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1

Phosphoinositide 3-kinase-dependent Ras activation by tauroursodesoxycholate in rat liver.  

PubMed

Ursodesoxycholic acid, widely used for the treatment of cholestatic liver disease, causes choleretic, anti-apoptotic and immunomodulatory effects. Here the effects on choleresis of its taurine conjugate tauroursodesoxycholate (TUDC), which is present in the enterohepatic circulation, were correlated with the activation of important elements of intracellular signal transduction in cultured rat hepatocytes and perfused rat liver. TUDC induced a time- and concentration-dependent activation of the small GTP-binding protein Ras and of phosphoinositide 3-kinase (PI 3-kinase) in cultured hepatocytes. Ras activation was dependent on PI 3-kinase activity, without the involvement of protein kinase C- and genistein-sensitive tyrosine kinases. Ras activation by TUDC was followed by an activation of the mitogen-activated protein kinases extracellular-signal-regulated kinase-1 (Erk-1) and Erk-2. In perfused rat liver, PI 3-kinase inhibitors largely abolished the stimulatory effect of TUDC on taurocholate excretion, suggesting an important role for a PI 3-kinase/Ras/Erk pathway in the choleretic effect of TUDC. PMID:10926845

Kurz, A K; Block, C; Graf, D; Dahl, S V; Schliess, F; Häussinger, D

2000-08-15

2

Aromatase induction in tamoxifen-resistant breast cancer: Role of phosphoinositide 3-kinase-dependent CREB activation.  

PubMed

Estrogens are important for the development and growth of estrogen receptor (ER)-positive breast cancer, for which anti-estrogen therapy is one of the most effective treatments. However, its efficacy can be limited by either de novo or acquired resistance. Aromatase is a key enzyme for the biosynthesis of estrogens, and inhibition of this enzyme leads to profound hypoestrogenism. Here, we found that the basal expression and activity of aromatase were significantly increased in tamoxifen (TAM)-resistant human breast cancer (TAMR-MCF-7) cells compared to control MCF-7 cells. We further revealed that aromatase immunoreactivity in tumor tissues was increased in recurrence group after TAM therapy compared to non-recurrence group after TAM therapy. Phosphorylation of Akt, extracellular signal-regulated kinase (ERK), and p38 kinase were all increased in TAMR-MCF-7 cells. Inhibition of phosphoinositide 3-kinase (PI3K) suppressed the transactivation of the aromatase gene and its enzyme activity. Furthermore, we have also shown that PI3K/Akt-dependent cAMP-response element binding protein (CREB) activation was required for the enhanced expression of aromatase in TAMR-MCF-7 cells. Our findings suggest that aromatase expression is up-regulated in TAM-resistant breast cancer via PI3K/Akt-dependent CREB activation. PMID:24836190

Phuong, Nguyen Thi Thuy; Lim, Sung Chul; Kim, Young Mi; Kang, Keon Wook

2014-08-28

3

Apelin-13 Inhibits Large-Conductance Ca2+-Activated K+ Channels in Cerebral Artery Smooth Muscle Cells via a PI3-Kinase Dependent Mechanism  

PubMed Central

Apelin-13 causes vasoconstriction by acting directly on APJ receptors in vascular smooth muscle (VSM) cells; however, the ionic mechanisms underlying this action at the cellular level remain unclear. Large-conductance Ca2+-activated K+ (BKCa) channels in VSM cells are critical regulators of membrane potential and vascular tone. In the present study, we examined the effect of apelin-13 on BKCa channel activity in VSM cells, freshly isolated from rat middle cerebral arteries. In whole-cell patch clamp mode, apelin-13 (0.001-1 ?M) caused concentration-dependent inhibition of BKCa in VSM cells. Apelin-13 (0.1 µM) significantly decreased BKCa current density from 71.25±8.14 pA/pF to 44.52±7.10 pA/pF (n=14 cells, P<0.05). This inhibitory effect of apelin-13 was confirmed by single channel recording in cell-attached patches, in which extracellular application of apelin-13 (0.1 µM) decreased the open-state probability (NPo) of BKCa channels in freshly isolated VSM cells. However, in inside-out patches, extracellular application of apelin-13 (0.1µM) did not alter the NPo of BKCa channels, suggesting that the inhibitory effect of apelin-13 on BKCa is not mediated by a direct action on BKCa. In whole cell patches, pretreatment of VSM cells with LY-294002, a PI3-kinase inhibitor, markedly attenuated the apelin-13-induced decrease in BKCa current density. In addition, treatment of arteries with apelin-13 (0.1 µM) significantly increased the ratio of phosphorylated-Akt/total Akt, indicating that apelin-13 significantly increases PI3-kinase activity. Taken together, the data suggest that apelin-13 inhibits BKCa channel via a PI3-kinase-dependent signaling pathway in cerebral artery VSM cells, which may contribute to its regulatory action in the control of vascular tone. PMID:24386141

O'Rourke, Stephen T.; Sun, Chengwen

2013-01-01

4

Cellular Notch responsiveness is defined by phosphoinositide 3-kinase-dependent signals  

PubMed Central

Background Notch plays a wide-ranging role in controlling cell fate, differentiation and development. The PI3K-Akt pathway is a similarly conserved signalling pathway which regulates processes such as differentiation, proliferation and survival. Mice with disrupted Notch and PI3K signalling show phenotypic similarities during haematopoietic cell development, suggesting functional interaction between these pathways. Results We show that cellular responsiveness to Notch signals depends on the activity of the PI3K-Akt pathway in cells as diverse as CHO cells, primary T-cells and hippocampal neurons. Induction of the endogenous PI3K-Akt pathway in CHO cells (by the insulin pathway), in T-cells (via TCR activation) or in neurons (via TrKB activation) potentiates Notch-dependent responses. We propose that the PI3K-Akt pathway exerts its influence on Notch primarily via inhibition of GSK3-beta, a kinase known to phosphorylate and regulate Notch signals. Conclusion The PI3K-Akt pathway acts as a "gain control" for Notch signal responses. Since physiological levels of intracellular Notch are often low, coincidence with PI3K-activation may be crucial for induction of Notch-dependent responses. PMID:16507111

Mckenzie, Grahame; Ward, George; Stallwood, Yvette; Briend, Emmanuel; Papadia, Sofia; Lennard, Andrew; Turner, Martin; Champion, Brian; Hardingham, Giles E

2006-01-01

5

Platelet-Derived Growth Factor Reorganizes the Actin Cytoskeleton through 3-Phosphoinositide-Dependent and 3-Phosphoinositide-Independent Mechanisms in Human Mesangial Cells  

Microsoft Academic Search

Background: Platelet-derived growth factor (PDGF) is a potent activator of mesangial cell proliferation and migration. Although phosphoinositide 3-kinase (PI3K) enzymes are important downstream targets of the PDGF receptor, the contribution made by their 3-phosphoinositide products in the reorganization of actin cytoskeleton and focal adhesions has been questioned. Methods and Results: Pharmacological inhibition of the PI3K activity blocks PDGF-induced migration of

Lisa Harper; Yasuyo Kashiwagi; Charles D. Pusey; Bruce M. Hendry; Jan Domin

2007-01-01

6

Small GTPases and phosphoinositides in the regulatory mechanisms of macropinosome formation and maturation  

PubMed Central

Macropinosome formation requires the sequential activation of numerous signaling pathways that coordinate the actin-driven formation of plasma membrane protrusions (ruffles) and circular ruffles (macropinocytic cups), followed by the closure of these macropinocytic cups into macropinosomes. In the process of macropinosome formation, localized productions of phosphoinositides such as PI(4,5)P2 and PI(3,4,5)P3 spatiotemporally orchestrate actin polymerization and rearrangement through recruiting and activating a variety of actin-associated proteins. In addition, the sequential activation of small GTPases, which are known to be master regulators of the actin cytoskeleton, plays a pivotal role in parallel with phosphoinositides. To complete macropinosome formation, phosphoinositide breakdown and Rho GTPase deactivation must occur in appropriate timings. After the nascent macropinosomes are formed, phosphoinositides and several Rab GTPases control macropinosome maturation by regulating vesicle trafficking and membrane fusion. In this review, we summarize recent advances in our understanding of the critical functions of phosphoinositide metabolism and small GTPases in association with their downstream effectors in macropinocytosis.

Egami, Youhei; Taguchi, Tomohiko; Maekawa, Masashi; Arai, Hiroyuki; Araki, Nobukazu

2014-01-01

7

Structural Mapping of the Catalytic Mechanism for a Mammalian Phosphoinositide-Specific Phospholipase C,  

E-print Network

.30-2.95 Ã? resolution. The inositol phosphates used in this study mimic the binding of substrates of phosphoinositide-specific phospholipase C-1 from rat with calcium and inositol phosphates have been determined at 2 phosphates. The structures are consistent with bidentate liganding of the catalytic calcium to the inositol

Williams, Roger L.

8

Bilobalide regulates soluble amyloid precursor protein release via phosphatidyl inositol 3 kinase-dependent pathway  

Microsoft Academic Search

Bilobalide (BB) is a sesquiterpenoid extracted from Ginkgo biloba leaves. An increasing number of studies have demonstrated its neuroprotective effects. The neuroprotective mechanisms may be associated with modulation of intracellular signaling cascades such as the phosphatidyl inositol 3-kinase (PI3K) pathway. Using differentiated SH-SY5Y cells, this study investigated whether BB modulation of intracellular signaling pathways, such as the protein kinase C

Chun Shi; Fengming Wu; Jie Xu; Juntao Zou

2011-01-01

9

Possible mechanism for preterm labor associated with bacterial infection. I. Stimulation of phosphoinositide metabolism by endotoxin in endometrial fibroblasts  

SciTech Connect

Growing evidence suggests an association between intra-amniotic infection and premature initiation of parturition. We recently demonstrated that some factor(s) including endotoxin produced by the organism stimulates endogenous phospholipase A2 resulting in liberation of arachidonic acid and prostaglandin formation. The studies presented in this report were designated to evaluate the mechanism for endotoxin to stimulate phospholipase A2 using human endometrial fibroblasts. Exposure of the fibroblasts to endotoxin from Escherichia coli in the presence of ({sup 32}P) phosphate increased {sup 32}P-labeling of phosphatidic acid (PA) and phosphatidyl-inositol (PI) in a dose-dependent and a time-dependent manners. The PA labeling occurred without a measurable lag time. These findings demonstrate that the endotoxin stimulates phosphoinositide metabolism in human endometrial fibroblasts by a receptor-mediated mechanism. Membrane phosphoinositide turnover stimulated by endotoxin results in cytosolic Ca{sup 2+} increment, liberation of arachidonic acid, which may be involved in the initiation of parturition.

Khan, A.A.; Imai, A.; Tamaya, T. (Gifu Univ. School of Medicine (Japan))

1990-07-01

10

Phosphoinositides as Regulators of Protein-Chromatin Interactions  

NSDL National Science Digital Library

The molecular function of phospholipids in the nucleus has been only partially elucidated. The upsurge of epigenetic research has contributed to increased interest in nuclear phospholipids, such as phosphoinositides, and their involvement in gene transcription. However, the mechanisms by which phosphoinositides regulate transcription is still unknown at the molecular level. Certain phosphoinositide species can regulate protein-chromatin and protein–nucleic acid interactions, and specific nuclear target proteins link nuclear signaling lipids to gene expression. We propose that a phosphoinositide-mediated detachment of proteins from chromatin is a general biological mechanism that partly underlies the signaling effects of nuclear phosphoinositides.

Keijo Viiri (School of Medicine and Tampere University Hospital;University of Tampere REV); Markku Maki (School of Medicine and Tampere University Hospital;University of Tampere REV); Olli Lohi (School of Medicine and Tampere University Hospital;University of Tampere REV)

2012-05-01

11

Phosphoinositide phosphatases and disease  

PubMed Central

The field of inositol signaling has expanded greatly in recent years. Given the many reviews on phosphoinositide kinases, we have chosen to restrict our discussion to inositol lipid hydrolysis focused on the phosphatases and a brief mention of the lipase isoforms. We also discuss recent discoveries that link mutations in phosphoinositide phosphatases to disease. PMID:19001665

Majerus, Philip W.; York, John D.

