; Miyazaki, Masaya

Class IIa histone deacetylases (HDACs) form complexes with a class of transcriptional repressors in the nucleus. While screening for compounds that could block the association of HDAC4 with the BTB domain-containing transcriptional repressor Bach2, we discovered that phorbol 12-myristate 13-acetate (PMA) induced the cytoplasmic retention of HDAC4 mutants lacking a nuclear export signal (NES). Although PMA treatment and PKD overexpression has been proposed to facilitate the nuclear export of class IIa HDACs by creating 14-3-3 binding sites containing phosphoserines, our experiments using HDAC mutants demonstrated that PMA greatly reduces nuclear import. PMA treatment repressed the NLS activity in a mannermore » dependent on 14-3-3 binding. These results suggest that nuclear HDAC4 is not tethered in the nucleus, but instead shuttles between the nucleus and the cytoplasm. Phosphorylation-induced 14-3-3 binding biases the balance of nucleo-cytoplasmic shuttling toward the cytoplasm by inhibiting nuclear import.« less

Space Shuttle Abort Evolution

NASA Technical Reports Server (NTRS)

Henderson, Edward M.; Nguyen, Tri X.

2011-01-01

This paper documents some of the evolutionary steps in developing a rigorous Space Shuttle launch abort capability. The paper addresses the abort strategy during the design and development and how it evolved during Shuttle flight operations. The Space Shuttle Program made numerous adjustments in both the flight hardware and software as the knowledge of the actual flight environment grew. When failures occurred, corrections and improvements were made to avoid a reoccurrence and to provide added capability for crew survival. Finally some lessons learned are summarized for future human launch vehicle designers to consider.

Shuttle Discovery Fly-Over

NASA Image and Video Library

2012-04-17

Space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) flies over the Steven F. Udvar-Hazy Center, Tuesday, April 17, 2012, in Washington. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Dane Penland)

Space Shuttle Discovery Landing

NASA Image and Video Library

2012-04-17

Space Shuttle Discovery mounted atop a 747 Shuttle Carrier Aircraft (SCA) approaches the runway for landing at Washington Dulles International Airport, Tuesday April 17, 2012, in Sterling, Va. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Paul E. Alers)

Space Shuttle Discovery Landing

NASA Image and Video Library

2012-04-17

Space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) lands at Washington Dulles International Airport, Tuesday, April 17, 2012, in Sterling, Va. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Eric Long)

Shuttle Discovery Mated to 747 SCA

NASA Technical Reports Server (NTRS)

1983-01-01

The Space Shuttle Discovery rides atop '905,' NASA's 747 Shuttle Carrier Aircraft, on its delivery flight from California to the Kennedy Space Center, Florida, where it was prepared for its first orbital mission for 30 August to 5 September 1984. The NASA 747, obtained in 1974, has special support struts atop the fuselage and internal strengthening to accommodate the additional weight of the orbiters. Small vertical fins have also been added to the tips of the horizontal stabilizers for additional stability due to air turbulence on the control surfaces caused by the orbiters. A second modified 747, no. 911, went in to service in November 1990 and is also used to ferry orbiters to destinations where ground transportation is not practical. Space Shuttles are the main element of America's Space Transportation System and are used for space research and other space applications. The shuttles are the first vehicles capable of being launched into space and returning to Earth on a routine basis. Space Shuttles are used as orbiting laboratories in which scientists and mission specialists conduct a wide variety of scientific experiments. Crews aboard shuttles place satellites in orbit, rendezvous with satellites to carry out repair missions and return them to space, and retrieve satellites and return them to Earth for refurbishment and reuse. Space Shuttles are true aerospace vehicles. They leave Earth and its atmosphere under rocket power provided by three liquid-propellant main engines with two solid-propellant boosters attached plus an external liquid-fuel tank. After their orbital missions, they streak back through the atmosphere and land like airplanes. The returning shuttles, however, land like gliders, without power and on runways. Other rockets can place heavy payloads into orbit, but, they can only be used once. Space Shuttles are designed to be continually reused. When Space Shuttles are used to transport complete scientific laboratories into space, the

ENDEAVOR LANDS AT THE SHUTTLE LANDING FACILITY [SLF] ATOP 747 SHUTTLE CARRIER

NASA Technical Reports Server (NTRS)

1991-01-01

The newest addition to the space shuttle orbiter fleet, Endeavour, makes a graceful entrance at KSC atop the 747 Shuttle Carrier aircraft. Next stops for OV-105 are demating from the Boeing Aircraft and extended stays in the VAB and OPF. Endeavour's maiden space flight is scheduled in 1992.

Space Shuttle Payload Information Source

NASA Technical Reports Server (NTRS)

Griswold, Tom

2000-01-01

The Space Shuttle Payload Information Source Compact Disk (CD) is a joint NASA and USA project to introduce Space Shuttle capabilities, payload services and accommodations, and the payload integration process. The CD will be given to new payload customers or to organizations outside of NASA considering using the Space Shuttle as a launch vehicle. The information is high-level in a visually attractive format with a voice over. The format is in a presentation style plus 360 degree views, videos, and animation. Hyperlinks are provided to connect to the Internet for updates and more detailed information on how payloads are integrated into the Space Shuttle.

Spectral characteristics of Shuttle glow

NASA Technical Reports Server (NTRS)

Viereck, R. A.; Mende, S. B.; Murad, E.; Swenson, G. R.; Pike, C. P.; Culbertson, F. L.; Springer, R. C.

1992-01-01

The glowing cloud near the ram surfaces of the Space Shuttle was observed with a hand-held, intensified spectrograph operated by the astronauts from the aft-flight-deck of the Space Shuttle. The spectral measurements were made between 400 and 800 nm with a resolution of 3 nm. Analysis of the spectral response of the instrument and the transmission of the Shuttle window was performed on orbit using earth-airglow OH Meinel bands. This analysis resulted in a correction of the Shuttle glow intensity in the spectral region between 700 and 800 nm. The data presented in this report is in better agreement with laboratory measurements of the NO2 continuum.

TDRSS S-shuttle unique receiver equipment

NASA Astrophysics Data System (ADS)

Weinberg, A.; Schwartz, J. J.; Spearing, R.

1985-01-01

Beginning with STS-9, the Tracking and Date Relay Satellite system (TDRSS) will start providing S- and Ku-band communications and tracking support to the Space Shuttle and its payloads. The most significant element of this support takes place at the TDRSS White Sands Ground Terminal, which processes the Shuttle return link S- and Ku-band signals. While Ku-band hardware available to other TDRSS users is also applied to Ku-Shuttle, stringent S-Shuttle link margins have precluded the application of the standard TDRSS S-band processing equipment to S-Shuttle. It was therfore found necessary to develop a unique S-Shuttle Receiver that embodies state-of-the-art digital technology and processing techniques. This receiver, developed by Motorola, Inc., enhances link margins by 1.5 dB relative to the standard S-band equipment and its bit error rate performance is within a few tenths of a dB of theory. An overview description of the Space Shuttle Receiver Equipment (SSRE) is presented which includes the presentation of block diagrams and salient design features. Selected, measured performance results are also presented.

Space Shuttle Projects

NASA Image and Video Library

1978-09-01

This photograph shows stacking of the left side of the solid rocket booster (SRB) segments in the Dynamic Test Stand at the east test area of the Marshall Space Flight Center (MSFC). Staging shown here are the aft skirt, aft segment, and aft center segment. The SRB was attached to the external tank (ET) and then the orbiter later for the Mated Vertical Ground Vibration Test (MVGVT), that resumed in October 1978. The stacking of a complete Shuttle in the Dynamic Test Stand allowed test engineers to perform ground vibration testing on the Shuttle in its liftoff configuration. The purpose of the MVGVT is to verify that the Space Shuttle would perform as predicted during launch. The platforms inside the Dynamic Test Stand were modified to accommodate two SRB's to which the ET was attached.

Space Shuttle Projects

NASA Image and Video Library

1978-09-01

This photograph shows the left side of the solid rocket booster (SRB) segment as it awaits being mated to the nose cone and forward skirt in the Dynamic Test Stand at the east test area of the Marshall Space Flight Center (MSFC). The SRB would be attached to the external tank (ET) and then the orbiter later for the Mated Vertical Ground Vibration Test (MVGVT), that resumed in October 1978. The stacking of a complete Shuttle in the Dynamic Test Stand allowed test engineers to perform ground vibration testing on the Shuttle in its liftoff configuration. The purpose of the MVGVT was to verify that the Space Shuttle would perform as predicted during launch. The platforms inside the Dynamic Test Stand were modified to accommodate two SRB's to which the ET was attached.

Space Shuttle Projects

NASA Image and Video Library

1978-09-01

Workmen in the Dynamic Test Stand lowered the nose cone into place to complete stacking of the left side of the solid rocket booster (SRB) in the Dynamic Test Stand at the east test area of the Marshall Space Flight Center (MSFC). The SRB would be attached to the external tank (ET) and then the orbiter later for the Mated Vertical Ground Vibration Test (MVGVT), that resumed in October 1978. The stacking of a complete Shuttle in the Dynamic Test Stand allowed test engineers to perform ground vibration testing on the Shuttle in its liftoff configuration. The purpose of the MVGVT was to verify that the Space Shuttle would perform as predicted during launch. The platforms inside the Dynamic Test Stand were modified to accommodate two SRB'S to which the ET was attached.

Managing Toxicological Risks: The Legacy of Shuttle Operations

NASA Technical Reports Server (NTRS)

James, John T.

2011-01-01

Space toxicology greatly matured as a result of research and operations associated with the Shuttle. Materials offgassing had been a manageable concern since the Apollo days, but we learned to pay careful attention to compounds that could escape containment, to combustion events, to toxic propellants, to overuse of utility compounds, and to microbial and human metabolites. We also learned that flying real-time hardware to monitor air pollutants was a pathway with unanticipated speed bumps. Each new orbiter was tested for any excess offgassing products that could pollute the air during flight. In the late 1990s toxicologists and safety experts developed a 5-level toxicity rating system to guide containment of toxic compounds. This system is now in use aboard the International Space Station (ISS). Several combustion events during Shuttle Mir and also during Shuttle free-flight impelled toxicologists to identify hardware capable of monitoring toxic products; however, rapid adaptation of the hardware for the unique conditions of spaceflight caused unexpected missteps. Current and planned combustion analyzers would be useful to commercial partners that wish to manage the risk of health effects from thermal events. Propellants received special attention during the Shuttle program because of the possibility of bringing them into the habitable volume on extravehicular activity suits. Monitors for the airlocks were developed to mitigate this risk. Utility materials, such as lubricants, posed limited toxicological problems because water was not recovered. One clearly documented case of microbial metabolites polluting the Shuttle atmosphere was noted, and this has implications for commercial flights and control of microbes. Finally, carbon dioxide, the major human metabolite, episodically presented air quality problems aboard Shuttle, especially when nominal air flows were obstructed. Commercial vehicles must maintain robust air circulation given the anticipated high density

Space Shuttle redesign status

NASA Technical Reports Server (NTRS)

Brand, Vance D.

1986-01-01

NASA has conducted an extensive redesign effort for the Space Shutle in the aftermath of the STS 51-L Challenger accident, encompassing not only Shuttle vehicle and booster design but also such system-wide factors as organizational structure, management procedures, flight safety, flight operations, sustainable flight rate, and maintenance safeguards. Attention is presently given to Solid Rocket Booster redesign features, the Shuttle Main Engine's redesigned high pressure fuel and oxidizer turbopumps, the Shuttle Orbiter's braking and rollout (landing gear) system, the entry control mode of the flight control system, a 'split-S' abort maneuver for the Orbiter, and crew escape capsule proposals.

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

A set of cranes and wind restraints constructed to remove space shuttle Enterprise from atop NASA's 747 Shuttle Carrier Aircraft are being put into place at John F. Kennedy (JFK) International Airport in New York, Saturday, May 12, 2012. Enterprise will be placed on a barge that will move by tugboat up the Hudson River to Intrepid in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Discovery Landing

NASA Image and Video Library

2012-04-17

Space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) taxis in front of the main terminal at Washington Dulles International Airport, Tuesday, April 17, 2012, in Sterling, Va. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Eric Long)

MSFC shuttle lightning research

NASA Technical Reports Server (NTRS)

Vaughan, Otha H., Jr.

1993-01-01

The shuttle mesoscale lightning experiment (MLE), flown on earlier shuttle flights, and most recently flown on the following space transportation systems (STS's), STS-31, -32, -35, -37, -38, -40, -41, and -48, has continued to focus on obtaining additional quantitative measurements of lightning characteristics and to create a data base for use in demonstrating observation simulations for future spaceborne lightning mapping systems. These flights are also providing design criteria data for the design of a proposed shuttle MLE-type lightning research instrument called mesoscale lightning observational sensors (MELOS), which are currently under development here at MSFC.

Problem-Solving Test: Nucleocytoplasmic Shuttling of Pre-mRNA Binding Proteins

ERIC Educational Resources Information Center

Szeberenyi, Jozsef

2012-01-01

Terms to be familiar with before you start to solve the test: transcription, pre-mRNA, RNA processing, RNA transport, RNA polymerase II, direct and indirect immunofluorescence staining, cell fractionation by centrifugation, oligo(dT)-cellulose chromatography, washing and elution of the column, ribonuclease, SDS-polyacrylamide gel electrophoresis,…

Nucleocytoplasmic trafficking of Nipah virus W protein involves multiple discrete interactions with the nuclear import and export machinery.

PubMed

Audsley, Michelle D; Jans, David A; Moseley, Gregory W

2016-10-21

Paramyxoviruses replicate in the cytoplasm with no obvious requirement to interact with the nucleus. Nevertheless, the W protein of the highly lethal bat-borne paramyxovirus Nipah virus (NiV) is known to undergo specific targeting to the nucleus, mediated by a single nuclear localisation signal (NLS) within the C-terminal domain. Here, we report for the first time that additional sites modulate nucleocytoplasmic localisation of W. We show that the N-terminal domain interacts with importin α1 and contributes to nuclear accumulation of W, indicative of a novel N-terminal NLS. We also find that W undergoes exportin-1 mediated nuclear export, dependent on a leucine at position 174. Together, these data enable significant revision of the generally accepted model of W trafficking, with implications for understanding of the mechanisms of NiV immune evasion. Copyright © 2016 Elsevier Inc. All rights reserved.

Space shuttle launch vehicle aerodynamic uncertainties: Lessons learned

NASA Technical Reports Server (NTRS)

Hamilton, J. T.

1983-01-01

The chronological development and evolution of an uncertainties model which defines the complex interdependency and interaction of the individual Space Shuttle element and component uncertainties for the launch vehicle are presented. Emphasis is placed on user requirements which dictated certain concessions, simplifications, and assumptions in the analytical model. The use of the uncertainty model in the vehicle design process and flight planning support is discussed. The terminology and justification associated with tolerances as opposed to variations are also presented. Comparisons of and conclusions drawn from flight minus predicted data and uncertainties are given. Lessons learned from the Space Shuttle program concerning aerodynamic uncertainties are examined.

Space Shuttle Projects

NASA Image and Video Library

1976-01-01

This is a cutaway illustration of the Space Shuttle external tank (ET) with callouts. The giant cylinder, higher than a 15-story building, with a length of 154-feet (47-meters) and a diameter of 27.5-feet (8.4-meters), is the largest single piece of the Space Shuttle. During launch, the ET also acts as a backbone for the orbiter and solid rocket boosters. Separate pressurized tank sections within the external tank hold the liquid hydrogen fuel and liquid oxygen oxidizer for the Shuttle's three main engines. During launch, the ET feeds the fuel under pressure through 17-inch (43.2-centimeter) ducts that branch off into smaller lines that feed directly into the main engines. The main engines consume 64,000 gallons (242,260 liters) of fuel each minute. Machined from aluminum alloys, the Space Shuttle's external tank is currently the only part of the launch vehicle that is not reused. After its 526,000-gallons (1,991,071 liters) of propellants are consumed during the first 8.5-minutes of flight, it is jettisoned from the orbiter and breaks up in the upper atmosphere, its pieces falling into remote ocean waters. The Marshall Space Flight Center was responsible for developing the ET.

Space Shuttle Projects

NASA Image and Video Library

1995-11-12

The STS-76 crew patch depicts the Space Shuttle Atlantis and Russia's Mir Space Station as the space ships prepare for a rendezvous and docking. The Spirit of 76, an era of new beginnings, is represented by the Space Shuttle rising through the circle of 13 stars in the Betsy Ross flag. STS-76 begins a new period of international cooperation in space exploration with the first Shuttle transport of a United States astronaut, Shannon W. Lucid, to the Mir Space Station for extended joint space research. Frontiers for future exploration are represented by stars and the planets. The three gold trails and the ring of stars in union form the astronaut logo. Two suited extravehicular activity (EVA) crew members in the outer ring represent the first EVA during Shuttle-Mir docked operations. The EVA objectives were to install science experiments on the Mir exterior and to develop procedures for future EVA's on the International Space Station. The surnames of the crew members encircle the patch: Kevin P. Chilton, mission commander; Richard A. Searfoss, pilot; Ronald M. Sega, Michael R. ( Rich) Clifford, Linda M. Godwin and Lucid, all mission specialists. This patch was designed by Brandon Clifford, age 12, and the crew members of STS-76.

Shuttle: forever young?

PubMed

Sietzen, Frank

2002-01-01

NASA has started a 4-phase program of upgrades designed to increase safety and extend use of the space shuttles through the year 2020. Phase I is aimed at improving vehicle safety and supporting the space station. Phase II is aimed at combating obsolescence and includes a checkout launch and control system and protection from micrometeoroids and orbital debris. Phase III is designed to expand or enhance the capabilities of the shuttle and includes development of an auxiliary power unit, avionics, a channel-wall nozzle, extended nose landing gear, long-life fuel cells, a nontoxic orbital maneuvering system/reaction control system, and a water membrane evaporator. Phase IV is aimed at design of system changes that would alter the shuttle mold line and configuration; projects include a five-segment solid rocket booster, liquid flyback boosters, and a crew escape module.

Space Shuttle Projects

NASA Image and Video Library

1992-09-01

The STS-53 crew portrait included astronauts (front left to right): Guion S. Bluford, and James S. Voss, mission specialists. On the back row, left to right, are David M. Walker, commander; Robert D. Cabana, Pilot; and Michael R. (Rick) Clifford, mission specialist. The crew launched aboard the Space Shuttle Discovery on December 2, 1992 at 8:24:00 am (EST). This mission marked the final classified shuttle flight for the Department of Defense (DOD).

Space Shuttle Project

NASA Image and Video Library

1995-03-18

The Space Shuttle Endeavour (STS-67) lands at Edwards Air Force Base in southern California after successfully completing NASA's longest plarned shuttle mission. The seven-member crew conducted round-the-clock observations with the ASTRO-2 observatory, a trio of telescopes designed to study the universe of ultraviolet astronomy. Because of Earth's protective ozone layer ultraviolet light from celestial objects does not reach gound-based telescopes, and such studies can only be conducted from space.

Representative shuttle evaporative heat sink

NASA Technical Reports Server (NTRS)

Hixon, C. W.

1978-01-01

The design, fabrication, and testing of a representative shuttle evaporative heat sink (RSEHS) system which vaporizes an expendable fluid to provide cooling for the shuttle heat transport fluid loop is reported. The optimized RSEHS minimum weight design meets or exceeds the shuttle flash evaporator system requirements. A cold trap which cryo-pumps flash evaporator exhaust water from the CSD vacuum chamber test facility to prevent water contamination of the chamber pumping equipment is also described.

Chemical Shuttle Additives in Lithium Ion Batteries

DOE Office of Scientific and Technical Information (OSTI.GOV)

Patterson, Mary

2013-03-31

The goals of this program were to discover and implement a redox shuttle that is compatible with large format lithium ion cells utilizing LiNi{sub 1/3}Mn{sub 1/3}Co{sub 1/3}O{sub 2} (NMC) cathode material and to understand the mechanism of redox shuttle action. Many redox shuttles, both commercially available and experimental, were tested and much fundamental information regarding the mechanism of redox shuttle action was discovered. In particular, studies surrounding the mechanism of the reduction of the oxidized redox shuttle at the carbon anode surface were particularly revealing. The initial redox shuttle candidate, namely 2-(pentafluorophenyl)-tetrafluoro-1,3,2-benzodioxaborole (BDB) supplied by Argonne National Laboratory (ANL, Lemont,more » Illinois), did not effectively protect cells containing NMC cathodes from overcharge. The ANL-RS2 redox shuttle molecule, namely 1,4-bis(2-methoxyethoxy)-2,5-di-tert-butyl-benzene, which is a derivative of the commercially successful redox shuttle 2,5-di-tert-butyl-1,4-dimethoxybenzene (DDB, 3M, St. Paul, Minnesota), is an effective redox shuttle for cells employing LiFePO{sub 4} (LFP) cathode material. The main advantage of ANL-RS2 over DDB is its larger solubility in electrolyte; however, ANL-RS2 is not as stable as DDB. This shuttle also may be effectively used to rebalance cells in strings that utilize LFP cathodes. The shuttle is compatible with both LTO and graphite anode materials although the cell with graphite degrades faster than the cell with LTO, possibly because of a reaction with the SEI layer. The degradation products of redox shuttle ANL-RS2 were positively identified. Commercially available redox shuttles Li{sub 2}B{sub 12}F{sub 12} (Air Products, Allentown, Pennsylvania and Showa Denko, Japan) and DDB were evaluated and were found to be stable and effective redox shuttles at low C-rates. The Li{sub 2}B{sub 12}F{sub 12} is suitable for lithium ion cells utilizing a high voltage cathode (potential that

STS-92 - Towing of Shuttle Discovery and Boeing 747 Shuttle Carrier Aircraft (SCA)

NASA Technical Reports Server (NTRS)

2000-01-01

The Space Shuttle Discovery sits atop one of NASA's modified Boeing 747 Shuttle Carrier Aircraft as the unusual piggyback duo is towed along a taxiway at NASA's Dryden Flight Research Center at Edwards, California. The Discovery was ferried from NASA Dryden to NASA's Kennedy Space Center in Florida on November 2, 2000, after extensive pre-ferry servicing and preparations. STS-92 was the 100th mission since the fleet of four Space Shuttles began flying in 1981. (Due to schedule changes, missions are not always launched in the order that was originally planned.) The almost 13-day mission, the 46th Shuttle mission to land at Edwards, was the last construction mission for the International Space Station prior to the first scientists taking up residency in the orbiting space laboratory the following month. The seven-member crew on STS-92 included mission specialists Koichi Wakata, Michael Lopez-Alegria, Jeff Wisoff, Bill McArthur and Leroy Chiao, pilot Pam Melroy and mission commander Brian Duffy.