2009-01-01

12

Apoptosis Suppression by Raf-1 and MEK1 Requires MEK- and Phosphatidylinositol 3-Kinase-Dependent Signals  

PubMed Central

Two Ras effector pathways leading to the activation of Raf-1 and phosphatidylinositol 3-kinase (PI3K) have been implicated in the survival signaling by the interleukin 3 (IL-3) receptor. Analysis of apoptosis suppression by Raf-1 demonstrated the requirement for mitochondrial translocation of the kinase in this process. This could be achieved either by overexpression of the antiapoptotic protein Bcl-2 or by targeting Raf-1 to the mitochondria via fusion to the mitochondrial protein Mas p70. Mitochondrially active Raf-1 is unable to activate extracellular signal-related kinase 1 (ERK1) and ERK2 but suppresses cell death by inactivating the proapoptotic Bcl-2 family member BAD. However, genetic and biochemical data also have suggested a role for the Raf-1 effector module MEK-ERK in apoptosis suppression. We thus tested for MEK requirement in cell survival signaling using the interleukin 3 (IL-3)-dependent cell line 32D. MEK is essential for survival and growth in the presence of IL-3. Upon growth factor withdrawal the expression of constitutively active MEK1 mutants significantly delays the onset of apoptosis, whereas the presence of a dominant negative mutant accelerates cell death. Survival signaling by MEK most likely results from the activation of ERKs since expression of a constitutively active form of ERK2 was as effective in protecting NIH 3T3 fibroblasts against doxorubicin-induced cell death as oncogenic MEK. The survival effect of activated MEK in 32D cells is achieved by both MEK- and PI3K-dependent mechanisms and results in the activation of PI3K and in the phosphorylation of AKT. MEK and PI3K dependence is also observed in 32D cells protected from apoptosis by oncogenic Raf-1. Additionally, we also could extend these findings to the IL-3-dependent pro-B-cell line BaF3, suggesting that recruitment of MEK is a common mechanism for survival signaling by activated Raf. Requirement for the PI3K effector AKT in this process is further demonstrated by the inhibitory effect of a dominant negative AKT mutant on Raf-1-induced cell survival. Moreover, a constitutively active form of AKT synergizes with Raf-1 in apoptosis suppression. In summary these data strongly suggest a Raf effector pathway for cell survival that is mediated by MEK and AKT. PMID:11259582

von Gise, Alexander; Lorenz, Petra; Wellbrock, Claudia; Hemmings, Brian; Berberich-Siebelt, Friederike; Rapp, Ulf R.; Troppmair, Jakob

2001-01-01

13

Phosphatidylinositol 3-Kinase dependent upregulation of the epidermal growth factor receptor upon Flotillin-1 depletion in breast cancer cells  

PubMed Central

Background Flotillin-1 and flotillin-2 are two homologous and ubiquitously expressed proteins that are involved in signal transduction and membrane trafficking. Recent studies have reported that flotillins promote breast cancer progression, thus making them interesting targets for breast cancer treatment. In the present study, we have investigated the underlying molecular mechanisms of flotillins in breast cancer. Methods Human adenocarcinoma MCF7 breast cancer cells were stably depleted of flotillins by means of lentivirus mediated short hairpin RNAs. Western blotting, immunofluorescence and quantitative real-time PCR were used to analyze the expression of proteins of the epidermal growth factor receptor (EGFR) family. Western blotting was used to investigate the effect of EGFR stimulation or inhibition as well as phosphatidylinositol 3-kinase (PI3K) inhibition on mitogen activated protein kinase (MAPK) signaling. Rescue experiments were performed by stable transfection of RNA intereference resistant flotillin proteins. Results We here show that stable knockdown of flotillin-1 in MCF7 cells resulted in upregulation of EGFR mRNA and protein expression and hyperactivation of MAPK signaling, whereas ErbB2 and ErbB3 expression were not affected. Treatment of the flotillin knockdown cells with an EGFR inhibitor reduced the MAPK signaling, demonstrating that the increased EGFR expression and activity is the cause of the increased signaling. Stable ectopic expression of flotillins in the knockdown cells reduced the increased EGFR expression, demonstrating a direct causal relationship between flotillin-1 expression and EGFR amount. Furthermore, the upregulation of EGFR was dependent on the PI3K signaling pathway which is constitutively active in MCF7 cells, and PI3K inhibition resulted in reduced EGFR expression. Conclusions This study demonstrates that flotillins may not be suitable as cancer therapy targets in cells that carry certain other oncogenic mutations such as PI3K activating mutations, as unexpected effects are prone to emerge upon flotillin knockdown which may even facilitate cancer cell growth and proliferation. PMID:24304721

2013-01-01

14

Adapter protein with a pleckstrin homology (PH) and an Src homology 2 (SH2) domain (APS) and SH2-B enhance insulin-receptor autophosphorylation, extracellular-signal-regulated kinase and phosphoinositide 3-kinase-dependent signalling.  

PubMed

Adapter protein with a pleckstrin homology (PH) and an Src homology 2 (SH2) domain (APS) and SH2-B are adapter proteins and substrates that interact with the activation loop of the insulin-receptor (IR) kinase. These proteins are homologous and share substantial sequence similarity. We previously showed [Ahmed, Smith and Pillay, FEBS Lett. 475, 31-34], for the first time, that insulin-stimulated phosphorylation of APS led to interaction with c-Cbl in 3T3-L1 adipocytes and in transfected Chinese-hamster ovary (CHO) cells. In the present study, we find that insulin stimulates the membrane translocation and phosphorylation of APS to a much greater extent than SH2-B, despite the structural similarity of these proteins. Expression of APS or SH2-B delays IR tyrosine and IR substrate (IRS) dephosphorylation. This enhancement of signalling is also observed downsteam of the receptor. In control cells that lack APS, following insulin stimulation, extracellular-signal-regulated kinase (ERK) and Akt kinase reach maximal activation and then decline to basal levels by 60 min. In contrast, in APS- and SH2-B-expressing cells, ERK and Akt kinase activation remains at peak levels at 60 min. These effects may occur because these proteins either stabilize the active conformation or prevent dephosphorylation of the IR. We therefore conclude that, despite the ability to couple to c-Cbl, APS functions as a positive regulator of IR signalling and, although SH2-B is a poor substrate for the IR, its association with the IR allows it to regulate pathways downstream of the receptor independently of its phosphorylation. PMID:12521378

Ahmed, Zamal; Pillay, Tahir S

2003-04-15

15

Early activation of mTORC1 signalling in response to mechanical overload is independent of phosphoinositide 3-kinase/Akt signalling.  

PubMed

The mammalian target of rapamycin complex 1 (mTORC1) functions as a central integrator of a wide range of signals that modulate protein metabolism and cell growth. However, the contributions of individual pathways regulating mTORC1 activity in skeletal muscle are poorly defined. The purpose of this study was to determine the regulatory mechanisms that contribute to mTORC1 activation during mechanical overload-induced skeletal muscle hypertrophy. Consistent with previous studies, mechanical overload induced progressive hypertrophy of the plantaris muscle which was associated with significant increases in total RNA content and protein metabolism. mTORC1 was activated after a single day of overload as indicated by a significant increase in S6K1 phosphorylation at T389 and T421/S424. In contrast, Akt activity, as assessed by Akt phosphorylation status (T308 and S473), phosphorylation of direct downstream targets (glycogen synthase kinase 3 ?, proline-rich Akt substrate 40 kDa and tuberous sclerosis 2 (TSC2)) and a kinase assay, was not significantly increased until 2–3 days of overload. Inhibition of phosphoinositide 3-kinase (PI3K) activity by wortmannin was sufficient to block insulin-dependent signalling but did not prevent the early activation of mTORC1 in response to overload. We identified that the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK)-dependent pathway was activated at day 1 after overload. In addition, a target of MEK/ERK signalling, phosphorylation of TSC2 at S664, was also increased at this early time point. These observations demonstrate that in vivo, mTORC1 activation at the early phase of mechanical overload in skeletal muscle occurs independently of PI3K/Akt signalling and provide evidence that the MEK/ERK pathway may contribute to mTORC1 activation through phosphorylation of TSC2. PMID:21300751

Miyazaki, Mitsunori; McCarthy, John J; Fedele, Mark J; Esser, Karyn A

2011-04-01

16

Phosphoinositide signalling in cancer: beyond PI3K and PTEN.  

PubMed

There are numerous studies that suggest multiple links between the cellular phosphoinositide system and cancer. As key roles in cancer have been established for PI3K and PTEN - enzymes that regulate the levels of phosphatidylinositol-3,4,5-trisphosphate - compounds targeting this pathway are entering the clinic at a rapid pace. Several other phosphoinositide-modifying enzymes, including phosphoinositide kinases, phosphatases and phospholipase C enzymes, have been implicated in the generation and progression of tumours. Studies of these enzymes are providing new insights into the mechanisms and the extent of their involvement in cancer, highlighting new potential targets for therapeutic intervention. PMID:20414202

Bunney, Tom D; Katan, Matilda

2010-05-01

17

Phosphoinositides alter lipid bilayer properties  

PubMed Central

Phosphatidylinositol-4,5-bisphosphate (PIP2), which constitutes ?1% of the plasma membrane phospholipid, plays a key role in membrane-delimited signaling. PIP2 regulates structurally and functionally diverse membrane proteins, including voltage- and ligand-gated ion channels, inwardly rectifying ion channels, transporters, and receptors. In some cases, the regulation is known to involve specific lipid–protein interactions, but the mechanisms by which PIP2 regulates many of its various targets remain to be fully elucidated. Because many PIP2 targets are membrane-spanning proteins, we explored whether the phosphoinositides might alter bilayer physical properties such as curvature and elasticity, which would alter the equilibrium between membrane protein conformational states—and thereby protein function. Taking advantage of the gramicidin A (gA) channels’ sensitivity to changes in lipid bilayer properties, we used gA-based fluorescence quenching and single-channel assays to examine the effects of long-chain PIP2s (brain PIP2, which is predominantly 1-stearyl-2-arachidonyl-PIP2, and dioleoyl-PIP2) on bilayer properties. When premixed with dioleoyl-phosphocholine at 2 mol %, both long-chain PIP2s produced similar changes in gA channel function (bilayer properties); when applied through the aqueous solution, however, brain PIP2 was a more potent modifier than dioleoyl-PIP2. Given the widespread use of short-chain dioctanoyl-phosphoinositides, we also examined the effects of diC8-phosphoinositol (PI), PI(4,5)P2, PI(3,5)P2, PI(3,4)P2, and PI(3,4,5)P3. The diC8 phosphoinositides, except for PI(3,5)P2, altered bilayer properties with potencies that decreased with increasing head group charge. Nonphosphoinositide diC8 phospholipids generally were more potent bilayer modifiers than the polyphosphoinositides. These results show that physiological increases or decreases in plasma membrane PIP2 levels, as a result of activation of PI kinases or phosphatases, are likely to alter lipid bilayer properties, in addition to any other effects they may have. The results further show that exogenous PIP2, as well as structural analogues that differ in acyl chain length or phosphorylation state, alters lipid bilayer properties at the concentrations used in many cell physiological experiments. PMID:23712549

Hobart, E. Ashley; Koeppe, Roger E.; Andersen, Olaf S.

2013-01-01

18

Phosphoinositides and engulfment.  

PubMed

Cellular engulfment of particles, cells or solutes displaces large domains of plasma membrane into intracellular membranous vacuoles. This transfer of membrane is accompanied by major transitions of the phosphoinositide (PI) species that comprise the cytoplasmic face of membrane bilayers. Mapping of membrane PIs during engulfment reveals distinct patterns of protein and PI distributions associated with each stage of engulfment, which correspond with activities that regulate the actin cytoskeleton, membrane movements and vesicle secretion. Experimental manipulation of PI chemistry during engulfment indicates that PIs integrate organelle identity and orient signal transduction cascades within confined subdomains of membrane. These pathways are exploited by microbial pathogens to direct or redirect the engulfment process. PMID:25073505

Swanson, Joel A

2014-10-01

19

Phosphoinositides and vesicular membrane traffic  

PubMed Central

Phosphoinositide lipids were initially discovered as precursors for specific second messengers involved in signal transduction, but have now taken the center stage in controlling many essential processes at virtually every cellular membrane. In particular, phosphoinositides play a critical role in regulating membrane dynamics and vesicular transport. The unique distribution of certain phosphoinositides at specific intracellular membranes makes these molecules uniquely suited to direct organelle-specific trafficking reactions. In this regulatory role, phosphoinositides cooperate specifically with small GTPases from the Arf and Rab families. This review will summarize recent progress in the study of phosphoinositides in membrane trafficking and organellar organization and highlight the particular relevance of these signaling pathways in disease. PMID:22281700

Mayinger, Peter

2012-01-01

20

The role of phosphoinositides in synapse function.  

PubMed

Since the discovery of phosphatidylinositol-3-kinase, scientific interest in the biological functions of phosphoinositides has greatly increased. Currently, seven phosphoinositides have been identified. These phosphoinositides are specifically localized to organelle membranes, their site of action. Phosphoinositides can regulate neuronal function by specifically recruiting downstream proteins that have phosphoinositide-binding domains. To date, it is well accepted that phosphoinositides play important roles in a broad spectrum of neuronal functions from regulating neural development to modulating synapse function. This review will provide an overview of the function and distribution of phosphoinositides at synapses. PMID:24935718

Ueda, Yoshibumi

2014-12-01

21

Endosomal Phosphoinositides and Human Diseases  

PubMed Central

Phosphoinositides (PIs) are lipid second messengers implicated in signal transduction and membrane trafficking. Seven distinct PIs can be synthesized by phosphorylation of the inositol ring of phosphatidylinositol (PtdIns), and their metabolism is accurately regulated by PI kinases and phosphatases. Two of the PIs, PtdIns3P and PtdIns(3,5)P2, are present on intracellular endosomal compartments, and several studies suggest that they have a role in membrane remodeling and trafficking. We refer to them as ‘endosomal PIs’. An increasing number of human genetic diseases including myopathy and neuropathies are associated to mutations in enzymes regulating the turnover of these endosomal PIs. The PtdIns3P and PtdIns(3,5)P2 3-phosphatase myotubularin gene is mutated in X-linked centronuclear myopathy, whereas its homologs MTMR2 and MTMR13 and the PtdIns(3,5)P2 5-phosphatase SAC3/FIG4 are implicated in Charcot–Marie–Tooth peripheral neuropathies. Mutations in the gene encoding the PtdIns3P5-kinase PIP5K3/PIKfyve have been found in patients affected with François–Neetens fleck corneal dystrophy. This review presents the roles of the endosomal PIs and their regulators and proposes defects of membrane remodeling as a common pathological mechanism for the corresponding diseases. PMID:18429927

Nicot, Anne-Sophie; Laporte, Jocelyn

2008-01-01

22

Endosomal phosphoinositides and human diseases.  

PubMed

Phosphoinositides (PIs) are lipid second messengers implicated in signal transduction and membrane trafficking. Seven distinct PIs can be synthesized by phosphorylation of the inositol ring of phosphatidylinositol (PtdIns), and their metabolism is accurately regulated by PI kinases and phosphatases. Two of the PIs, PtdIns3P and PtdIns(3,5)P(2), are present on intracellular endosomal compartments, and several studies suggest that they have a role in membrane remodeling and trafficking. We refer to them as 'endosomal PIs'. An increasing number of human genetic diseases including myopathy and neuropathies are associated to mutations in enzymes regulating the turnover of these endosomal PIs. The PtdIns3P and PtdIns(3,5)P(2) 3-phosphatase myotubularin gene is mutated in X-linked centronuclear myopathy, whereas its homologs MTMR2 and MTMR13 and the PtdIns(3,5)P(2) 5-phosphatase SAC3/FIG4 are implicated in Charcot-Marie-Tooth peripheral neuropathies. Mutations in the gene encoding the PtdIns3P 5-kinase PIP5K3/PIKfyve have been found in patients affected with François-Neetens fleck corneal dystrophy. This review presents the roles of the endosomal PIs and their regulators and proposes defects of membrane remodeling as a common pathological mechanism for the corresponding diseases. PMID:18429927

Nicot, Anne-Sophie; Laporte, Jocelyn

2008-08-01

23

Arf6-independent GPI-anchored Protein-enriched Early Endosomal Compartments Fuse with Sorting Endosomes via a Rab5/Phosphatidylinositol-3?-Kinase–dependent Machinery  

PubMed Central

In the process of internalization of molecules from the extracellular milieu, a cell uses multiple endocytic pathways, consequently generating different endocytic vesicles. These primary endocytic vesicles are targeted to specific destinations inside the cell. Here, we show that GPI-anchored proteins are internalized by an Arf6-independent mechanism into GPI-anchored protein-enriched early endosomal compartments (GEECs). Internalized GPI-anchored proteins and the fluid phase are first visualized in GEECs that are acidic, primary endocytic structures, negative for early endosomal markers, Rab4, Rab5, and early endosome antigen (EEA)1. They subsequently acquire Rab5 and EEA1 before homotypic fusion with other GEECs, and heterotypic fusion with endosomes containing cargo from the clathrin-dependent endocytic pathway. Although, the formation of GEECs is unaffected by inhibition of Rab5 GTPase and phosphatidylinositol-3?-kinase (PI3K) activity, their fusion with sorting endosomes is dependent on both activities. Overexpression of Rab5 reverts PI3K inhibition of fusion, providing evidence that Rab5 effectors play important roles in heterotypic fusion between the dynamin-independent GEECs and clathrin- and dynamin-dependent sorting endosomes. PMID:16760436

Kalia, Manjula; Kumari, Sudha; Chadda, Rahul; Hill, Michelle M.; Parton, Robert G.

2006-01-01

24

The anti-apoptotic effect of IGF-1 on tissue resident stem cells is mediated via PI3-kinase dependent secreted frizzled related protein 2 (Sfrp2) release  

SciTech Connect

Previous studies suggest that IGF-1 may be used as an adjuvant to stem cell transfer in order to improve cell engraftment in ischemic tissue. In the current study, we investigated the effect of IGF-1 on serum deprivation and hypoxia induced stem cell apoptosis and the possible mechanisms involved. Exposure of adipose tissue derived stem cells (ASCs) to serum deprivation and hypoxia resulted in significant apoptosis in ASC which is partially prevented by IGF-1. IGF-1's anti-apoptotic effect was abolished in ASCs transfected with Sfrp2 siRNA but not by the control siRNA. Using Western blot analysis, we demonstrated that serum deprivation and hypoxia reduced the expression of nuclear {beta}-catenin, which is reversed by IGF-1. IGF-1's effect on {beta}-catenin expression was abolished by the presence of PI3-kinase inhibitor LY294002 or in ASCs transfected with Sfrp2 siRNA. These results suggest that IGF-1, through the release of the Sfrp2, contributes to cell survival by stabilizing {beta}-catenin.

Gehmert, Sebastian; Sadat, Sanga; Song Yaohua; Yan Yasheng [Department of Molecular Pathology, University of Texas M.D. Anderson Cancer Center, SCRB2, Box 951, 7435 Fannin Street, Houston, TX 77030 (United States); Alt, Eckhard [Department of Molecular Pathology, University of Texas M.D. Anderson Cancer Center, SCRB2, Box 951, 7435 Fannin Street, Houston, TX 77030 (United States)], E-mail: ealt@mdanderson.org

2008-07-11

25

Prostaglandin E2 induces interleukin-6 expression in human chondrocytes via cAMP/protein kinase A- and phosphatidylinositol 3-kinase-dependent NF-?B activation  

PubMed Central

Elevated levels of prostaglandin (PG)E2 and interleukin (IL)-6 have been reported in the cartilage and synovial fluid from patients with arthritic disorders. PGE2 regulates IL-6 production in numerous different cells including macrophages and synovial fibroblasts. Although PGE2 stimulates IL-6 expression in human chondrocytes, the underlying signaling pathway of this process has yet to be delineated. Here, we investigate the mechanism of IL-6 induction in human T/C-28a2 chondrocytes treated with exogenously added PGE2. PGE2 induces IL-6 mRNA and protein expression via a cAMP-dependent pathway, reaching maximal levels after 60 min of stimulation before declining to baseline levels at 6 h. Forskolin, an adenylyl cyclase activator, also stimulates IL-6 expression in human chondrocytes in a dose- and time-dependent fashion. Inhibition of downstream effectors of cAMP activity such as protein kinase A (PKA) or phosphatidylinositol 3 kinase (PI3K) blocks PGE2- and forskolin-induced IL-6 upregulation. Simultaneous inhibition of PKA and PI3K reduces IL-6 expression in stimulated chondrocytes well below the basal levels of untreated cells. Gel shift, supershift, and chromatin immunoprecipitation assays reveal the activation and binding of the nuclear factor (NF)-?B p65 subunit to the IL-6 promoter, which is markedly suppressed by selective PI3K or PKA pharmacological inhibitors. p65 knockdown completely abrogates IL-6 mRNA synthesis in PGE2- and forskolin-primed chondrocytes. Cumulatively, our data show that PGE2 and forskolin induce IL-6 expression in human chondrocytes via cAMP/PKA and PI3K-dependent pathways, which in turn regulate the activation and binding of p65 to the IL-6 promoter. PMID:20457835

Wang, Pu; Zhu, Fei

2010-01-01

26

Plant phosphoinositide-specific phospholipase C  

PubMed Central

Phosphoinositide-specific phospholipase C (PI-PLC) belongs to an important class of enzymes involved in signaling related to lipids. They hydrolyze a membrane-associated phospholipid, phosphatidylinositol-4,5-bisphosphate, to produce inositol-1,4,5-trisphosphate and diacylglycerol. The role of PI-PLC and the mechanism behind its functioning is well studied in animal system; however, mechanism of plant PI-PLC functioning remains largely obscure. Here, we attempted to summarize the understanding regarding plant PI-PLC mechanism of regulation, localization, and domain association. Using sedimentation based phospholipid binding assay and surface plasmon resonance spectroscopy, it was demonstrated that C2 domain of plant PI-PLC alone is capable of targeting membranes. Moreover, change in surface hydrophobicity upon calcium stimulus is the key element in targeting plant PI-PLC from soluble fractions to membranes. This property of altering surface hydrophobicity plays a pivot role in regulation of PI-PLC activity. PMID:22902702

Rupwate, Sunny D.; Rajasekharan, Ram

2012-01-01

27

Ion Induced Changes in Phosphoinositide Monolayers at Phisiological Concentrations  

NASA Astrophysics Data System (ADS)

Phosphoinositides (PIPs) play a crucial role in many cellular process that occur at the plasma membrane such as calcium release, exocytosis or endocytosis. In order to specifically regulate these functions PIPs must segregate in pools at the plasma membrane. A possible mechanism that could induce and regulate such organization of phosphoinositides is their interaction with bivalent cations. Understanding the physicochemical mechanism that can regulate membrane structure is a crucial step in the development of adaptive biomimetic membrane systems. Using Langmuir monolayers, we investigated the effect of calcium and magnesium on the surface pressure-area/lipid isotherm of monolayer of phosphatidylinositol (PI), phosphatidylinositol bisphosphate (PIP2), dioleoylphosphatidylglycerol (DOPG) and palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). It is found that the decrease of area per lipid, i.e. the increase in aggregation, is mostly dependent on the lipid's head group charge but ion specific. In addition, we discuss changes in free energy and compressibility of these monolayer-ion systems.

Kazadi Badiambile, Adolphe; Forstner, Martin B.

2013-03-01

28

The Phox homology (PX) domain, a new player in phosphoinositide signalling.  