Space Shuttle Enterprise Demate

NASA Image and Video Library

2012-05-12

The space shuttle Enterprise, mounted on transport vehicle, is backed into a temporary hanger after being demated from the NASA 747 Shuttle Carrier Aircraft (SCA) at John F. Kennedy (JFK) International Airport in Jamica, New York, Sunday, May 13, 2012. Enterprise will be placed on a barge that will move by tugboat up the Hudson River to the Intrepid Sea, Air & Space Museum in June. The shuttle will be lifted by crane and placed on the flight deck of the Intrepid, where it will be on exhibit to the public starting this summer in a temporary climate-controlled pavilion. Photo Credit: (NASA/Kim Shiflet)

Space Shuttle Discovery Landing

NASA Image and Video Library

2012-04-17

Space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) lands at Washington Dulles International Airport, Tuesday, April 17, 2012, in Sterling, Va. The Steven F. Udvar-Hazy Center is seen in the background. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Eric Long)

Separate responses of karyopherins to glucose and amino acid availability regulate nucleocytoplasmic transport

PubMed Central

Huang, Hsiao-Yun; Hopper, Anita K.

2014-01-01

The importin-β family members (karyopherins) mediate the majority of nucleocytoplasmic transport. Msn5 and Los1, members of the importin-β family, function in tRNA nuclear export. tRNAs move bidirectionally between the nucleus and the cytoplasm. Nuclear tRNA accumulation occurs upon amino acid (aa) or glucose deprivation. To understand the mechanisms regulating tRNA subcellular trafficking, we investigated whether Msn5 and Los1 are regulated in response to nutrient availability. We provide evidence that tRNA subcellular trafficking is regulated by distinct aa-sensitive and glucose-sensitive mechanisms. Subcellular distributions of Msn5 and Los1 are altered upon glucose deprivation but not aa deprivation. Redistribution of tRNA exportins from the nucleus to the cytoplasm likely provides one mechanism for tRNA nuclear distribution upon glucose deprivation. We extended our studies to other members of the importin-β family and found that all tested karyopherins invert their subcellular distributions upon glucose deprivation but not aa deprivation. Glucose availability regulates the subcellular distributions of karyopherins likely due to alteration of the RanGTP gradient since glucose deprivation causes redistribution of Ran. Thus nuclear–cytoplasmic distribution of macromolecules is likely generally altered upon glucose deprivation due to collapse of the RanGTP gradient and redistribution of karyopherins between the nucleus and the cytoplasm. PMID:25057022

Nuclear Shuttle Logistics Configuration

NASA Technical Reports Server (NTRS)

1971-01-01

This 1971 artist's concept shows the Nuclear Shuttle in both its lunar logistics configuraton and geosynchronous station configuration. As envisioned by Marshall Space Flight Center Program Development persornel, the Nuclear Shuttle would deliver payloads to lunar orbits or other destinations then return to Earth orbit for refueling and additional missions.

Immunosuppressive Yersinia Effector YopM Binds DEAD Box Helicase DDX3 to Control Ribosomal S6 Kinase in the Nucleus of Host Cells.

PubMed

Berneking, Laura; Schnapp, Marie; Rumm, Andreas; Trasak, Claudia; Ruckdeschel, Klaus; Alawi, Malik; Grundhoff, Adam; Kikhney, Alexey G; Koch-Nolte, Friedrich; Buck, Friedrich; Perbandt, Markus; Betzel, Christian; Svergun, Dmitri I; Hentschke, Moritz; Aepfelbacher, Martin

2016-06-01

Yersinia outer protein M (YopM) is a crucial immunosuppressive effector of the plaque agent Yersinia pestis and other pathogenic Yersinia species. YopM enters the nucleus of host cells but neither the mechanisms governing its nucleocytoplasmic shuttling nor its intranuclear activities are known. Here we identify the DEAD-box helicase 3 (DDX3) as a novel interaction partner of Y. enterocolitica YopM and present the three-dimensional structure of a YopM:DDX3 complex. Knockdown of DDX3 or inhibition of the exportin chromosomal maintenance 1 (CRM1) increased the nuclear level of YopM suggesting that YopM exploits DDX3 to exit the nucleus via the CRM1 export pathway. Increased nuclear YopM levels caused enhanced phosphorylation of Ribosomal S6 Kinase 1 (RSK1) in the nucleus. In Y. enterocolitica infected primary human macrophages YopM increased the level of Interleukin-10 (IL-10) mRNA and this effect required interaction of YopM with RSK and was enhanced by blocking YopM's nuclear export. We propose that the DDX3/CRM1 mediated nucleocytoplasmic shuttling of YopM determines the extent of phosphorylation of RSK in the nucleus to control transcription of immunosuppressive cytokines.

Space Shuttle operational logistics plan

NASA Technical Reports Server (NTRS)

Botts, J. W.

1983-01-01

The Kennedy Space Center plan for logistics to support Space Shuttle Operations and to establish the related policies, requirements, and responsibilities are described. The Directorate of Shuttle Management and Operations logistics responsibilities required by the Kennedy Organizational Manual, and the self-sufficiency contracting concept are implemented. The Space Shuttle Program Level 1 and Level 2 logistics policies and requirements applicable to KSC that are presented in HQ NASA and Johnson Space Center directives are also implemented.

Space Shuttle Project

NASA Image and Video Library

1993-04-17

A four-million-mile journey draws to a flawless ending as the orbiter Discovery (STS-56) lands at Kennedy Space Center's (KSC) Shuttle Landing Facility. Aboard for the second shuttle mission of 1993 were a crew of five and the Atmospheric Laboratory for Applications and Science 2 (ATLAS 2), the second in a series of missions to study the sun's energy output and Earth's middle atmosphere chemical make-up, and how these factors affect levels of ozone.

Space Shuttle Project

NASA Image and Video Library

1992-09-12

A smooth countdown culminated in a picture-perfect launch as the Space Shuttle Endeavour (STS-47) climbed skyward atop a ladder of billowing smoke. Primary payload for the plarned seven-day flight was Spacelab-J science laboratory. The second flight of Endeavour marks a number of historic firsts: the first space flight of an African-American woman, the first Japanese citizen to fly on a Space Shuttle, and the first married couple to fly in space.

Space Shuttle Discovery Launch

NASA Image and Video Library

2008-05-31

NASA Administrator, Michael Griffin watches the launch of the Space Shuttle Discovery (STS-124) from the Launch Control Center Saturday, May 31, 2008, at the Kennedy Space Center in Cape Canaveral, Fla. The Shuttle lifted off from launch pad 39A at 5:02 p.m. EDT. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Projects

NASA Image and Video Library

1991-08-01

The free-flying Tracking and Data Relay Satellite-E (TDRS-E), still attached to an Inertial Upper Stage (IUS), was photographed by one of the crewmembers during the STS-43 mission. The TDRS-E was boosted by the IUS into geosynchronous orbit and positioned to remain stationary 22,400 miles above the Pacific Ocean southwest of Hawaii. The TDRS system provides almost uninterrupted communications with Earth-orbiting Shuttles and satellites, and had replaced the intermittent coverage provided by globe-encircling ground tracking stations used during the early space program. The TDRS can transmit and receive data, and track a user spacecraft in a low Earth orbit. The IUS is an unmarned transportation system designed to ferry payloads from low Earth orbit to higher orbits that are unattainable by the Shuttle. The Space Shuttle Orbiter Atlantis for the STS-43 mission was launched on August 2, 1991.

Space Shuttle Projects

NASA Image and Video Library

1991-08-01

The primary payload of the STS-43 mission, Tracking and Data Relay Satellite-E (TDRS-E) attached to an Inertial Upper Stage (IUS) was photographed at the moment of its release from the cargo bay of the Space Shuttle Orbiter Atlantis. The TDRS-E was boosted by the IUS into geosynchronous orbit and positioned to remain stationary 22,400 miles above the Pacific Ocean southwest of Hawaii. The TDRS system provides almost uninterrupted communications with Earth-orbiting Shuttles and satellites, and had replaced the intermittent coverage provided by globe-encircling ground tracking stations used during the early space program. The TDRS can transmit and receive data, and track a user spacecraft in a low Earth orbit. The IUS is an unmarned transportation system designed to ferry payloads from low Earth orbit to higher orbits that are unattainable by the Shuttle. The launch of STS-43 occurred on August 2, 1991.

Space shuttle propulsion systems on-board checkout and monitoring system development study (extension). Volume 2: Guidelines for for incorporation of the onboard checkout and monitoring function on the space shuttle

NASA Technical Reports Server (NTRS)

1972-01-01

Guidelines are presented for incorporation of the onboard checkout and monitoring function (OCMF) into the designs of the space shuttle propulsion systems. The guidelines consist of and identify supporting documentation; requirements for formulation, implementation, and integration of OCMF; associated compliance verification techniques and requirements; and OCMF terminology and nomenclature. The guidelines are directly applicable to the incorporation of OCMF into the design of space shuttle propulsion systems and the equipment with which the propulsion systems interface. The techniques and general approach, however, are also generally applicable to OCMF incorporation into the design of other space shuttle systems.

XCTK1: A Xenopus C-terminal Kinesin-like Protein

NASA Technical Reports Server (NTRS)

Winfree, Seth; Wilhelm, Heike; Sawyer, Alan; Karsenti, Eric; Mitchison, Tim; Walczak, Claire; Reinsch, Sigrid; Dalton, Bonnie (Technical Monitor)

2002-01-01

XCTK1 is 97kDa kinesin-like protein homologous to FKIF2 and KIFC3. XCTK1 is present at picomolar levels in eggs, embryos and cultured cells in a soluble high-molecular weight complex that is not associated with membranes. XCKT1 localizes to centrosomes in Xenopus A6 cells. Anti-XCTK1 antibodies also localize to spindle poles when injected into A6 cells or when added to extracts during in vitro spindle assembly reactions. XCTK1 is associated with the center of taxol-induced microtubule asters in extracts. Therefore its localization to poles is dependent on microtubule minus-ends and not on centrosomes per se. Overexpression of XCTK1 leads to centrosome destruction in cultured cells. XCTK1 was tagged at either the N- or C-terminus and transfected into Xenopus A6 cells At low expression levels, XCTK1 associated with centrosomes. At higher levels, the protein localized to insoluble cytoplasmic structures. Gamma-tubulin staining was dramatically decreased from centrosomes or altogether absent. The centrosomal SPJ antigen colocalized with XCTK1-containing structures. Upon nocodozole treatment, microtubules failed to regrow from the centrosomes indicating that overexpression of XCTK1 severely compromises centrosomal function. Current studies are aimed at determining whether XCTK1 interacts directly with centrosomal proteins and to determine the effects of XCTK1 depletion on oocyte maturation and embryogenesis.