PubMed Central

Phosphoinositides are key regulators of diverse cellular processes. The pleckstrin homology (PH) domain mediates the action of PtdIns(3,4)P(2), PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3), while the FYVE domain relays the pulse of PtdIns3P. The recent establishment that the Phox homology (PX) domain interacts with PtdIns3P and other phosphoinositides suggests another mechanism by which phosphoinositides can regulate/integrate multiple cellular events via a spectrum of PX domain-containing proteins. Together with the recent discovery that the epsin N-terminal homologue (ENTH) domain interacts with PtdIns(4,5)P(2), it is becoming clear that phosphoinositides regulate diverse cellular events through interactions with several distinct structural motifs present in many different proteins. PMID:11736640

Xu, Y; Seet, L F; Hanson, B; Hong, W

2001-01-01

29

Molecular basis for substrate recognition by MTMR2, a myotubularin family phosphoinositide phosphatase  

PubMed Central

Myotubularins, a large family of catalytically active and inactive proteins, belong to a unique subgroup of protein tyrosine phosphatases that use inositol phospholipids, rather than phosphoproteins, as physiological substrates. Here, by integrating crystallographic and deuterium-exchange mass spectrometry studies of human myotubularin-related protein-2 (MTMR2) in complex with phosphoinositides, we define the molecular basis for this unique substrate specificity. Phosphoinositide substrates bind in a pocket located on a positively charged face of the protein, suggesting an electrostatic mechanism for membrane targeting. A flexible, hydrophobic helix makes extensive interactions with the diacylglycerol moieties of substrates, explaining the specificity for membrane-bound phosphoinositides. An extensive H-bonding network and charge–charge interactions within the active site pocket determine phosphoinositide headgroup specificity. The conservation of these specificity determinants within the active, but not the inactive, myotubularins provides insight into the functional differences between the active and inactive members. PMID:16410353

Begley, Michael J.; Taylor, Gregory S.; Brock, Melissa A.; Ghosh, Partho; Woods, Virgil L.; Dixon, Jack E.

2006-01-01

30

Signal transduction abnormalities in suicide: focus on phosphoinositide signaling system.  

PubMed

Suicide is a major public health concern and each year about one million people die by suicide worldwide. Recent studies suggest that suicide may be associated with specific neurobiological abnormalities. Earlier studies of neurobiology of suicide focused on abnormalities of the serotonergic mechanism. These studies suggested that some serotonin receptor subtypes may be abnormal in the postmortem brain of suicide victims. Since these receptors are linked to signal transduction pathways, abnormalities of signaling mechanisms have been recently studied in the postmortem brain of suicide victims. Of particular interest is the 5-hydroxytryptamine2A receptor-linked phosphoinositide signaling system. Several studies have focused on the abnormalities on the component of this signaling system and these studies suggest the abnormalities of G proteins, the effectors phospholipase C and the second or the third messenger systems, such as protein kinase A. Further studies revealed abnormalities in the downstream transcription factors such as the cyclic AMP response element binding protein and some of the targeted genes of these transcription factors. The most important gene in this aspect which has been studied in the suicide is the brain-derived neurotrophic factor. Here we critically review the studies focusing on these components of the phosphoinositide signaling system in the postmortem brain of both adult and teenage suicide victims. These studies provide a better understanding of the signal transduction abnormalities in suicide focusing on the phosphoinositide signaling pathway. These studies may lead to new therapeutic agents targeting specific sites in this signaling cascade. PMID:24040801

Pandey, Ghanshyam N

2013-11-01

31

Mutations in Phosphoinositide Metabolizing Enzymes and Human Disease  

NSDL National Science Digital Library

Phosphoinositides are implicated in the regulation of a wide variety of cellular functions. Their importance in cellular and organismal physiology is underscored by the growing number of human diseases linked to perturbation of kinases and phosphatases that catalyze interconversion from one phosphoinositide to another. Many such enzymes are attractive targets for therapeutic interventions. Here, we review diseases linked to inheritable or somatic mutations of these enzymes. Phosphatidylinositol (PtdIns), a membrane phospholipid, can be reversibly phosphorylated at the 3, 4, and 5 positions of the inositol ring to generate seven phosphoinositides [PI3P, PI4P, PI5P, PI(3,4)P2, PI(4,5)P2, PI(3,5)P2, and PI(3,4,5)P3] (FIGURE 1A). The importance of this metabolism in cell regulation was first established in the context of studies on stimulus-secretion coupling. It was found that many stimuli that trigger secretion also trigger enhanced turnover of PtdIns and phosphoinositides (42). Subsequently, it became clear that phospholipase C-dependent hydrolysis of PI(4,5)P2 to generate the second messenger molecules diacyl glycerol and Ins(1,4,5)P3 (IP3) is a mechanism through which many cell surface receptors, including many receptors that stimulate secretion, transduce their signals (10). Diacyl glycerol binds and regulates protein kinase C and a variety of other effectors, whereas IP3 triggers calcium release from the endoplasmic reticulum (10, 42). In another signal transduction pathway, PI(4,5)P2 is cleaved by phospholipase A2 to generate arachidonic acid, a precursor of many signaling molecules.

Heather J. McCrea (Yale University); Pietro De Camilli (Yale University School of Medicine)

2009-02-01

32

Proline 326 in the C terminus of murine CX3CR1 prevents G-protein and phosphatidylinositol 3-kinase-dependent stimulation of Akt and extracellular signal-regulated kinase in Chinese hamster ovary cells.  

PubMed

Naturally occurring single nucleotide polymorphisms have been identified in human CX3CR1, the chemokine receptor for fractalkine (FKN/CX3CL1). Individuals carrying the I249/M280 variant of CX3CR1 have a lower risk of cardiovascular disease compared with those homozygous for the common variant (V249/T280). The precise molecular basis for this phenotype is unclear, although differences in FKN binding, adhesive properties, and signaling efficiency between the CX3CR1 variants have been reported. FKN binding to CX3CR1 leads to an increase in intracellular calcium, actin rearrangement, and activation of the mitogen-activated protein kinase and phosphoinositide 3-kinase (PI3K) pathways. Regulation of these signaling pathways underlies the known roles for FKN in cell survival, proliferation, and migration. In the present study, we demonstrate that FKN stimulates phosphorylation of protein kinase B (Akt/PKB) in Chinese hamster ovary cells individually expressing the naturally occurring variants of human CX3CR1-, as well as rat CX3CR1-, but not in murine CX3CR1-expressing cells. Substitution of Pro326 in the C terminus of murine CX3CR1 with Ser (residue found in the analogous position of human CX3CR1) produced a mutant receptor that mimicked the human receptor in its ability to stimulate the phosphorylation of both Akt and extracellular signal-regulated kinase in a time-, PI3K-, and pertussis toxin-sensitive G-protein-dependent manner. These results identify a critical structural determinant of CX3CR1 important for activation of downstream signaling pathways. PMID:16166268

Davis, Christopher N; Harrison, Jeffrey K

2006-01-01

33

Optogenetic control of phosphoinositide metabolism.  

PubMed

Phosphoinositides (PIs) are lipid components of cell membranes that regulate a wide variety of cellular functions. Here we exploited the blue light-induced dimerization between two plant proteins, cryptochrome 2 (CRY2) and the transcription factor CIBN, to control plasma membrane PI levels rapidly, locally, and reversibly. The inositol 5-phosphatase domain of OCRL (5-ptase(OCRL)), which acts on PI(4,5)P(2) and PI(3,4,5)P(3), was fused to the photolyase homology region domain of CRY2, and the CRY2-binding domain, CIBN, was fused to plasma membrane-targeting motifs. Blue-light illumination (458-488 nm) of mammalian cells expressing these constructs resulted in nearly instantaneous recruitment of 5-ptase(OCRL) to the plasma membrane, where it caused rapid (within seconds) and reversible (within minutes) dephosphorylation of its targets as revealed by diverse cellular assays: dissociation of PI(4,5)P(2) and PI(3,4,5)P(3) biosensors, disappearance of endocytic clathrin-coated pits, nearly complete inhibition of KCNQ2/3 channel currents, and loss of membrane ruffling. Focal illumination resulted in local and transient 5-ptase(OCRL) recruitment and PI(4,5)P(2) dephosphorylation, causing not only local collapse and retraction of the cell edge or process but also compensatory accumulation of the PI(4,5)P(2) biosensor and membrane ruffling at the opposite side of the cells. Using the same approach for the recruitment of PI3K, local PI(3,4,5)P(3) synthesis and membrane ruffling could be induced, with corresponding loss of ruffling distally to the illuminated region. This technique provides a powerful tool for dissecting with high spatial-temporal kinetics the cellular functions of various PIs and reversibly controlling the functions of downstream effectors of these signaling lipids. PMID:22847441

Idevall-Hagren, Olof; Dickson, Eamonn J; Hille, Bertil; Toomre, Derek K; De Camilli, Pietro

2012-08-28

34

Decoding the role of phosphoinositides in phototropin signaling involved in chloroplast movements  

PubMed Central

In angiosperms, light-dependent chloroplast movements are exclusively mediated by UVA/blue light receptors - phototropins. The two photoreceptors of Arabidopsis thaliana, phot1 and phot2, have overlapping roles in the control of these movements. Experiments performed in different plant species point to the participation of phosphoinositides in blue light-controlled chloroplast relocations. Here, we report a summary of recent findings presenting the involvement of phosphatidylinositol 4,5-bisphosphate as well as phosphatidylinositol 3- and 4-phosphates in weak blue light-mediated (accumulation) and strong blue light-mediated (avoidance) responses of chloroplasts. The blue light-activated alterations in phosphoinositide concentration are partly responsible for cytosolic Ca2+ changes. Ca2+ influx from apoplast does not seem to be involved in the mechanism of movement responses. In summary, interplay between phosphoinositides and intracellular Ca2+ regulates chloroplast redistribution in response to blue light in higher plants. PMID:23733070

Aggarwal, Chhavi; Labuz, Justyna; Gabrys, Halina

2013-01-01

35

CDP-diacylglycerol synthetase-controlled phosphoinositide availability limits VEGFA signaling and vascular morphogenesis  

PubMed Central

Understanding the mechanisms that regulate angiogenesis and translating these into effective therapies are of enormous scientific and clinical interests. In this report, we demonstrate the central role of CDP-diacylglycerol synthetase (CDS) in the regulation of VEGFA signaling and angiogenesis. CDS activity maintains phosphoinositide 4,5 bisphosphate (PIP2) availability through resynthesis of phosphoinositides, whereas VEGFA, mainly through phospholipase C?1, consumes PIP2 for signal transduction. Loss of CDS2, 1 of 2 vertebrate CDS enzymes, results in vascular-specific defects in zebrafish in vivo and failure of VEGFA-induced angiogenesis in endothelial cells in vitro. Absence of CDS2 also results in reduced arterial differentiation and reduced angiogenic signaling. CDS2 deficit-caused phenotypes can be successfully rescued by artificial elevation of PIP2 levels, and excess PIP2 or increased CDS2 activity can promote excess angiogenesis. These results suggest that availability of CDS-controlled resynthesis of phosphoinositides is essential for angiogenesis. PMID:22649102

Pan, Weijun; Pham, Van N.; Stratman, Amber N.; Castranova, Daniel; Kamei, Makoto; Kidd, Kameha R.; Lo, Brigid D.; Shaw, Kenna M.; Torres-Vazquez, Jesus; Mikelis, Constantinos M.; Gutkind, J. Silvio; Davis, George E.

2012-01-01

36

Serotonin-stimulated phosphoinositide turnover: mediation by the S2 binding site in rat cerebral cortex but not in subcortical regions  

SciTech Connect

In rat cerebral cortex, serotonin (5-HT) stimulates phosphoinositide turnover with an EC50 of 1 microM in the presence of pargyline. The EC50 is 16-fold higher in the absence of pargyline. Selective S2 antagonists inhibit 5-HT-stimulated phosphoinositide turnover. Schild analysis of the blockade by ketanserin of the 5-HT effect gives an estimated Kd of ketanserin for the phosphoinositide-linked receptor of 11.7 nM, which agrees with the Kd (3.5 nM) of (/sup 3/H)ketanserin for the S2 site. Furthermore, MK-212, 5-HT and 5-fluorotryptamine stimulate phosphoinositide turnover with potencies that resemble their potencies at the S2 but not the S1 binding site. Of 11 agonists tested, the tryptamine derivatives tend to be more efficacious than the piperazine derivatives. The selective S1 agonist 8-hydroxy-2-(di-N-propylamino)tetralin is inactive at stimulating phosphoinositide turnover. No significant relationship exists between the regional distributions of 5-HT-stimulated phosphoinositide turnover and S2 binding sites. Furthermore, the S2 antagonist ketanserin is less potent and less efficacious in hippocampus and limbic forebrain than in cerebral cortex. These data suggest that 5-HT-stimulated phosphoinositide turnover is linked to the S2 binding site in rat cerebral cortex. However, 5-HT increases phosphoinositide turnover in subcortical regions by mechanisms other than stimulation of the S2 receptor.