Space Shuttle Projects

NASA Image and Video Library

1983-07-01

This photograph was taken during the final assembly phase of the Space Shuttle light weight external tanks (LWT) 5, 6, and 7 at the Michoud Assembly Facility in New Orleans, Louisiana. The giant cylinder, higher than a 15-story building, with a length of 154-feet (47-meters) and a diameter of 27.5-feet (8.4-meters), is the largest single piece of the Space Shuttle. During launch, the external tank (ET) acts as a backbone for the orbiter and solid rocket boosters. In separate, internal pressurized tank sections, the ET holds the liquid hydrogen fuel and liquid oxygen oxidizer for the Shuttle's three main engines. During launch, the ET feeds the fuel under pressure through 17-inch (43.2-centimeter) ducts which branch off into smaller lines that feed directly into the main engines. Some 64,000 gallons (242,260 liters) of fuel are consumed by the main engines each minute. Machined from aluminum alloys, the Space Shuttle's ET is the only part of the launch vehicle that currently is not reused. After its 526,000 gallons (1,991,071 liters) of propellants are consumed during the first 8.5 minutes of flight, it is jettisoned from the orbiter and breaks up in the upper atmosphere, its pieces falling into remote ocean waters. The Marshall Space Flight Center was responsible for developing the ET

Space Shuttle Projects

NASA Image and Video Library

1989-03-01

This STS-29 mission onboard photo depicts the External Tank (ET) falling toward the ocean after separation from the Shuttle orbiter Discovery. The giant cylinder, higher than a 15-story building, with a length of 154-feet (47-meters) and a diameter of 27,5-feet (8.4-meters), is the largest single piece of the Space Shuttle. During launch, the ET also acts as a backbone for the orbiter and solid rocket boosters. In separate, internal pressurized tank sections, the ET holds the liquid hydrogen fuel and liquid oxygen oxidizer for the Shuttle's three main engines. During launch, the ET feeds the fuel under pressure through 17-inch (43.2-centimeter) ducts which branch off into smaller lines that feed directly into the main engines. Some 64,000 gallons (242,260 liters) of fuel are consumed by the main engines each minute. Machined from aluminum alloys, the Space Shuttle's ET is the only part of the launch vehicle that currently is not reused. After its 526,000 gallons (1,991,071 liters) of propellants are consumed during the first 8.5 minutes of flight, it is jettisoned from the orbiter and breaks up in the upper atmosphere, its pieces falling into remote ocean waters. The Marshall Space Flight Center was responsible for developing the ET.

Space Shuttle Project

NASA Image and Video Library

1992-08-24

A crewmember aboard the Space Shuttle Orbiter Atlantis (STS-46) used a 70mm handheld camera to capture this medium closeup view of early operations with the Tethered Satellite System (TSS). TSS-1 is being deployed from its boom as it is perched above the cargo bay of the Earth-orbiting Shuttle circling the Earth at an altitude of 296 kilometers (184 miles), the TSS-1 will be well within the tenuous, electrically charged layer of the atmosphere known as the ionosphere. There, a satellite attached to the orbiter by a thin conducting cord, or tether, will be reeled from the Shuttle payload bay. On this mission the satellite was plarned to be deployed 20 kilometers (12.5 miles) above the Shuttle. The conducting tether will generate high voltage and electrical currents as it moves through the atmosphere allowing scientists to examine the electrodynamics of a conducting tether system. These studies will not only increase our understanding of physical processes in the near-Earth space environment, but will also help provide an explanation for events witnessed elsewhere in the solar system. The crew of the STS-46 mission were unable to reel the satellite as planned. After several unsuccessful attempts, they were only able to extend the satellite 9.8 kilometers (6.1 miles). The TSS was a cooperative development effort by the Italian Space Agency (ASI), and NASA.

STS-92 - Shuttle Carrier Aircraft (SCA)

NASA Image and Video Library

2000-10-29

One of NASA’s two modified Boeing 747 Shuttle Carrier Aircraft is bathed in the morning Sun at NASA’s Dryden Flight Research Center at Edwards, California. The modified jumbo jetliners are used to ferry the Space Shuttle orbiters between Dryden and the Kennedy Space Center in Florida and Boeing’s Reusable Space Systems modification facility at Palmdale, California. Features which distinguish the two SCAs from standard 747 jetliners are three struts, with associated interior structural strengthening, which protrude from the top of the fuselage (two aft, one forward) on which the orbiter is attached, and two additional vertical stabilizers, one on each end of the standard horizontal stabilizer, to enhance directional stability. All interior furnishings and equipment aft of the forward No. 1 doors have also been removed to reduce weight. The two SCAs are under the operational control of NASA's Johnson Space Center, Houston, Texas.

Space Shuttle Placement Announcement

NASA Image and Video Library

2011-04-12

From left, Pilot of the first space shuttle mission, STS-1, Bob Crippen, NASA Administrator Charles Bolden, NASA Johnson Space Center Director of Flight Crew Operations, and Astronaut, Janet Kavandi, NASA Kennedy Space Center Director and former astronaut Bob Cabana, and Endeavour Vehicle Manager for United Space Alliance Mike Parrish pose for a photograph outside of the an Orbiter Processing Facility with the space shuttle Atlantis shortly after Bolden announced where four space shuttle orbiters will be permanently displayed at the conclusion of the Space Shuttle Program, Tuesday, April 12, 2011, at Kennedy Space Center in Cape Canaveral, Fla. The four orbiters, Enterprise, which currently is on display at the Smithsonian's Steven F. Udvar-Hazy Center near Washington Dulles International Airport, will move to the Intrepid Sea, Air & Space Museum in New York, Discovery will move to Udvar-Hazy, Endeavour will be displayed at the California Science Center in Los Angeles and Atlantis, in background, will be displayed at the Kennedy Space Center Visitor’s Complex. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Discovery Landing

NASA Image and Video Library

2012-04-17

NASA Deputy Administrator Lori Garver, at podium, speaks to those in attendance at Apron W after the 747 Shuttle Carrier Aircraft (SCA) with space shuttle Discovery mounted on top rolled to a halt at Washington Dulles International Airport, Tuesday, April 17, 2012 in Sterling, Va. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Dane Penland)

Shuttle Imaging Radar - Geologic applications

NASA Technical Reports Server (NTRS)

Macdonald, H.; Bridges, L.; Waite, W.; Kaupp, V.

1982-01-01

The Space Shuttle, on its second flight (November 12, 1981), carried the first science and applications payload which provided an early demonstration of Shuttle's research capabilities. One of the experiments, the Shuttle Imaging Radar-A (SIR-A), had as a prime objective to evaluate the capability of spaceborne imaging radars as a tool for geologic exploration. The results of the experiment will help determine the value of using the combination of space radar and Landsat imagery for improved geologic analysis and mapping. Preliminary analysis of the Shuttle radar imagery with Seasat and Landsat imagery from similar areas provides evidence that spaceborne radars can significantly complement Landsat interpretation, and vastly improve geologic reconnaissance mapping in those areas of the world that are relatively unmapped because of perpetual cloud cover.

Restartable solid motor stage for shuttle applications

NASA Technical Reports Server (NTRS)

Rohrbaugh, D. J.

1973-01-01

The application of restartable solid motor stages to shuttle missions has been shown to provide a viable supplement to the shuttle program. Restartable solid motors in the 3000 pound class provide a small expendable transfer stage that reduces the demand on the shuttle for the lower energy missions. Shuttle operational requirements and preliminary performance data provided an input for defining design features required for restartable solid motor applications. These data provided a basis for a configuration definition that is compatible with shuttle operations. Mission by mission analysis showed the impact on a NASA supplied mission model. The results showed a 15% reduction in the number of shuttle flights required. In addition the amount of shuttle capability used to complete the mission objectives was significantly reduced. For example, in the 1979 missions there was a 62% reduction in shuttle capability used. The study also showed that the solid motor could provide a supplement to the TUG that would allow TUGS to be used in a recoverable rather than an expendable mode. The study shows a 71% reduction in the number of TUGs that would be expended.

Space Shuttle Projects

NASA Image and Video Library

1984-10-01

The Space Shuttle Discovery en route to Earth orbit for NASA's 51-A mission is reminiscent of a soaring Eagle. The red and white trailing stripes and the blue background, along with the presence of the Eagle, generate memories of America's 208 year-old history and traditions. The two satellites orbiting the Earth backgrounded amidst a celestial scene are a universal representation of the versatility of the Space Shuttle. White lettering against the blue border lists the surnames of the five-member crew.

Space Shuttle Projects

NASA Image and Video Library

1978-09-29

This photo depicts the installation of an External Tank (ET) into the Marshall Space Flight Center Dynamic Test Stand, building 4550. It is being mated to the Solid Rocket Boosters (SRB's) for a Mated Vertical Ground Vibration Test (MVGVT). At 154-feet long and more than 27-feet in diameter, the ET is the largest component of the Space Shuttle, the structural backbone of the entire Shuttle system, and is the only part of the vehicle that is not reusable.

Space Shuttle Project

NASA Image and Video Library

1992-09-12

A smooth countdown culminated in a picture-perfect launch as the Space Shuttle Orbiter Endeavour (STS-47) climbed skyward atop a ladder of billowing smoke on September 12, 1992. The primary payload for the plarned seven-day flight was the Spacelab-J science laboratory. The second flight of Endeavour marks a number of historic firsts: the first space flight of an African-American woman, the first Japanese citizen to fly on a Space Shuttle, and the first married couple to fly in space.

Space Shuttle Projects

NASA Image and Video Library

1985-05-30

The crewmembers of Space Shuttle mission 51-F have chosen as their insignia this design by Houston artist Skip Bradley. The Space Shuttle Challenger is depicted ascending toward the heavens in search of new knowledge in the field of solar and steallar astronomy, with its Spacelab 2 payload. The constellations Leo and Orion are in the positions they will be in, relative to the sun during the flight. The nineteen stars signify that this will be the 19th STS flight.

Space Shuttle Environmental Effects: The First 5 Flights

NASA Technical Reports Server (NTRS)

Potter, A. (Editor)

1983-01-01

Environmental effects associated with the first five Space Shuttle flights were monitored by the National Aeronautics and Space Administration (NASA) and the U.S. Air Force (USAF). Results and interpretations from this effort were reported at the December 1982 joint NASA-USAF conference. The conference proceedings are presented in this document. Most of the monitoring activity was focused on the launch cloud, emphasizing surface effects on the biota and air quality, model prediction of surface concentrations of HCl gas and Al2O3 dust, and airborne measurements of cloud composition. In general, assessments and predictions made in the April 1978 Final Environmental Impact Statement for the Space Shuttle Program were verified. Fallout of acidic mist and dust within 3 mi to 5 mi of the launch pad was the only unexpected effect of the launch. Atomization of deluge water in the Shuttle exhaust is considered to be the most probable cause of this effect. Sonic booms were monitored for several landings at Edwards Air Force Base, California; results agreed well with model predictions.

Environmental protection requirements for scout/shuttle auxiliary stages

NASA Technical Reports Server (NTRS)

Qualls, G. L.; Kress, S. S.; Storey, W. W.; Ransdell, P. N.

1980-01-01

The requirements for enabling the Scout upper stages to endure the expected temperature, mechanical shock, acoustical and mechanical vibration environments during a specified shuttle mission were determined. The study consisted of: determining a shuttle mission trajectory for a 545 kilogram (1200 pound) Scout payload; compilation of shuttle environmental conditions; determining of Scout upper stages environments in shuttle missions; compilation of Scout upper stages environmental qualification criteria and comparison to shuttle mission expected environments; and recommendations for enabling Scout upper stages to endure the exptected shuttle mission environments.

Modification of Experimental Protocols for a Space Shuttle Flight and Applications for the Analysis of Cytoskeletal Structures During Fertilization, Cell Division , and Development in Sea Urchin Embryos

NASA Technical Reports Server (NTRS)

Chakrabarti, Amitabha; Stoecker, Andrew; Schatten, Heide

1995-01-01

To explore the role of microgravity on cytoskeletal organization and skeletal calcium deposition during fertilization, cell division, and early development, the sea urchin was chosen as a model developmental system. Methods were developed to employ light, immunofluorescence, and electron microscopy on cultures being prepared for flight on the Space Shuttle. For analysis of microfilaments, microtubules, centrosomes, and calcium-requiring events, our standard laboratory protocols had to be modified substantially for experimentation on the Space Shuttle. All manipulations were carried out in a closed culture chamber containing 35 ml artificial sea water as a culture fluid. Unfertilized eggs stored for 24 hours in these chambers were fertilized with sperm diluted in sea water and fixed with concentrated fixatives for final fixation in formaldehyde, taxol, EGTA, and MgCl2(exp -6)H2O for 1 cell to 16 cell stages to preserve cytoskeletal structures for simultaneous analysis with light, immunofluorescence, and electron microscopy, and 1.5 percent glutaraldehyde and 0.4 percent formaldehyde for blastula and plueus stages. The fixed samples wre maintained in chambers without degradation for up to two weeks after which the specimens were processed and analyzed with routine methods. Since complex manipulations are not possible in the closed chambers, the fertilization coat was removed from fixation using 0.5 percent freshly prepared sodium thioglycolate solution at pH 10.0 which provided reliable immunofluorescence staining for microtubules. Sperm/egg fusion, mitosis, cytokinesis, and calcium deposition during spicule formatin in early embryogenesis were found to be without artificial alterations when compared to cells fixed fresh and processed with conventional methods.

Lactate shuttles in nature.

PubMed

Brooks, G A

2002-04-01

Once thought to be the consequence of oxygen lack in contracting skeletal muscle, the glycolytic product lactate is formed and utilized continuously under fully aerobic conditions. "Cell-cell" and "intracellular lactate shuttle" concepts describe the roles of lactate in the delivery of oxidative and gluconeogenic substrates, as well as in cell signalling. Examples of cell-cell shuttles include lactate exchanges between white-glycolytic and red-oxidative fibres within a working muscle bed, between working skeletal muscle and heart, and between tissues of net lactate release and gluconeogenesis. Lactate exchange between astrocytes and neurons that is linked to glutamatergic signalling in the brain is an example of a lactate shuttle supporting cell-cell signalling. Lactate uptake by mitochondria and pyruvate-lactate exchange in peroxisomes are examples of intracellular lactate shuttles. Lactate exchange between sites of production and removal is facilitated by monocarboxylate transport proteins, of which there are several isoforms, and, probably, also by scaffolding proteins. The mitochondrial lactate-pyruvate transporter appears to work in conjunction with mitochondrial lactate dehydrogenase, which permits lactate to be oxidized within actively respiring cells. Hence mitochondria function to establish the concentration and proton gradients necessary for cells with high mitochondrial densities (e.g. cardiocytes) to take up and oxidize lactate. Arteriovenous difference measurements on working cardiac and skeletal muscle beds as well as NMR spectral analyses of these tissues show that lactate is formed and oxidized within the cells of formation in vivo. Glycolysis and lactate oxidation within cells permits high flux rates and the maintenance of redox balance in the cytosol and mitochondria. Other examples of intracellular lactate shuttles include lactate uptake and oxidation in sperm mitochondria and the facilitation of beta-oxidation in peroxisomes by pyruvate

Role of the ARF Tumor Suppressor in Prostate Cancer

DTIC Science & Technology

2006-10-01

also a paucity of rough endoplasmic reticulum compared to wild-type cells ( diamond headed arrows, upper panels) indicating a decrease in protein...cell cycle and cell growth (48). For example, the Tuberous Sclerosis Complex inherited cancer syndrome is caused by germline mutations in either the...J. M. Gardner , F. J. Tavner, D. A. Jans, B. R. Henderson, and T. J. Gonda. 2003. Myb-binding protein 1a is a nucleocytoplasmic shuttling protein that

NASA Contingency Shuttle Crew Support (CSCS) Medical Operations

NASA Technical Reports Server (NTRS)

Adams, Adrien

2010-01-01

The genesis of the space shuttle began in the 1930's when Eugene Sanger came up with the idea of a recyclable rocket plane that could carry a crew of people. The very first Shuttle to enter space was the Shuttle "Columbia" which launched on April 12 of 1981. Not only was "Columbia" the first Shuttle to be launched, but was also the first to utilize solid fuel rockets for U.S. manned flight. The primary objectives given to "Columbia" were to check out the overall Shuttle system, accomplish a safe ascent into orbit, and to return back to earth for a safe landing. Subsequent to its first flight Columbia flew 27 more missions but on February 1st, 2003 after a highly successful 16 day mission, the Columbia, STS-107 mission, ended in tragedy. With all Shuttle flight successes come failures such as the fatal in-flight accident of STS 107. As a result of the STS 107 accident, and other close-calls, the NASA Space Shuttle Program developed contingency procedures for a rescue mission by another Shuttle if an on-orbit repair was not possible. A rescue mission would be considered for a situation where a Shuttle and the crew were not in immediate danger, but, was unable to return to Earth or land safely. For Shuttle missions to the International Space Station (ISS), plans were developed so the Shuttle crew would remain on board ISS for an extended period of time until rescued by a "rescue" Shuttle. The damaged Shuttle would subsequently be de-orbited unmanned. During the period when the ISS Crew and Shuttle crew are on board simultaneously multiple issues would need to be worked including, but not limited to: crew diet, exercise, psychological support, workload, and ground contingency support

Intrepid Space Shuttle Pavilion Opening

NASA Image and Video Library

2012-07-19

The space shuttle Enterprise is seen shortly after the grand opening of the Space Shuttle Pavilion at the Intrepid Sea, Air & Space Museum on Thursday, July 19, 2012 in New York. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Projects

NASA Image and Video Library

1995-06-07

Designed by the mission crew members, the patch for STS-69 symbolizes the multifaceted nature of the flight's mission. The primary payload, the Wake Shield Facility (WSF), is represented in the center by the astronaut emblem against a flat disk. The astronaut emblem also signifies the importance of human beings in space exploration, reflected by the planned space walk to practice for International Space Station (ISS) activities and to evaluate space suit design modifications. The two stylized Space Shuttles highlight the ascent and entry phases of the mission. Along with the two spiral plumes, the stylized Space Shuttles symbolize a NASA first, the deployment and recovery on the same mission of two spacecraft (both the Wake Shield Facility and the Spartan). The constellations Canis Major and Canis Minor represent the astronomy objectives of the Spartan and International Extreme Ultraviolet Hitchhiker (IEH) payload. The two constellations also symbolize the talents and dedication of the support personnel who make Space Shuttle missions possible.

Space Shuttle Projects

NASA Image and Video Library

1995-05-27

The crew patch of STS-72 depicts the Space Shuttle Endeavour and some of the payloads on the flight. The Japanese satellite, Space Flyer Unit (SFU) is shown in a free-flying configuration with the solar array panels deployed. The inner gold border of the patch represents the SFU's distinct octagonal shape. Endeavour’s rendezvous with and retrieval of SFU at an altitude of approximately 250 nautical miles. The Office of Aeronautics and Space Technology's (OAST) flyer satellite is shown just after release from the Remote Manipulator System (RMS). The OAST satellite was deployed at an altitude of 165 nautical miles. The payload bay contains equipment for the secondary payloads - the Shuttle Laser Altimeter (SLA) and the Shuttle Solar Backscatter Ultraviolet Instrument (SSBUV). There were two space walks planned to test hardware for assembly of the International Space Station. The stars represent the hometowns of the crew members in the United States and Japan.

Electromagnetic Compatibility for the Space Shuttle

NASA Technical Reports Server (NTRS)

Scully, Robert C.

2004-01-01

This slide presentation reviews the Space Shuttle electromagnetic compatibility (EMC). It includes an overview of the design of the shuttle with the areas that are of concern for the electromagnetic compatibility. It includes discussion of classical electromagnetic interference (EMI) and the work performed to control the electromagnetic interference. Another area of interest is electrostatic charging and the threat of electrostatic discharge and the attempts to reduce damage to the Shuttle from these possible hazards. The issue of electrical bonding is als reviewed. Lastly the presentation reviews the work performed to protect the shuttle from lightning, both in flight and on the ground.

Planned development of the space shuttle vehicle

NASA Technical Reports Server (NTRS)

1972-01-01

Information pertaining to the planned development of the space shuttle vehicle is presented. The package contains: (1) President's statement; (2) Dr. Fletcher's statement; (3) space shuttle fact sheet; (4) important reasons for the space shuttle.

Space Shuttle Status News Conference

NASA Technical Reports Server (NTRS)

2005-01-01

Richard Gilbech, External Tank "Tiger Team" Lead, begins this space shuttle news conference with detailing the two major objectives of the team. The objectives include: 1) Finding the root cause of the foam loss on STS-114; and 2) Near and long term improvements for the external tank. Wayne Hale, Space Shuttle Program Manager, presents a chart to explain the external tank foam loss during STS-114. He gives a possible launch date for STS-121 after there has been a repair to the foam on the External Tank. He further discusses the changes that need to be made to the surrounding areas of the plant in New Orleans, due to Hurricane Katrina. Bill Gerstemaier, NASA Associate Administrator for Space Operations, elaborates on the testing of the external tank foam loss. The discussion ends with questions from the news media about a fix for the foam, replacement of the tiles, foam loss avoidance, the root cause of foam loss and a possible date for a new external tank to be shipped to NASA Kennedy Space Center.