Conn, P.J.; Sanders-Bush, E.

1985-07-01

37

Myotubularin phosphoinositide phosphatases in human diseases.  

PubMed

The level and turnover of phosphoinositides (PIs) are tightly controlled by a large set of PI-specific enzymes (PI kinases and phosphatases). Mammalian PI phosphatases are conserved through evolution and among this large family the dual-specificity phosphatase (PTP/DSP) are metal-independent enzymes displaying the amino acid signature Cys-X5-Arg-Thr/Ser (CX5RT/S) in their active site. Such catalytic site characterizes the myotubularin 3-phosphatases that dephosphorylate PtdIns3P and PtdIns(3,5)P? and produce PtdIns5P. Substrates of myotubularins have been implicated in endocytosis and membrane trafficking while PtdIns5P may have a role in signal transduction. As a paradox, 6 of the 14 members of the myotubularin family lack enzymatic activity and are considered as dead phosphatases. Several myotubularins have been genetically linked to human diseases: MTM1 is mutated in the congenital myopathy X-linked centronuclear or myotubular myopathy (XLCNM) and MTMR14 (JUMPY) has been linked to an autosomal form of such disease, while MTMR2 and MTMR13 are mutated in Charcot-Marie-Tooth (CMT) neuropathies. Furthermore, recent evidences from genetic association studies revealed that several other myotubularins could be associated to chronic disorders such as cancer and obesity, highlighting their importance for human health. Here, we discuss cellular and physiological roles of myotubularins and their implication in human diseases, and we present potential pathological mechanisms affecting specific tissues in myotubularin-associated diseases. PMID:23086420

Amoasii, Leonela; Hnia, Karim; Laporte, Jocelyn

2012-01-01

38

FirstRevise Phosphoinositides and Rho Proteins Spatially Regulate Actin  

E-print Network

FirstRevise Phosphoinositides and Rho Proteins Spatially Regulate Actin Polymerization to Initiate of motile cells involve the actin cytoskeleton, and regulatory mod- ules, including the phosphoinositides components (PIP1, PIP2, PIP3; PTEN, PI3K, PI5K; Cdc42, Rac, Rho; Arp2/3, and actin), their interconversions

Keshet, Leah

39

FERM Domain Phosphoinositide Binding Targets Merlin to the Membrane and Is Essential for Its Growth-Suppressive Function ?  

PubMed Central

The neurofibromatosis type 2 tumor suppressor protein, merlin, is related to the ERM (ezrin, radixin, and moesin) family of plasma membrane-actin cytoskeleton linkers. For ezrin, phosphatidylinositol 4,5-bisphosphate (PIP2) binding to the amino-terminal FERM domain is required for its conformational activation, proper subcellular localization, and function, but less is known about the role of phosphoinositide binding for merlin. Current evidence indicates that association with the membrane is important for merlin to function as a growth regulator; however, the mechanisms by which merlin localizes to the membrane are less clear. Here, we report that merlin binds phosphoinositides, including PIP2, via a conserved binding motif in its FERM domain. Abolition of FERM domain-mediated phosphoinositide binding of merlin displaces merlin from the membrane and releases it into the cytosol without altering the folding of merlin. Importantly, a merlin protein whose FERM domain cannot bind phosphoinositide is defective in growth suppression. Retargeting the mutant merlin into the membrane using a dual-acylated amino-terminal decapeptide from Fyn is sufficient to restore the growth-suppressive properties to the mutant merlin. Thus, FERM domain-mediated phosphoinositide binding and membrane association are critical for the growth-regulatory function of merlin. PMID:21402777

Mani, Timmy; Hennigan, Robert F.; Foster, Lauren A.; Conrady, Deborah G.; Herr, Andrew B.; Ip, Wallace

2011-01-01

40

New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway  

PubMed Central

The most recently discovered PTEN tumor suppressor gene has been found to be defective in a large number of human cancers. In addition, germ-line mutations in PTEN result in the dominantly inherited disease Cowden syndrome, which is characterized by multiple hamartomas and a high proclivity for developing cancer. A series of publications over the past year now suggest a mechanism by which PTEN loss of function results in tumors. PTEN appears to negatively control the phosphoinositide 3-kinase signaling pathway for regulation of cell growth and survival by dephosphorylating the 3 position of phosphoinositides. PMID:10200246

Cantley, Lewis C.; Neel, Benjamin G.

1999-01-01

41

Measurement of phosphoinositides in the zebrafish Danio rerio.  

PubMed

Phosphoinositides represent a minor fraction of the total glycerolipids in cells. Despite the fact that phosphoinositides are present in small quantities, they have crucial roles during cell signaling and in regulating numerous intracellular processes. Measuring changes in the levels of different phosphoinositides in animals is difficult, but it is essential in order to define the important functions of specific members of the phosphoinositide family. Here we detail procedures for measuring phosphoinositides in 2-days-postfertilization (2-d.p.f.) embryos in zebrafish (Danio rerio). Both in vivo radiolabeling (using [(32)P]orthophosphate) followed by thin-layer or high-performance liquid chromatography (TLC or HPLC) analysis and specific in vitro phosphorylation assays (using [(32)P]?ATP) permit the quantitative measurement of phosphoinositides. Normalization of both measurements can be achieved by the determination of total lipid phosphate in embryos. All the techniques described are relatively inexpensive and accessible to most laboratories with an interest in studying the effect of gene manipulation on phosphoinositide metabolism in zebrafish. All the procedures described herein will take up to 10 working days. PMID:23660755

Jones, David R; Ramirez, Irene Barinaga-Rementeria; Lowe, Martin; Divecha, Nullin

2013-06-01

42

The Receptor Binding Domain of Botulinum Neurotoxin Stereotype C Binds Phosphoinositides  

SciTech Connect

Botulinum neurotoxins (BoNTs) are the most toxic proteins known for humans and animals with an extremely low LD50 of {approx} 1 ng/kg. BoNTs generally require a protein and a ganglioside on the cell membrane surface for binding, which is known as a 'dual receptor' mechanism for host intoxication. Recent studies have suggested that in addition to gangliosides, other membrane lipids such as phosphoinositides may be involved in the interactions with the receptor binding domain (HCR) of BoNTs for better membrane penetration. Here, using two independent lipid-binding assays, we tested the interactions of BoNT/C-HCR with lipids in vitro. BoNT/C-HCR was found to bind negatively charged phospholipids, preferentially phosphoinositides. Additional interactions to phosphoinositides may help BoNT/C bind membrane more tightly and transduct signals for subsequent steps of intoxication. Our results provide new insights into the mechanisms of host cell membrane recognition by BoNTs.

Zhang, Yanfeng; Varnum, Susan M.

2012-03-01

43

Mutations in Phosphoinositide Metabolizing Enzymes and Human Disease  

PubMed Central

Phosphoinositides are implicated in the regulation of a wide variety of cellular functions. Their importance in cellular and organismal physiology is underscored by the growing number of human diseases linked to perturbation of kinases and phosphatases that catalyze interconversion from one phosphoinositide to another. Many such enzymes are attractive targets for therapeutic interventions. Here, we review diseases linked to inheritable or somatic mutations of these enzymes. PMID:19196647

McCrea, Heather J.; De Camilli, Pietro

2012-01-01

44

PITPs as Targets for Selectively Interfering With Phosphoinositide Signaling in Cells  

PubMed Central

Sec14-like phosphatidylinositol transfer proteins (PITPs) integrate diverse territories of intracellular lipid metabolism with stimulated phosphatidylinositol-4-phosphate production, and are discriminating portals for interrogating phosphoinositide signaling. Yet, neither Sec14-like PITPs, nor PITPs in general, have been exploited as targets for chemical inhibition for such purposes. Herein, we validate the first small molecule inhibitors (SMIs) of the yeast PITP Sec14. These SMIs are nitrophenyl(4-(2-methoxyphenyl)piperazin-1-yl)methanones (NPPMs), and are effective inhibitors in vitro and in vivo. We further establish Sec14 is the sole essential NPPM target in yeast, that NPPMs exhibit exquisite targeting specificities for Sec14 (relative to related Sec14-like PITPs), propose a mechanism for how NPPMs exert their inhibitory effects, and demonstrate NPPMs exhibit exquisite pathway selectivity in inhibiting phosphoinositide signaling in cells. These data deliver proof-of-concept that PITP-directed SMIs offer new and generally applicable avenues for intervening with phosphoinositide signaling pathways with selectivities superior to those afforded by contemporary lipid kinase-directed strategies. PMID:24292071

Nile, Aaron H.; Tripathi, Ashutosh; Yuan, Peihua; Mousley, Carl J.; Suresh, Sundari; Wallace, Iain Michael; Shah, Sweety D.; Pohlhaus, Denise Teotico; Temple, Brenda; Nislow, Corey; Giaever, Guri; Tropsha, Alexander; Davis, Ronald W.; St Onge, Robert P.; Bankaitis, Vytas A.

2013-01-01

45

Retroviruses Human Immunodeficiency Virus and Murine Leukemia Virus Are Enriched in Phosphoinositides? †  

PubMed Central

Retroviruses acquire a lipid envelope during budding from the membrane of their hosts. Therefore, the composition of this envelope can provide important information about the budding process and its location. Here, we present mass spectrometry analysis of the lipid content of human immunodeficiency virus type 1 (HIV-1) and murine leukemia virus (MLV). The results of this comprehensive survey found that the overall lipid content of these viruses mostly matched that of the plasma membrane, which was considerably different from the total lipid content of the cells. However, several lipids are enriched in comparison to the composition of the plasma membrane: (i) cholesterol, ceramide, and GM3; and (ii) phosphoinositides, phosphorylated derivatives of phosphatidylinositol. Interestingly, microvesicles, which are similar in size to viruses and are also released from the cell periphery, lack phosphoinositides, suggesting a different budding mechanism/location for these particles than for retroviruses. One phosphoinositide, phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], has been implicated in membrane binding by HIV Gag. Consistent with this observation, we found that PI(4,5)P2 was enriched in HIV-1 and that depleting this molecule in cells reduced HIV-1 budding. Analysis of mutant virions mapped the enrichment of PI(4,5)P2 to the matrix domain of HIV Gag. Overall, these results suggest that HIV-1 and other retroviruses bud from cholesterol-rich regions of the plasma membrane and exploit matrix/PI(4,5)P2 interactions for particle release from cells. PMID:18799574

Chan, Robin; Uchil, Pradeep D.; Jin, Jing; Shui, Guanghou; Ott, David E.; Mothes, Walther; Wenk, Markus R.

2008-01-01

46

Phosphoinositide metabolism and adrenergic receptors in astrocytes  

SciTech Connect

Agonist-induced phosphoinositide (PI) breakdown functions as a signal generating system. Diacylglycerol, one breakdown product of phosphotidylinositol-4,5-diphosphate hydrolysis, can stimulate protein kinase C, whereas inositol triphosphate, the other product, has been proposed to be a second messenger for Ca/sup + +/ mobilization. Using purified astrocyte cultures from neonatal rat brain, the effects of adrenergic agonists and antagonists at 10/sup -5/ M were measured on PI breakdown. Astrocytes grown in culture were prelabeled with (/sup 3/H)inositol, and basal (/sup 3/H) inositol phosphate (IP/sub 1/) accumulation was measured in the presence of Li/sup +/. Epinephrine > norepinephrine (NE) were the most active stimulants of IP/sub 1/ production. The ..cap alpha../sub 1/ adrenoreceptor blockers, phentolamine and phenoxybenzamine, added alone had no effect on IP/sub 1/ production was reduced below basal levels. Propranolol partially blocked the effects of NE. Clonidine and isoproterenol, separately added, reduced IP/sub 1/ below basal levels and when added together diminished IP/sub 1/ accumulation even further. The role of adrenergic stimulation in the production of c-AMP.

Noble, E.P.; Ritchie, T.; de Vellis, J.

1986-03-01

47

Cellular and molecular interactions of phosphoinositides and peripheral proteins.  