Space Shuttle Projects

NASA Image and Video Library

1996-03-01

The STS-78 patch links past with present to tell the story of its mission and science through a design imbued with the strength and vitality of the 2-dimensional art of North America's northwest coast Indians. Central to the design is the space Shuttle whose bold lines and curves evoke the Indian image for the eagle, a native American symbol of power and prestige as well as the national symbol of the United States. The wings of the Shuttle suggest the wings of the eagle whose feathers, indicative of peace and friendship in Indian tradition, are captured by the U forms, a characteristic feature of Northwest coast Indian art. The nose of the Shuttle is the strong downward curve of the eagle's beak, and the Shuttle's forward windows, the eagle's eyes, represented through the tapered S forms again typical of this Indian art form. The basic black and red atoms orbiting the mission number recall the original NASA emblem while beneath, utilizing Indian ovoid forms, the major mission scientific experiment package LMS (Life and Materials Sciences) housed in the Shuttle's cargo bay is depicted in a manner reminiscent of totem-pole art. This image of a bird poised for flight, so common to Indian art, is counterpointed by an equally familiar Tsimshian Indian symbol, a pulsating sun with long hyperbolic rays, the symbol of life. Within each of these rays are now encased crystals, the products of this mission's 3 major, high-temperature materials processing furnaces. And as the sky in Indian lore is a lovely open country, home of the Sun Chief and accessible to travelers through a hole in the western horizon, so too, space is a vast and beckoning landscape for explorers launched beyond the horizon. Beneath the Tsimshian sun, the colors of the earth limb are appropriately enclosed by a red border representing life to the Northwest coast Indians. The Indian colors of red, navy blue, white, and black pervade the STS-78 path. To the right of the Shuttle-eagle, the constellation

Tripeptidyl Peptidase II Is Required for c-MYC-Induced Centriole Overduplication and a Novel Therapeutic Target in c-MYC-Associated Neoplasms.

PubMed

Duensing, Stefan; Darr, Sebastian; Cuevas, Rolando; Melquiot, Nadja; Brickner, Anthony G; Duensing, Anette; Münger, Karl

2010-09-01

Centrosome aberrations are frequently detected in c-MYC-associated human malignancies. Here, we show that c-MYC-induced centrosome and centriole overduplication critically depend on the protease tripeptidyl peptidase II (TPPII). We found that TPPII localizes to centrosomes and that overexpression of TPPII, similar to c-MYC, can disrupt centriole duplication control and cause centriole multiplication, a process during which maternal centrioles nucleate the formation of more than a single daughter centriole. We report that inactivation of TPPII using chemical inhibitors or siRNA-mediated protein knockdown effectively reduced c-MYC-induced centriole overduplication. Remarkably, the potent and selective TPPII inhibitor butabindide not only potently suppressed centriole aberrations but also caused significant cell death and growth suppression in aggressive human Burkitt lymphoma cells with c-MYC overexpression. Taken together, these results highlight the role of TPPII in c-MYC-induced centriole overduplication and encourage further studies to explore TPPII as a novel antineoplastic drug target.

Tripeptidyl Peptidase II Is Required for c-MYC–Induced Centriole Overduplication and a Novel Therapeutic Target in c-MYC–Associated Neoplasms

PubMed Central

Duensing, Stefan; Darr, Sebastian; Cuevas, Rolando; Melquiot, Nadja; Brickner, Anthony G.; Duensing, Anette; Münger, Karl

2010-01-01

Centrosome aberrations are frequently detected in c-MYC–associated human malignancies. Here, we show that c-MYC–induced centrosome and centriole overduplication critically depend on the protease tripeptidyl peptidase II (TPPII). We found that TPPII localizes to centrosomes and that overexpression of TPPII, similar to c-MYC, can disrupt centriole duplication control and cause centriole multiplication, a process during which maternal centrioles nucleate the formation of more than a single daughter centriole. We report that inactivation of TPPII using chemical inhibitors or siRNA-mediated protein knockdown effectively reduced c-MYC–induced centriole overduplication. Remarkably, the potent and selective TPPII inhibitor butabindide not only potently suppressed centriole aberrations but also caused significant cell death and growth suppression in aggressive human Burkitt lymphoma cells with c-MYC overexpression. Taken together, these results highlight the role of TPPII in c-MYC–induced centriole overduplication and encourage further studies to explore TPPII as a novel antineoplastic drug target. PMID:21647238

Report of the Space Shuttle Management Independent Review Team

NASA Technical Reports Server (NTRS)

1995-01-01

At the request of the NASA Administrator a team was formed to review the Space Shuttle Program and propose a new management system that could significantly reduce operating costs. Composed of a group of people with broad and extensive experience in spaceflight and related areas, the team received briefings from the NASA organizations and most of the supporting contractors involved in the Shuttle Program. In addition, a number of chief executives from the supporting contractors provided advice and suggestions. The team found that the present management system has functioned reasonably well despite its diffuse structure. The team also determined that the shuttle has become a mature and reliable system, and--in terms of a manned rocket-propelled space launch system--is about as safe as today's technology will provide. In addition, NASA has reduced shuttle operating costs by about 25 percent over the past 3 years. The program, however, remains in a quasi-development mode and yearly costs remain higher than required. Given the current NASA-contractor structure and incentives, it is difficult to establish cost reduction as a primary goal and implement changes to achieve efficiencies. As a result, the team sought to create a management structure and associated environment that enables and motivates the Program to further reduce operational costs. Accordingly, the review team concluded that the NASA Space Shuttle Program should (1) establish a clear set of program goals, placing a greater emphasis on cost-efficient operations and user-friendly payload integration; (2) redefine the management structure, separating development and operations and disengaging NASA from the daily operation of the space shuttle; and (3) provide the necessary environment and conditions within the program to pursue these goals.

Observations of thermal ion influxes about the space shuttle

NASA Technical Reports Server (NTRS)

Grebowsky, Joe M.; Schaefer, A.

1990-01-01

Ion mass spectrometer measurements made as part of the University of Iowa's Plasma Diagnostic Package on the STS-3 and Spacelab 2 Space Shuttle missions sampled a variety of ion composition and collected ion current responses to gas emissions from the vehicle. The only other shuttle ion measurements were made by an Air Force Geophysics Laboratory (AFGL) quadrupole spectrometer flown on STS-4. Gas emissions change the distribution of the incoming plasma through scattering and charge transfer processes. A background flux of contaminant ion species (mostly relating to water) always exists in the near vicinity of the shuttle with a magnitude which is dependent on the look direction of the spectrometer but which varies differently with changes in the angle of attack than that of the ambient ions. There is a near shuttle wake cavity in the contaminant ion distributions which has a different spatial configuration than the wake of the ambient ions. Although water dumps produce the most persistent ion perturbations, the sources for ion current modification were best delineated from measurements made when only one or two of the Reaction Control System thrusters fired for a relatively long duration. Contaminant ion perturbations associated with such firings were observed to persist for the order of a second after the cessation of the firings. The dense thruster plumes are efficient collisional, charge exchange barriers to the passage of ambient ions. Collected ion current perturbations were more evident for firings of the rear verniers, whose plumes scatter off projecting surfaces, than for the nose thrusters. The effect of the Vernier firings was found to depend not only on the location and attitude of the spectrometer with respect to the shuttle and thruster plume direction, but also on the orientation of the local magnetic field with respect to the shuttle velocity.

Report of the Space Shuttle Management Independent Review Team

NASA Astrophysics Data System (ADS)

1995-02-01

At the request of the NASA Administrator a team was formed to review the Space Shuttle Program and propose a new management system that could significantly reduce operating costs. Composed of a group of people with broad and extensive experience in spaceflight and related areas, the team received briefings from the NASA organizations and most of the supporting contractors involved in the Shuttle Program. In addition, a number of chief executives from the supporting contractors provided advice and suggestions. The team found that the present management system has functioned reasonably well despite its diffuse structure. The team also determined that the shuttle has become a mature and reliable system, and--in terms of a manned rocket-propelled space launch system--is about as safe as today's technology will provide. In addition, NASA has reduced shuttle operating costs by about 25 percent over the past 3 years. The program, however, remains in a quasi-development mode and yearly costs remain higher than required. Given the current NASA-contractor structure and incentives, it is difficult to establish cost reduction as a primary goal and implement changes to achieve efficiencies. As a result, the team sought to create a management structure and associated environment that enables and motivates the Program to further reduce operational costs. Accordingly, the review team concluded that the NASA Space Shuttle Program should (1) establish a clear set of program goals, placing a greater emphasis on cost-efficient operations and user-friendly payload integration; (2) redefine the management structure, separating development and operations and disengaging NASA from the daily operation of the space shuttle; and (3) provide the necessary environment and conditions within the program to pursue these goals.

Space Shuttle Project

NASA Image and Video Library

1995-10-20

A Great Blue Heron seems oblivious to the tremendous spectacle of light and sound generated by a Shuttle liftoff, as the Space Shuttle Columbia (STS-73) soars skyward from Launch Pad 39B. Columbia's seven member crew's mission included continuing experimentation in the Marshall managed payloads including the United States Microgravity Laboratory 2 (USML-2) and the keel-mounted accelerometer that characterizes the very low frequency acceleration environment of the orbiter payload bay during space flight, known as the Orbital Acceleration Research Experiment (OARE).

Using sea urchin gametes and zygotes to investigate centrosome duplication.

PubMed

Sluder, Greenfield

2016-01-01

Centriole structure and function in the sea urchin zygote parallel those in mammalian somatic cells. Here, I briefly introduce the properties and attributes of the sea urchin system that make it an attractive platform for the study of centrosome and centriole duplication. These attributes apply to all echinoderms readily available from commercial suppliers: sea urchins, sand dollars, and starfish. I list some of the practical aspects of the system that make it a cost- and time-effective system for experimental work and then list properties that are a "tool kit" that can be used to conduct studies that would not be practical, or in some cases not possible, with mammalian somatic cells. Since centrioles organize and localize the pericentriolar material that nucleates the astral arrays of microtubules (Bobinnec et al. in J Cell Biol 143(6):1575-1589, 1998), the pattern of aster duplication over several cell cycles can be used as a reliable measure for centriole duplication (Sluder and Rieder in J Cell Biol 100(3):887-896, 1985). Descriptions of the methods my laboratory has used to handle and image echinoderm zygotes are reviewed in Sluder et al. (Methods Cell Biol 61:439-472, 1999). Also included is a bibliography of papers that describe additional methods.

Embryonic expression of shuttle craft, a Drosophila gene involved in neuron development, is associated with adult lifespan.

PubMed

Roshina, Natalia V; Symonenko, Alexander V; Krementsova, Anna V; Trostnikov, Mikhail V; Pasyukova, Elena G

2014-12-01

Despite the progress in aging research that highlights the role of the nervous system in longevity, whether genes that control development and consequently structure of the nervous system affect lifespan is unclear. We demonstrated that a mutation inshuttle craft, a gene involved in the nervous system development, increased the lifespan of unmated females and decreased the lifespan of mated females, without affecting males. Precise reversions of the mutation lead to the restoration of the lifespan specific to control females. In mutant unmated females, increased lifespan was associated with elevated locomotion at older ages, indicating slowed aging. In mutant mated females, reproduction was decreased compared to controls, indicating a lack of tradeoff between this trait and lifespan. No differences in shuttle craft transcription were observed between whole bodies, ovaries, and brains of mutant and control females of different ages, either unmated or mated. The amount of shuttle craft transcript appeared to be substantially decreased in mutant embryos. Our results demonstrated that a gene that regulates development of the nervous system might also influence longevity, and thus expanded the spectrum of genes involved in lifespan control. We hypothesize that this "carry-over" effect might be the result of transcription regulation in embryos.