PubMed

Anionic lipids act as signals for the recruitment of proteins containing cationic clusters to biological membranes. A family of anionic lipids known as the phosphoinositides (PIPs) are low in abundance, yet play a critical role in recruitment of peripheral proteins to the membrane interface. PIPs are mono-, bis-, or trisphosphorylated derivatives of phosphatidylinositol (PI) yielding seven species with different structure and anionic charge. The differential spatial distribution and temporal appearance of PIPs is key to their role in communicating information to target proteins. Selective recognition of PIPs came into play with the discovery that the substrate of protein kinase C termed pleckstrin possessed the first PIP binding region termed the pleckstrin homology (PH) domain. Since the discovery of the PH domain, more than ten PIP binding domains have been identified including PH, ENTH, FYVE, PX, and C2 domains. Representative examples of each of these domains have been thoroughly characterized to understand how they coordinate PIP headgroups in membranes, translocate to specific membrane docking sites in the cell, and function to regulate the activity of their full-length proteins. In addition, a number of novel mechanisms of PIP-mediated membrane association have emerged, such as coincidence detection-specificity for two distinct lipid headgroups. Other PIP-binding domains may also harbor selectivity for a membrane physical property such as charge or membrane curvature. This review summarizes the current understanding of the cellular distribution of PIPs and their molecular interaction with peripheral proteins. PMID:24556335

Stahelin, Robert V; Scott, Jordan L; Frick, Cary T

2014-09-01

48

Phosphoinositide 3-kinase p85beta regulates invadopodium formation  

PubMed Central

ABSTRACT The acquisition of invasiveness is characteristic of tumor progression. Numerous genetic changes are associated with metastasis, but the mechanism by which a cell becomes invasive remains unclear. Expression of p85?, a regulatory subunit of phosphoinositide-3-kinase, markedly increases in advanced carcinoma, but its mode of action is unknown. We postulated that p85? might facilitate cell invasion. We show that p85? localized at cell adhesions in complex with focal adhesion kinase and enhanced stability and maturation of cell adhesions. In addition, p85? induced development at cell adhesions of an F-actin core that extended several microns into the cell z-axis resembling the skeleton of invadopodia. p85? lead to F-actin polymerization at cell adhesions by recruiting active Cdc42/Rac at these structures. In accordance with p85? function in invadopodium-like formation, p85? levels increased in metastatic melanoma and p85? depletion reduced invadopodium formation and invasion. These results show that p85? enhances invasion by inducing cell adhesion development into invadopodia-like structures explaining the metastatic potential of tumors with increased p85? levels. PMID:25217619

Cariaga-Martinez, Ariel E.; Cortes, Isabel; Garcia, Esther; Perez-Garcia, Vicente; Pajares, Maria J.; Idoate, Miguel A.; Redondo-Munoz, Javier; Anton, Ines M.; Carrera, Ana C.

2014-01-01

49

Phosphoinositide analysis by liquid chromatography-mass spectrometry.  

PubMed

The phosphoinositides are a highly dynamic group of molecules implicated in many cellular control processes; however, the analysis of many of these structures has proven very difficult and time-consuming, with limited sensitivity and/or discrimination. Recent developments in LCMS now provide the prospect of routine structural and quantitative analysis of all the known phosphoinositides (and possibly some as yet unidentified structures) at high sensitivity in any biological sample. The procedures described here give very high extraction recovery from a variety of biological matrices and enable chromatographic resolution of most phosphoinositides as their native structures. When coupled with the accurate mass and fragmentation capabilities of an MS, full structural and isomeric identification can be achieved. PMID:20645190

Pettitt, Trevor R

2010-01-01

50

Two structural components in CNGA3 support regulation of cone CNG channels by phosphoinositides  

PubMed Central

Cyclic nucleotide-gated (CNG) channels in retinal photoreceptors play a crucial role in vertebrate phototransduction. The ligand sensitivity of photoreceptor CNG channels is adjusted during adaptation and in response to paracrine signals, but the mechanisms involved in channel regulation are only partly understood. Heteromeric cone CNGA3 (A3) + CNGB3 (B3) channels are inhibited by membrane phosphoinositides (PIPn), including phosphatidylinositol 3,4,5-triphosphate (PIP3) and phosphatidylinositol 4,5-bisphosphate (PIP2), demonstrating a decrease in apparent affinity for cyclic guanosine monophosphate (cGMP). Unlike homomeric A1 or A2 channels, A3-only channels paradoxically did not show a decrease in apparent affinity for cGMP after PIPn application. However, PIPn induced an ?2.5-fold increase in cAMP efficacy for A3 channels. The PIPn-dependent change in cAMP efficacy was abolished by mutations in the C-terminal region (R643Q/R646Q) or by truncation distal to the cyclic nucleotide-binding domain (613X). In addition, A3-613X unmasked a threefold decrease in apparent cGMP affinity with PIPn application to homomeric channels, and this effect was dependent on conserved arginines within the N-terminal region of A3. Together, these results indicate that regulation of A3 subunits by phosphoinositides exhibits two separable components, which depend on structural elements within the N- and C-terminal regions, respectively. Furthermore, both N and C regulatory modules in A3 supported PIPn regulation of heteromeric A3+B3 channels. B3 subunits were not sufficient to confer PIPn sensitivity to heteromeric channels formed with PIPn-insensitive A subunits. Finally, channels formed by mixtures of PIPn-insensitive A3 subunits, having complementary mutations in N- and/or C-terminal regions, restored PIPn regulation, implying that intersubunit N–C interactions help control the phosphoinositide sensitivity of cone CNG channels. PMID:23530136

Dai, Gucan; Peng, Changhong; Liu, Chunming

2013-01-01

51

Multidrug resistance-associated protein 1 expression is under the control of the phosphoinositide 3 kinase\\/Akt signal transduction network in human acute myelogenous leukemia blasts  

Microsoft Academic Search

A high incidence of relapses following induction chemotherapy is a major hindrance to patient survival in acute myelogenous leukemia (AML). There is strong evidence that activation of the phosphoinositide 3 kinase (PI3K)\\/Akt signaling network plays a significant role in rendering AML blasts drug resistant. An important mechanism underlying drug resistance is represented by overexpression of membrane drug transporters such as

P L Tazzari; A Cappellini; F Ricci; C Evangelisti; V Papa; T Grafone; G Martinelli; R Conte; L Cocco; J A McCubrey; A M Martelli

2007-01-01

52

Soman-induced seizures impair norepinephrine-stimulated phosphoinositide turnover  

SciTech Connect

Seizure activity increases turnover of phosphoinositide bisphosphate (PIP2). Turnover of PIP2 is thought to be modulated by neurotransmitter interactions. The effect of soman-induced seizures on neurotransmitter-stimulated PIP 2 turnover was examined in rats. Thirty minutes after induction of seizure activity, rats were euthanized and slices prepared from the hippocampus or cerebral cortex were incubated with myo-(2-3H) inositol for incorporation into phospholipids. Hydrolysis of phosphoinositides was determined by measuring the accumulation of (3H) inositol-l-phosphate (IP1) in the presence of LiCl. Carbachol, norepinephrine (NE) and high K+ increased accumulation of IP1 in slices from control rats. GABA was without effect on IP1 accumulation but potentiated the stimulation of PIP, hydrolysis by NE. NE-stimulated IP1 accumulation in slices from rats undergoing seizures was significantly reduced. GABA potentiation of the NE-stimulated hydrolysis was also reduced.

Filbert, M.G.; Phann, S.; Forster, J.; Ballough, G.P.; Cann, F.J.

1993-05-13

53

Phosphoinositide 3-kinases: A conserved family of signal transducers  

Microsoft Academic Search

Phosphoinositide 3-kinases (PI3Ks) generate lipids that are implicated in receptor-stimulated signalling and in the regulation of membrane traffic. Several distinct classes of PI3Ks have now been identified that have been conserved throughout eukaryotic evolution. Potential signalling pathways downstream of PI3Ks have been elucidated and PI3K function is now being characterised in several model organisms.

Bart Vanhaesebroeck; Sally J. Leevers; George Panayotou; Michael D. Waterfield

1997-01-01

54

Development of Phosphoinositide3 Kinase Pathway Inhibitors for Advanced Cancer  

Microsoft Academic Search

The phosphoinositide-3 kinase (PI3K) pathway plays a critical role in cancer cell growth and survival. PI3K is activated in\\u000a human cancers by elevated receptor tyrosine kinase activity, RAS mutation, as well as by mutation, amplification, and deletion\\u000a of genes encoding components of the pathway. Additionally, PI3K pathway activation plays an important role in acquired resistance\\u000a to both chemotherapy and targeted

James M. Cleary; Geoffrey I. Shapiro

2010-01-01

55

Phosphoinositides : key players in cell signalling, in time and space  

Microsoft Academic Search

Over the last few years, many reports have extended our knowledge of the inositol lipid metabolism and brought out some exciting information about the location, the variety and the role of phosphoinositides (PIs). Besides the so-called “canonical PI pathway” leading to the production of phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), the precursor of the intracellular second messengers inositol 1,4,5-trisphosphate and diacylglycerol (DAG), many

Bernard Payrastre; Karine Missy; Sylvie Giuriato; Stéphane Bodin; Monique Plantavid; Marie-Pierre Gratacap

2001-01-01

56

Targeting the phosphoinositide 3-kinase pathway in cancer  

Microsoft Academic Search

The phosphoinositide 3-kinase (PI3K) pathway is a key signal transduction system that links oncogenes and multiple receptor classes to many essential cellular functions, and is perhaps the most commonly activated signalling pathway in human cancer. This pathway therefore presents both an opportunity and a challenge for cancer therapy. Even as inhibitors that target PI3K isoforms and other major nodes in

Pixu Liu; Hailing Cheng; Thomas M. Roberts; Jean J. Zhao

2009-01-01

57

Essential Role of Phosphoinositide Metabolism in Synaptic Vesicle Recycling  

Microsoft Academic Search

Growing evidence suggests that phosphoinositides play an important role in membrane traffic. A polyphosphoinositide phosphatase, synaptojanin 1, was identified as a major presynaptic protein associated with endocytic coated intermediates. We report here that synaptojanin 1–deficient mice exhibit neurological defects and die shortly after birth. In neurons of mutant animals, PI(4,5)P2 levels are increased, and clathrin-coated vesicles accumulate in the cytomatrix-rich

Ottavio Cremona; Gilbert Di Paolo; Markus R Wenk; Anita Lüthi; Warren T Kim; Kohji Takei; Laurie Daniell; Yasuo Nemoto; Stephen B Shears; Richard A Flavell; David A McCormick; Pietro De Camilli

1999-01-01

58

Signalling through the lipid products of phosphoinositide-3OH kinase  

Microsoft Academic Search

When a stimulatory agonist molecule binds at the exterior of the cell membrane, a second messenger transduces the signal to the interior of the cell. Second messengers can be derived from phospholipids in the membrane by the action of the enzymes phospholipase C or phosphoinositide-3-OH kinase (PI(3)K). PI(3)K is a key player in many cellular responses, including the movement of

Alex Toker; Lewis C. Cantley

1997-01-01

59

Effects of soman-induced convulsions on phosphoinositide metabolism  

Microsoft Academic Search

Turnover of [3H]phosphoinositides (PI) was examined in brain slices from the hippocampus of rats undergoing soman-induced seizure activity.\\u000a Hydrolysis of PI was determined by measuring the accumulation of [3H]inositol-1-phosphate (IP1). Incubation of hippocampal slices in the presence of carbachol or norepinephrine (NE) increased PI hydrolysis. Stimulated\\u000a hydrolysis by NE, but not carbachol was significantly reduced in slices from soman-challenged rats

Margaret G. Filbert; Jeffry S. Forster; Sorabe Phann; Gerald P. H. Ballough

1998-01-01

60

Targeting phosphoinositide 3-kinase—Moving towards therapy  

Microsoft Academic Search

Phosphoinositide 3-kinases (PI3K) orchestrate cell responses including mitogenic signaling, cell survival and growth, metabolic control, vesicular trafficking, degranulation, cytoskeletal rearrangement and migration. Deregulation of the PI3K pathway occurs by activating mutations in growth factor receptors or the PIK3CA locus coding for PI3K?, by loss of function of the lipid phosphatase and tensin homolog deleted in chromosome ten (PTEN\\/MMAC\\/TEP1), by the

Romina Marone; Vladimir Cmiljanovic; Bernd Giese; Matthias P. Wymann

2008-01-01

61

Phosphoinositides: tiny lipids with giant impact on cell regulation.  