Space Shuttle Discovery Fly-By

NASA Image and Video Library

2012-04-17

Space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) flies over the Steven F. Udvar-Hazy Center, Tuesday, April 17, 2012, in Washington. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Smithsonian Institution/Eric Long)

EA Shuttle Document Retention Effort

NASA Technical Reports Server (NTRS)

Wagner, Howard A.

2010-01-01

This slide presentation reviews the effort of code EA at Johnson Space Center (JSC) to identify and acquire databases and documents from the space shuttle program that are adjudged important for retention after the retirement of the space shuttle.

Space Shuttle Placement Announcement

NASA Image and Video Library

2011-04-12

Pilot of the first space shuttle mission, STS-1, Bob Crippen speaks at an event where NASA Administrator Charles Bolden announced where four space shuttle orbiters will be permanently displayed at the conclusion of the Space Shuttle Program, Tuesday, April 12, 2011, at Kennedy Space Center in Cape Canaveral, Fla. The four orbiters, Enterprise, which currently is on display at the Smithsonian's Steven F. Udvar-Hazy Center near Washington Dulles International Airport, will move to the Intrepid Sea, Air & Space Museum in New York, Discovery will move to Udvar-Hazy, Endeavour will be displayed at the California Science Center in Los Angeles and Atlantis, in background, will be displayed at the Kennedy Space Center Visitor’s Complex. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Placement Announcement

NASA Image and Video Library

2011-04-12

The space shuttle Atlantis is seen in the Orbiter Processing Facility at an event where NASA Administrator Charles Bolden announced where four space shuttle orbiters will be permanently displayed at the conclusion of the Space Shuttle Program, Tuesday, April 12, 2011, at Kennedy Space Center in Cape Canaveral, Fla. The four orbiters, Enterprise, which currently is on display at the Smithsonian's Steven F. Udvar-Hazy Center near Washington Dulles International Airport, will move to the Intrepid Sea, Air & Space Museum in New York, Discovery will move to Udvar-Hazy, Endeavour will be displayed at the California Science Center in Los Angeles and Atlantis, will be displayed at the Kennedy Space Center Visitor’s Complex. Photo Credit: (NASA/Bill Ingalls)

Space Shuttle Placement Announcement

NASA Image and Video Library

2011-04-12

Pilot of the first space shuttle mission, STS-1, Bob Crippen speaks at an event where NASA Administrator Charles Bolden announced where four space shuttle orbiters will be permanently displayed at the conclusion of the Space Shuttle Program, Tuesday, April 12, 2011, at Kennedy Space Center in Cape Canaveral, Fla. The four orbiters, Enterprise, which currently is on display at the Smithsonian's Steven F. Udvar-Hazy Center near Washington Dulles International Airport, will move to the Intrepid Sea, Air & Space Museum in New York, Discovery will move to Udvar-Hazy, Endeavour will be displayed at the California Science Center in Los Angeles and Atlantis, in background, will be displayed at the Kennedy Space Center Visitor’s Complex. Photo Credit: (NASA/Bill Ingalls)

Experiment definition phase shuttle laboratory (LDRL-10.6 experiment): Shuttle sortie to elliptical orbit satellite

NASA Technical Reports Server (NTRS)

Goodwin, F. E.; Nussmeier, T. A.; Stokes, L. S.; Vourgourakis, E. J.

1976-01-01

The following topics were reviewed: (1) design options for shuttle terminal, (2) elliptical orbit satellite design options, (3) shuttle terminal details, (4) technology status and development requirements, (5) transmitter technology, and (6) carbon dioxide laser life studies.

Intrepid Space Shuttle Pavilion Opening

NASA Image and Video Library

2012-07-19

Former NASA Astronaut and Enterprise Commander Joe Engle looks at an exhibit in the Intrepid Sea, Air & Space Museum's Space Shuttle Pavilion where the space shuttle Enterprise is on Thursday, July 19, 2012 in New York. Photo Credit: (NASA/Bill Ingalls)

STS-38 Space Shuttle mission report

NASA Technical Reports Server (NTRS)

Camp, David W.; Germany, D. M.; Nicholson, Leonard S.

1991-01-01

The STS-38 Space Shuttle Program Mission Report contains a summary of the vehicle subsystem activities on this thirty-seventh flight of the Space Shuttle and the seventh flight of the Orbiter vehicle Atlantis (OV-104). In addition to the Atlantis vehicle, the flight vehicle consisted of an External Tank (ET) (designated as ET-40/LWT-33), three Space Shuttle main engines (SSME's) (serial numbers 2019, 2022, 2027), and two Solid Rocket Boosters (SRB's), designated as BI-039. The STS-38 mission was a classified Department of Defense mission, and as much, the classified portions of the mission are not presented in this report. The sequence of events for this mission is shown. The significant problems that occurred in the Space Shuttle Orbiter subsystem during the mission are summarized and the official problem tracking list is presented. In addition, each Space Shuttle Orbiter problem is cited in the subsystem discussion.

Success Legacy of the Space Shuttle Program: Changes in Shuttle Post Challenger and Columbia

NASA Technical Reports Server (NTRS)

Jarrell, George

2010-01-01

This slide presentation reviews the legacy of successes in the space shuttle program particularly with regards to the changes in the culture of NASA's organization after the Challenger and Columbia accidents and some of the changes to the shuttles that were made manifest as a result of the accidents..

Space shuttle on-orbit flight control software requirements, preliminary version

NASA Technical Reports Server (NTRS)

1975-01-01

Software modules associated with various flight control functions for the space shuttle orbiter are described. Data flow, interface requirements, initialization requirements and module sequencing requirements are considered. Block diagrams and tables are included.

Stroke Induces Nuclear Shuttling of Histone Deacetylase 4.

PubMed

Kassis, Haifa; Shehadah, Amjad; Chopp, Michael; Roberts, Cynthia; Zhang, Zheng Gang

2015-07-01

Histone deacetylases (HDACs) 4 and 5 are abundantly expressed in the brain and have been implicated in the regulation of neurodegeneration. Under physiological conditions, HDACs 4 and 5 are expressed in the cytoplasm of brain cells where they cannot directly access chromatin. In response to external stimuli, they can shuttle to the nucleus and regulate gene expression. However, the effect of stroke on nuclear shuttling of HDACs 4 and 5 remains unknown. Using a rat model of middle cerebral artery occlusion, we examined the subcellular localization of HDACs 4 and 5 in the peri-infarct cortex during brain repair after stroke. Stroke significantly increased nuclear HDAC4 immunoreactivity in neurons, but not in astrocytes or in oligodendrocytes, of the peri-infarct cortex at 2, 7, and 14 days after middle cerebral artery occlusion. Neurons with nuclear HDAC4 immunoreactivity distributed across all layers of the peri-infarct cortex and were Ctip2+ excitatory and parvalbumin+ inhibitory neurons. These neurons were not TUNEL or BrdU positive. Furthermore, nuclear HDAC4 immunoreactivity was positively and significantly correlated with increased dendritic, axonal, and myelin densities as determined by microtubule-associated protein 2, phosphorylated neurofilament heavy chain, and myelin basic protein, respectively. Unlike HDAC4, stroke did not alter nuclear localization of HDAC5. Our data show that stroke induces nuclear shuttling of HDAC4 in neurons in the peri-infarct cortex, and that increased nuclear HDAC4 is strongly associated with neuronal remodeling but not with neuronal cell death, suggesting a role for nuclear HDAC4 in promoting neuronal recovery after ischemic injury. © 2015 American Heart Association, Inc.

Evaluation philosophy for shuttle launched payloads

NASA Technical Reports Server (NTRS)

Heuser, R. E.

1975-01-01

Some approaches to space-shuttle payload evaluation are examined. Issues considered include subsystem replacement in low-cost modular spacecraft (LCMS), validation of spacelab payloads, the use of standard components in shuttle-era spacecraft, effects of shuttle-induced environments on payloads, and crew safety. The LCMS is described, and goals are discussed for its evaluation program. Concepts regarding how the evaluation should proceed are considered.

STS-80 Space Shuttle Mission Report

NASA Technical Reports Server (NTRS)

Fricke, Robert W., Jr.

1997-01-01

The STS-80 Space Shuttle Program Mission Report summarizes the Payload activities as well as the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Reusable Solid Rocket Motor (RSRM), and the Space Shuttle main engine (SSME) systems performance during the eightieth flight of the Space Shuttle Program, the fifty-fifth flight since the return-to-flight, and the twenty-first flight of the Orbiter Columbia (OV-102).

Space Shuttle Projects

NASA Image and Video Library

1990-10-06

Launched aboard the Space Shuttle Discovery on October 6, 1990 at 7:47:15 am (EDT), the STS-41 mission consisted of 5 crew members. Included were Richard N. Richards, commander; Robert D. Cabana, pilot; and Bruce E. Melnick, Thomas D. Akers, and William M. Shepherd, all mission specialists. The primary payload for the mission was the European Space Agency (ESA) built Ulysses Space Craft made to explore the polar regions of the Sun. Other main payloads and experiments included the Shuttle Solar Backscatter Ultraviolet (SSBUV) experiment and the INTELSAT Solar Array Coupon (ISAC).

Shuttle mission plans

NASA Technical Reports Server (NTRS)

Visentine, J. T.; Lee, C. M.

1978-01-01

Shuttle mission plans recently developed by NASA for the time period 1980-1991 are presented. Standard and optional services, which will be available to users of the Space Transportation System (STS) when it becomes operational in the 1980's, are described. Pricing policies established by NASA to encourage use of the STS by commercial, foreign and other U.S. Government users are explained. The small Self-Contained Payload Program, which will make space flight opportunities available to private citizens and individual experimenters who wish to use the Space Shuttle for investigative research, is discussed.

Space Shuttle Project

NASA Image and Video Library

1993-04-08

The second try works like a charm as the Space Shuttle Discovery (STS-56) lifts off from Launch Pad 39B. The first attempt to launch was halted at T-11 seconds on April 6th. Aboard for the second shuttle mission of 1993 were a crew of five and the Atmospheric Laboratory for Applications and Science 2 (ATLAS 2), the second in a series of missions to study the sun's energy output and Earth's middle atmosphere chemical make-up, and how these factors affect levels of ozone.

Space Shuttle Project

NASA Image and Video Library

1992-01-22

Onboard Space Shuttle Discovery (STS-42) the seven crewmembers pose for a traditional in-space portrait in the shirt-sleeve environment of the International Microgravity Laboratory (IML-1) science module in the Shuttle's cargo bay. Pictured are (clockwise from top),Commander Ronald J. Grabe, payload commander Norman E. Thagard, payload specialist Roberta L. Bondar; mission specialists William F. Readdy and David C. Hilmers; pilot Stephen S. Oswald and payload specialist Ulf Merbold. The rotating chair, used often in biomedical tests on the eight-day flight, is in center frame.

STS-92 - Towing of Shuttle Discovery and Boeing 747 Shuttle Carrier Aircraft (SCA)

NASA Image and Video Library

2000-11-02

The Space Shuttle Discovery sits atop one of NASA’s modified Boeing 747 Shuttle Carrier Aircraft as the unusual piggyback duo is towed along a taxiway at NASA’s Dryden Flight Research Center at Edwards, California. The Discovery was ferried from NASA Dryden to NASA’s Kennedy Space Center in Florida on November 2, 2000, after extensive pre-ferry servicing and preparations.

Space Shuttle Projects Overview to Columbia Air Forces War College

NASA Technical Reports Server (NTRS)

Singer, Jody; McCool, Alex (Technical Monitor)

2000-01-01

This paper presents, in viewgraph form, a general overview of space shuttle projects. Some of the topics include: 1) Space Shuttle Projects; 2) Marshall Space Flight Center Space Shuttle Projects Office; 3) Space Shuttle Propulsion systems; 4) Space Shuttle Program Major Sites; 5) NASA Office of Space flight (OSF) Center Roles in Space Shuttle Program; 6) Space Shuttle Hardware Flow; and 7) Shuttle Flights To Date.

Microbiological Lessons Learned from the Space Shuttle

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Ott, C. Mark; Bruce, Rebekah; Castro, Victoria A.; Mehta, Satish K.

2011-01-01

After 30 years of being the centerpiece of NASA s human spacecraft, the Space Shuttle will retire. This highly successful program provided many valuable lessons for the International Space Station (ISS) and future spacecraft. Major microbiological risks to crewmembers include food, water, air, surfaces, payloads, animals, other crewmembers, and ground support personnel. Adverse effects of microorganisms are varied and can jeopardize crew health and safety, spacecraft systems, and mission objectives. Engineering practices and operational procedures can minimize the negative effects of microorganisms. To minimize problems associated with microorganisms, appropriate steps must begin in the design phase of new spacecraft or space habitats. Spacecraft design must include requirements to control accumulation of water including humidity, leaks, and condensate on surfaces. Materials used in habitable volumes must not contribute to microbial growth. Use of appropriate materials and the implementation of robust housekeeping that utilizes periodic cleaning and disinfection will prevent high levels of microbial growth on surfaces. Air filtration can ensure low levels of bioaerosols and particulates in the breathing air. The use of physical and chemical steps to disinfect drinking water coupled with filtration can provide safe drinking water. Thorough preflight examination of flight crews, consumables, and the environment can greatly reduce pathogens in spacecraft. The advances in knowledge of living and working onboard the Space Shuttle formed the foundation for environmental microbiology requirements and operations for the International Space Station (ISS) and future spacecraft. Research conducted during the Space Shuttle Program resulted in an improved understanding of the effects of spaceflight on human physiology, microbial properties, and specifically the host-microbe interactions. Host-microbe interactions are substantially affected by spaceflight. Astronaut immune

Shuttle Discovery Landing at Palmdale, California, Maintenance Facility

NASA Technical Reports Server (NTRS)

1995-01-01

NASA Dryden Flight Research Center pilot Tom McMurtry lands NASA's Shuttle Carrier Aircraft with Space Shuttle Discovery attached at Rockwell Aerospace's Palmdale, California, facility about 1:00 p.m. Pacific Daylight Time (PDT). There for nine months of scheduled maintenance, Discovery and the 747 were completing a two-day flight from Kennedy Space Center, Florida, that began at 7:04 a.m. Eastern Standard Time on 27 September and included an overnight stop at Salt Lake City International Airport, Utah. At the conclusion of this mission, Discovery had flown 21 shuttle missions, totaling more than 142 days in orbit. Space Shuttles are the main element of America's Space Transportation System and are used for space research and other space applications. The shuttles are the first vehicles capable of being launched into space and returning to Earth on a routine basis. Space Shuttles are used as orbiting laboratories in which scientists and mission specialists conduct a wide variety of scientific experiments. Crews aboard shuttles place satellites in orbit, rendezvous with satellites to carry out repair missions and return them to space, and retrieve satellites and return them to Earth for refurbishment and reuse. Space Shuttles are true aerospace vehicles. They leave Earth and its atmosphere under rocket power provided by three liquid-propellant main engines with two solid-propellant boosters attached plus an external liquid-fuel tank. After their orbital missions, they streak back through the atmosphere and land like airplanes. The returning shuttles, however, land like gliders, without power and on runways. Other rockets can place heavy payloads into orbit, but, they can only be used once. Space Shuttles are designed to be continually reused. When Space Shuttles are used to transport complete scientific laboratories into space, the laboratories remain inside the payload bay throughout the mission. They are then removed after the Space Shuttle returns to Earth and

Space Shuttle ice nuclei

NASA Astrophysics Data System (ADS)

Turco, R. P.; Toon, O. B.; Whitten, R. C.; Cicerone, R. J.

1982-08-01

Estimates are made showing that, as a consequence of rocket activity in the earth's upper atmosphere in the Shuttle era, average ice nuclei concentrations in the upper atmosphere could increase by a factor of two, and that an aluminum dust layer weighing up to 1000 tons might eventually form in the lower atmosphere. The concentrations of Space Shuttle ice nuclei (SSIN) in the upper troposphere and lower stratosphere were estimated by taking into account the composition of the particles, the extent of surface poisoning, and the size of the particles. Calculated stratospheric size distributions at 20 km with Space Shuttle particulate injection, calculated SSIN concentrations at 10 and 20 km altitude corresponding to different water vapor/ice supersaturations, and predicted SSIN concentrations in the lower stratosphere and upper troposphere are shown.

Space Shuttle Discovery Fly-Over

NASA Image and Video Library

2012-04-17

Spectators watch as space shuttle Discovery, mounted atop a NASA 747 Shuttle Carrier Aircraft (SCA) flies over the National Air and Space Museum’s Steven F. Udvar-Hazy Center, Tuesday, April 17, 2012, in Chantilly, Va. Discovery, the first orbiter retired from NASA’s shuttle fleet, completed 39 missions, spent 365 days in space, orbited the Earth 5,830 times, and traveled 148,221,675 miles. NASA will transfer Discovery to the National Air and Space Museum to begin its new mission to commemorate past achievements in space and to educate and inspire future generations of explorers. Photo Credit: (NASA/Carla Cioffi)

Progesterone After Estradiol Modulates Shuttle-Cage Escape by Facilitating Volition

PubMed Central

Mayeaux, Darryl J.; Tandle, Sarah M.; Cilano, Sean M.; Fitzharris, Matthew J.

2015-01-01

In animal models of depression, depression is defined as performance on a learning task. That task is typically escaping a mild electric shock in a shuttle cage by moving from one side of the cage to the other. Ovarian hormones influence learning in other kinds of tasks, and these hormones are associated with depressive symptoms in humans. The role of these hormones in shuttle-cage escape learning, however, is less clear. This study manipulated estradiol and progesterone in ovariectomized female rats to examine their performance in shuttle-cage escape learning without intentionally inducing a depressive-like state. Progesterone, not estradiol, within four hours of testing affected latencies to escape. The improvement produced by progesterone was in the decision to act, not in the speed of learning or speed of escaping. This parallels depression in humans in that depressed people are slower in volition, in their decisions to take action. PMID:26823653

Shuttle Hitchhiker Experiment Launcher System (SHELS)

NASA Technical Reports Server (NTRS)

Daelemans, Gerry

1999-01-01

NASA's Goddard Space Flight Center Shuttle Small Payloads Project (SSPP), in partnership with the United States Air Force and NASA's Explorer Program, is developing a Shuttle based launch system called SHELS (Shuttle Hitchhiker Experiment Launcher System), which shall be capable of launching up to a 400 pound spacecraft from the Shuttle cargo bay. SHELS consists of a Marman band clamp push-plate ejection system mounted to a launch structure; the launch structure is mounted to one Orbiter sidewall adapter beam. Avionics mounted to the adapter beam will interface with Orbiter electrical services and provide optional umbilical services and ejection circuitry. SHELS provides an array of manifesting possibilities to a wide range of satellites.

The Representative Shuttle Environmental Control System

NASA Technical Reports Server (NTRS)

Brose, H. F.; Greenwood, F. H.; Thompson, C. D.; Willis, N. C.

1974-01-01

The Representative Shuttle Environmental Control System (RSECS) program was conceived to provide NASA with a prototype system representative of the Shuttle Environmental Control System (ECS). Discussed are the RSECS program objectives, predicated on updating and adding to the early system as required to retain its usefulness during the Shuttle ECS development and qualification effort. Ultimately, RSECS will be replaced with a flight-designed system using either refurbished development or qualification equipment to provide NASA with a flight simulation capability during the Shuttle missions. The RSECS air revitalization subsystem and the waste management support subsystem are being tested. A water coolant subsystem and a freon coolant subsystem are in the development and planning phases.

Space Shuttle Projects

NASA Image and Video Library

1993-05-01

Designed by members of the flight crew, the STS-58 insignia depicts the Space Shuttle Columbia with a Spacelab module in its payload bay in orbit around Earth. The Spacelab and the lettering Spacelab Life Sciences ll highlight the primary mission of the second Space Shuttle flight dedicated to life sciences research. An Extended Duration Orbiter (EDO) support pallet is shown in the aft payload bay, stressing the scheduled two-week duration of the longest Space Shuttle mission to date. The hexagonal shape of the patch depicts the carbon ring, a molecule common to all living organisms. Encircling the inner border of the patch is the double helix of DNA, representing the genetic basis of life. Its yellow background represents the sun, energy source for all life on Earth. Both medical and veterinary caducei are shown to represent the STS- 58 life sciences experiments. The position of the spacecraft in orbit about Earth with the United States in the background symbolizes the ongoing support of the American people for scientific research intended to benefit all mankind.

NASA's modified Boeing 747 Shuttle Carrier Aircraft with the Space Shuttle Endeavour on top lifts of

NASA Technical Reports Server (NTRS)

2001-01-01

NASA's modified Boeing 747 Shuttle Carrier Aircraft with the Space Shuttle Endeavour on top lifts off from Edwards Air Force Base to begin its ferry flight back to the Kennedy Space Center in Florida.

Space shuttle propulsion systems

NASA Technical Reports Server (NTRS)

Bardos, Russell

1991-01-01

This is a presentation of view graphs. The design parameters are given for the redesigned solid rocket motor (RSRM), the Advanced Solid Rocket Motor (ASRM), Space Shuttle Main Engine (SSME), Solid Rocket Booster (SRB) separation motor, Orbit Maneuvering System (OMS), and the Reaction Control System (RCS) primary and Vernier thrusters. Space shuttle propulsion issues are outlined along with ASA program definition, ASA program selection methodology, its priorities, candidates, and categories.

Upgrading the Space Shuttle.

DTIC Science & Technology

1999-01-01

Motors, Honda , Toyota , and Nissan ). By learning from and applying the technologies developed elsewhere, NASA could greatly leverage its funding for...assessing risks to the shuttle. The committee believes that this tool has the potential to be very helpful in assessing and comparing the impact of...environmental regulations). Figure 2-2 shows how the S&PU budget compared to the total shuttle budget during four different years since 1985

Space Shuttle aerothermodynamic data report, phase C