PubMed

Phosphoinositides (PIs) make up only a small fraction of cellular phospholipids, yet they control almost all aspects of a cell's life and death. These lipids gained tremendous research interest as plasma membrane signaling molecules when discovered in the 1970s and 1980s. Research in the last 15 years has added a wide range of biological processes regulated by PIs, turning these lipids into one of the most universal signaling entities in eukaryotic cells. PIs control organelle biology by regulating vesicular trafficking, but they also modulate lipid distribution and metabolism via their close relationship with lipid transfer proteins. PIs regulate ion channels, pumps, and transporters and control both endocytic and exocytic processes. The nuclear phosphoinositides have grown from being an epiphenomenon to a research area of its own. As expected from such pleiotropic regulators, derangements of phosphoinositide metabolism are responsible for a number of human diseases ranging from rare genetic disorders to the most common ones such as cancer, obesity, and diabetes. Moreover, it is increasingly evident that a number of infectious agents hijack the PI regulatory systems of host cells for their intracellular movements, replication, and assembly. As a result, PI converting enzymes began to be noticed by pharmaceutical companies as potential therapeutic targets. This review is an attempt to give an overview of this enormous research field focusing on major developments in diverse areas of basic science linked to cellular physiology and disease. PMID:23899561

Balla, Tamas

2013-07-01

62

Cloning and Characterization of a G Protein-Activated Human Phosphoinositide3 Kinase  

Microsoft Academic Search

Phosphoinositide-3 kinase activity is implicated in diverse cellular responses triggered by mammalian cell surface receptors and in the regulation of protein sorting in yeast. Receptors with intrinsic and associated tyrosine kinase activity recruit heterodimeric phosphoinositide-3 kinases that consist of p110 catalytic subunits and p85 adaptor molecules containing Src homology 2 (SH2) domains. A phosphoinositide-3 kinase isotype, p110gamma, was cloned and

Borislav Stoyanov; Stefano Volinia; Theodor Hanck; Ignacio Rubio; Michael Loubtchenkov; Daria Malek; Stefka Stoyanova; Bart Vanhaesebroeck; Ritu Dhand; Bernd Nurnberg; Peter Gierschik; Klaus Seedorf; J. Justin Hsuan; Michael D. Waterfield; Reinhard Wetzker

1995-01-01

63

Phosphoinositide 3-kinase enhancer (PIKE) in the brain: is it simply a phosphoinositide 3-kinase/Akt enhancer?  

PubMed Central

Since its discovery in 2000, phosphoinositide 3-kinase enhancer (PIKE) has been recognized as a class of GTPase that controls the enzymatic activities of phosphoinositide 3-kinase (PI3K) and Akt in the central nervous system (CNS). However, recent studies suggest that PIKEs are not only enhancers to PI3K/Akt but also modulators to other kinases including insulin receptor tyrosine kinase and focal adhesion kinases. Moreover, they regulate transcription factors such as signal transducer and activator of transcription and nuclear factor ?B. Indeed, PIKE proteins participate in multiple cellular processes including control of cell survival, brain development, memory formation, gene transcription, and metabolism. In this review, we have summarized the functions of PIKE proteins in CNS and discussed their potential implications in various neurological disorders. PMID:22499674

Chan, Chi Bun; Ye, Keqiang

2013-01-01

64

Stimulatory effects of maitotoxin on insulin release in insulinoma HIT cells: Role of calcium uptake and phosphoinositide breakdown  

SciTech Connect

In hamster insulinoma (HIT) cells, maitotoxin (MTX) induces a time-dependent and concentration-dependent release of insulin that requires the presence of extracellular calcium. The response is nearly completely blocked by cinnarizine and cadmium, but is not inhibited by the L-type calcium channel blocker nifedipine or by manganese. MTX induces 45Ca+ uptake in these cells in a dose-dependent mode, and the uptake is blocked with cinnarizine, nifedipine and cadmium, and is partially inhibited by manganese. MTX induces phosphoinositide breakdown in HIT cells, and the response is partially blocked by cadmium, but is not affected by nifedipine, cinnarizine or manganese. High concentrations of potassium ions also induce insulin release and calcium uptake in HIT cells. Both effects of potassium are blocked partially by nifedipine, cadmium and cinnarizine. High concentrations of potassium do not induce phosphoinositide breakdown in HIT cells. The results suggest that MTX-elicited release of insulin is attained by two mechanisms: (1) a nifedipine-sensitive action, which results from MTX-induced activation of L-type calcium channels, which can be mimicked with high potassium concentrations; and (2) a nifedipine-insensitive action, which may be initiated by the activation of phosphoinositide breakdown by MTX. Such an activation of phospholipase C would result in the formation of 1,4,5-inositol trisphosphate, a release of intracellular calcium and then release of insulin to the extracellular space. Cinnarizine is proposed to block both MTX-elicited mechanisms, the first by blockade of calcium channels and the second by blocking 1,4,5-inositol trisphosphate-induced release of internal calcium. Either mechanism alone appears capable of eliciting release of insulin.

Soergel, D.G.; Gusovsky, F.; Yasumoto, T.; Daly, J.W. (National Institute of Diabetes, Digestive and Kidney Diseases, Bethesda, MD (USA))

1990-12-01

65

VISUALIZIATION OF CELLULAR PHOSPHOINOSITIDE POOLS WITH GFP-FUSED PROTEIN-DOMAINS  

PubMed Central

This unit describes the method of following phosphoinositide dynamics in live cells. Inositol phospholipids have emerged as universal signaling molecules present in virtually every membrane of eukaryotic cells. Phosphoinositides are present only in tiny amounts compared to structural lipids but are metabolically very active as they are produced and degraded by the numerous inositide kinase and phosphatase enzymes. Phosphoinositides control the membrane-recruitment and activity of many protein signaling-complexes in specific membrane compartments and have been implicated in the regulation of a variety of signaling and trafficking pathways. It has been a challenge to develop methods that allow detection of phosphoinositides at the single cell level. The only available technique in live cell application is based on the use of the same protein domains selected by evolution to recognize cellular phosphoinositides. Some of these isolated protein modules when fused to fluorescent proteins can follow dynamic changes in phosphoinositides. While this technique can provide information on phosphoinositide dynamics in live cells with subcellular resolution and rapidly gained popularity, it also has several limitations that must be taken into account when interpreting the data. Here, we summarize the design and practical use of these constructs and also review important considerations for the interpretation of the data obtained by this technique. PMID:19283730

Balla, Tamas; Várnai, Péter

2011-01-01

66

Reduction of phosphoinositides and diacylglycerol levels in repeatedly dibutyltin-dilaurate-treated rat brain.  

PubMed

Oral administration of a single dose of dibutyltin dilaurate (DBTL, 80 mg/kg body wt.), 2 or 24 h after treatment, caused no significant change in the levels of diacylglycerol and phosphoinositides in rat cerebrum (forebrain), whereas daily administration of DBTL (40 or 80 mg/kg body wt.) for 3 days (24 h after the final treatment) decreased the levels of diacylglycerol and phosphoinositides (phosphatidylinositol, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate) in a concentration-dependent manner without influencing the levels of phosphatidylcholine in rat cerebrum. These studies indicate impairment of the phosphoinositide messenger system in rat cerebrum following repeated exposure to DBTL. PMID:1652800

Subramoniam, A; Husain, R; Seth, P K

1991-08-01

67

Phosphoinositide 3-kinase signalling in lung disease: leucocytes and beyond  

PubMed Central

The family of lipid kinases termed phosphoinositide-3-kinase (PI3K) is known to contribute at multiple levels to innate and adaptive immune responses, and is hence an attractive target for drug discovery in inflammatory and autoimmune disease, including respiratory diseases. The development of isoform-selective pharmacological inhibitors, targeted gene manipulation and short interfering RNA (siRNA) target validation have facilitated a better understanding of the role that each member of this family of kinases plays in the physiology and pathology of the respiratory system. In this review, we will evaluate the evidence for the roles of specific PI3K isoforms in the lung and airways, and discuss their potential as targets for novel drug therapies. PMID:17614878

Medina-Tato, David A; Ward, Stephen G; Watson, Malcolm L

2007-01-01

68

Activation of intracellular phosphoinositide signaling after a single 600 nanosecond electric pulse.  

PubMed

Exposure to nanosecond pulsed electrical fields (nsPEFs) results in a myriad of observable effects in mammalian cells. While these effects are often attributed to the direct permeabilization of both the plasma and organelle membranes, the underlying mechanism(s) are not well understood. We hypothesize that nsPEF-induced membrane disturbance will initiate complex intracellular lipid signaling pathways, which ultimately lead to the observed multifarious effects. In this article, we show activation of one of these pathways--phosphoinositide signaling cascade. Here we demonstrate that nsPEF initiates phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) hydrolysis or depletion from the plasma membrane, accumulation of inositol-1,4,5-trisphosphate (IP3) in the cytoplasm and increase of diacylglycerol (DAG) on the inner surface of the plasma membrane. All of these events are initiated by a single 16.2 kV/cm, 600 ns pulse exposure. To further this claim, we show that the nsPEF-induced activation mirrors the response of M1-acetylcholine Gq/11-coupled metabotropic receptor (hM1). This demonstration of PIP2 hydrolysis by nsPEF exposure is an important step toward understanding the mechanisms underlying this unique stimulus for activation of lipid signaling pathways and is critical for determining the potential for nsPEFs to modulate mammalian cell functions. PMID:23747521

Tolstykh, Gleb P; Beier, Hope T; Roth, Caleb C; Thompson, Gary L; Payne, Jason A; Kuipers, Marjorie A; Ibey, Bennett L

2013-12-01

69

Regulation of phosphoinositide hydrolysis and fibrinogen receptor expression by GTP regulatory protein(s) in isolated platelet membranes  

SciTech Connect

Thrombin through a GTP-dependent mechanism, stimulated fibrinogen binding and indogenous (/sup 3/H) inositol- and (/sup 32/P)-labeled phosphoinositide hydrolysis in membranes prepared from platelets. Incubation of isolated membranes with GTP enhanced both fibrinogen binding and phosphoinositide metabolism in a dose dependent fashion with half-maximal activation at approximately 50..mu..m and 25..mu..m respectively. Addition of thrombin (l..mu../ml) or ADP (10..mu..M) and GTP enhanced fibrinogen lending 10-fold compared to GTP alone. Inositol triphosphate formation was increased by 300 to 500% within 60 seconds after the addition of thrombin (l..mu../ml). Addition of thrombin plus GTP, however, did not lead to the formation of inositol on glycerol phosphoinositol. These data suggest that in a cell free system these processes are mediated by guanine nucleotide binding regulatory protein(s). Although a general role for GTP-binding proteins in receptor-regulated processes is currently a popular idea, this data is the most direct evidence to date for a role in platelets other than the well established mediation of adenylate cyclase activity.

Baldassare, J.J.; Fisher, G.

1986-05-01

70

Phosphoinositide kinases and the synthesis of polyphosphoinositides in higher plant cells  

NASA Technical Reports Server (NTRS)

Phosphoinositides are a family of inositol-containing phospholipids which are present in all eukaryotic cells. Although in most cells these lipids, with the exception of phosphatidylinositol, constitute only a very minor proportion of total cellular lipids, they have received immense attention by researchers in the past 15-20 years. This is due to the discovery that these lipids, rather than just having structural functions, play key roles in a wide range of important cellular processes. Much less is known about the plant phosphoinositides than about their mammalian counterparts. However, it has been established that a functional phosphoinositide system exists in plant cells and it is becoming increasingly clear that inositol-containing lipids are likely to play many important roles throughout the life of a plant. It is not our intention to give an exhaustive overview of all aspects of the field, but rather we focus on the phosphoinositide kinases responsible for the synthesis of all phosphorylated forms of phosphatidylinositol. Also, we mention some of the aspects of current phosphoinositide research which, in our opinion, are most likely to provide a suitable starting point for further research into the role of phosphoinositides in plants.

Drobak, B. K.; Dewey, R. E.; Boss, W. F.; Davies, E. (Principal Investigator)

1999-01-01

71

Crystal structure of the yeast Sac1: implications for its phosphoinositide phosphatase function  

PubMed Central

Sac family phosphoinositide (PI) phosphatases are an essential family of CX5R(T/S)-based enzymes, involved in numerous aspects of cellular function such as PI homeostasis, cellular signalling, and membrane trafficking. Genetic deletions of several Sac family members result in lethality in animal models and mutations of the Sac3 gene have been found in human hereditary diseases. In this study, we report the crystal structure of a founding member of this family, the Sac phosphatase domain of yeast Sac1. The 2.0 Å resolution structure shows that the Sac domain comprises of two closely packed sub-domains, a novel N-terminal sub-domain and the PI phosphatase catalytic sub-domain. The structure further shows a striking conformation of the catalytic P-loop and a large positively charged groove at the catalytic site. These findings suggest an unusual mechanism for its dephosphorylation function. Homology structural modeling of human Fig4/Sac3 allows the mapping of several disease-related mutations and provides a framework for the understanding of the molecular mechanisms of human diseases. PMID:20389282

Manford, Andrew; Xia, Tian; Saxena, Ajay Kumar; Stefan, Christopher; Hu, Fenghua; Emr, Scott D; Mao, Yuxin

2010-01-01

72

Phosphoinositide system-linked serotonin receptor subtypes and their pharmacological properties and clinical correlates.  

PubMed Central

Serotonergic neurotransmission represents a complex mechanism involving pre- and post-synaptic events and distinct 5-HT receptor subtypes. Serotonin (5-HT) receptors have been classified into several categories, and they are termed as 5-HT1, 5-HT2, 5-HT3, 5-HT4, 5-HT5, 5-HT6 and 5-HT7 type receptors. 5-HT1 receptors have been further subdivided into 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1E and 5-HT1F. 5-HT2 receptors have been divided into 5-HT2A, 5-HT2B and 5-HT2C receptors. All 5-HT2 receptor subtypes are linked to the multifunctional phosphoinositide (PI) signalling system. 5-HT3 receptors are considered ion-gated receptors and are also linked to the PI signalling system by an unknown mechanism. The 5-HT2A receptor subtype is the most widely studied of the 5-HT receptors in psychiatric disorders (for example, suicide, depression and schizophrenia) as well as in relation to the mechanism of action of antidepressant drugs. The roles of 5-HT2C and 5-HT3 receptors in psychiatric disorders are less clear. These 5-HT receptors also play an important role in alcoholism. It has been shown that 5-HT2A, 5-HT2C and 5-HT3 antagonists cause attenuation of alcohol intake in animals and humans. However, the exact mechanisms are unknown. The recent cloning of the cDNAs for 5-HT2A, 5-HT2C and 5-HT3 receptors provides the opportunity to explore the molecular mechanisms responsible for the alterations in these receptors during illness as well as pharmacotherapy. This review article will focus on the current research into the pharmacological properties, molecular biology, and clinical correlates of 5-HT2A, 5-HT2C and 5-HT3 receptors. PMID:7786883

Pandey, S C; Davis, J M; Pandey, G N

1995-01-01

73

Dual P2Y 12 receptor signaling in thrombin-stimulated platelets--involvement of phosphoinositide 3-kinase beta but not gamma isoform in Ca2+ mobilization and procoagulant activity.  

PubMed

During thrombus formation, thrombin, which is abundantly present at sites of vascular injury, activates platelets in part via autocrine-produced ADP. We investigated the signaling pathways by which thrombin and ADP in synergy induced platelet Ca(2+) elevation and procoagulant activity, and we monitored the consequences for the coagulation process. Even at high thrombin concentration, autocrine and added ADP enhanced and prolonged Ca(2+) depletion from internal stores via stimulation of the P2Y(12) receptors. This P2Y(12)-dependent effect was mediated via two distinct signaling pathways. The first is enhanced Ca(2+) mobilization by the inositol 1,4,5-trisphosphate receptors due to inhibition of protein kinase A. The second pathway concerns prolonged activation of phosphoinositide 3-kinase (PI3-K) and phospholipase C. Experiments with phosphoinositide 3-kinase isoform-selective inhibitors and p110gamma deficient platelets demonstrated that the phosphoinositide 3-kinase beta and not the phosphoinositide 3-kinase gamma isoform is responsible for the prolonged Ca(2+) response and for the subsequent increases in procoagulant activity and coagulation. Taken together, these results demonstrate a dual P2Y(12)-dependent signaling mechanism, which increases the platelet-activating effect of thrombin by prolongation of Ca(2+) elevation, thereby facilitating the coagulation process. PMID:18081863

van der Meijden, Paola E J; Schoenwaelder, Simone M; Feijge, Marion A H; Cosemans, Judith M E M; Munnix, Imke C A; Wetzker, Reinhard; Heller, Regine; Jackson, Shaun P; Heemskerk, Johan W M

2008-01-01

74

Phosphoinositides Differentially Regulate Protrudin Localization through the FYVE Domain*  

PubMed Central

Protrudin is a FYVE (Fab 1, YOTB, Vac 1, and EEA1) domain-containing protein involved in transport of neuronal cargoes and implicated in the onset of hereditary spastic paraplegia. Our image-based screening of the lipid binding domain library revealed novel plasma membrane localization of the FYVE domain of protrudin unlike canonical FYVE domains that are localized to early endosomes. The membrane binding study by surface plasmon resonance analysis showed that this FYVE domain preferentially binds phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2), phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P2), and phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3) unlike canonical FYVE domains that specifically bind phosphatidylinositol 3-phosphate (PtdIns(3)P). Furthermore, we found that these phosphoinositides (PtdInsP) differentially regulate shuttling of protrudin between endosomes and plasma membrane via its FYVE domain. Protrudin mutants with reduced PtdInsP-binding affinity failed to promote neurite outgrowth in primary cultured hippocampal neurons. These results suggest that novel PtdInsP selectivity of the protrudin-FYVE domain is critical for its cellular localization and its role in neurite outgrowth. PMID:23043110

Gil, Jung-Eun; Kim, Eui; Kim, Il-Shin; Ku, Bonsu; Park, Wei Sun; Oh, Byung-Ha; Ryu, Sung Ho; Cho, Wonhwa; Heo, Won Do

2012-01-01

75

Phosphoinositides, inositol phosphates, and phospholipase C in embryonic stem cells.  

PubMed

The stimulation of inositol phospholipid metabolism via phospholipase C (PLC) is an important signal transduction pathway in a wide variety of cell types. Activation of the pathway is associated with many aspects of cellular activity, including cell growth and differentiation. Activation of hormone-sensitive PLC results in the rapid breakdown of polyphosphoinositides to generate two second messengers: inositol trisphosphate and diacylglycerol. The water-soluble inositol trisphosphate is involved in the release of intracellular calcium from internal stores, whereas the lipophilic diacylglycerol is involved in protein kinase C activation. Inositol supplementation is essential for the in vitro growth of rabbit blastocysts, and studies have shown that the components of the signaling system are present in mouse and cattle embryos and in mouse embryonic stem (ES) cells. In ES cells, the signaling system appears to be constitutively active and essential for normal ES cell proliferation. Here, we describe in detail the materials required and some of techniques involved in studying the phosphoinositide signaling system in mouse ES cells. Furthermore, we describe methods of analyzing the effects of modulating the PtdIns signaling system on ES cell proliferation and the induction of apoptosis. PMID:16845989

Quinlan, Leo R

2006-01-01

76

Phosphoinositide regulation of inward rectifier potassium (Kir) channels  

PubMed Central

Inward rectifier potassium (Kir) channels are integral membrane proteins charged with a key role in establishing the resting membrane potential of excitable cells through selective control of the permeation of K+ ions across cell membranes. In conjunction with secondary anionic phospholipids, members of this family are directly regulated by phosphoinositides (PIPs) in the absence of other proteins or downstream signaling pathways. Different Kir isoforms display distinct specificities for the activating PIPs but all eukaryotic Kir channels are activated by PI(4,5)P2. On the other hand, the bacterial KirBac1.1 channel is inhibited by PIPs. Recent crystal structures of eukaryotic Kir channels in apo and lipid bound forms reveal one specific binding site per subunit, formed at the interface of N- and C-terminal domains, just beyond the transmembrane segments and clearly involving some of the key residues previously identified as controlling PI(4,5)P2 sensitivity. Computational, biochemical, and biophysical approaches have attempted to address the energetic determinants of PIP binding and selectivity among Kir channel isoforms, as well as the conformational changes that trigger channel gating. Here we review our current understanding of the molecular determinants of PIP regulation of Kir channel activity, including in context with other lipid modulators, and provide further discussion on the key questions that remain to be answered. PMID:24409153

Furst, Oliver; Mondou, Benoit; D'Avanzo, Nazzareno

2014-01-01

77

The Drosophila phosphoinositide 3-kinase Dp110 promotes cell growth.  

PubMed Central

Phosphoinositide 3-kinases (PI3Ks) have been identified in an evolutionarily diverse range of organisms, including mammals, Drosophila, yeast, plants and Dictyostelium. They are activated by a multitude of extracellular signals and implicated in mitogenesis, differentiation and cell survival, as well as in the control of the cytoskeleton and cell shape. Here we describe the molecular and functional analysis of Drosophila p110 (Dp110). A full-length Dp110 cDNA was isolated and found to encode a protein homologous throughout its length to the class I mammalian PI3Ks p110alpha and p110beta. Overexpression of Dp110 in wing or eye imaginal discs resulted in flies with enlarged wings or eyes respectively. In contrast, overexpression of Dp110 containing a mutation predicted to result in the loss of catalytic activity resulted in smaller wings and eyes. The alterations in wing size result from changes in both cell size and cell number, whereas in the eye only differences in cell size were detected. These data imply a role for Dp110 in growth control during Drosophila development and have implications for the function of class I PI3Ks in other organisms. Images PMID:8978685

Leevers, S J; Weinkove, D; MacDougall, L K; Hafen, E; Waterfield, M D

1996-01-01

78

Identification and Structural Characterization of a Legionella Phosphoinositide Phosphatase*  

PubMed Central

Bacterial pathogen Legionella pneumophila is the causative agent of Legionnaires' disease, which is associated with intracellular replication of the bacteria in macrophages of human innate immune system. Recent studies indicate that pathogenic bacteria can subvert host cell phosphoinositide (PI) metabolism by translocated virulence effectors. However, in which manner Legionella actively exploits PI lipids to benefit its infection is not well characterized. Here we report that L. pneumophila encodes an effector protein, named SidP, that functions as a PI-3-phosphatase specifically hydrolyzing PI(3)P and PI(3,5)P2 in vitro. This activity of SidP rescues the growth phenotype of a yeast strain defective in PI(3)P phosphatase activity. Crystal structure of SidP orthologue from Legionella longbeachae reveals that this unique PI-3-phosphatase is composed of three distinct domains: a large catalytic domain, an appendage domain that is inserted into the N-terminal portion of the catalytic domain, and a C-terminal ?-helical domain. SidP has a small catalytic pocket that presumably provides substrate specificity by limiting the accessibility of bulky PIs with multiple phosphate groups. Together, our identification of a unique family of Legionella PI phosphatases highlights a common scheme of exploiting host PI lipids in many intracellular bacterial pathogen infections. PMID:23843460

Toulabi, Leila; Wu, Xiaochun; Cheng, Yanshu; Mao, Yuxin

2013-01-01

79

NuMA interacts with phosphoinositides and links the mitotic spindle with the plasma membrane.  

PubMed

The positioning and the elongation of the mitotic spindle must be carefully regulated. In human cells, the evolutionary conserved proteins LGN/G?i1-3 anchor the coiled-coil protein NuMA and dynein to the cell cortex during metaphase, thus ensuring proper spindle positioning. The mechanisms governing cortical localization of NuMA and dynein during anaphase remain more elusive. Here, we report that LGN/G?i1-3 are dispensable for NuMA-dependent cortical dynein enrichment during anaphase. We further establish that NuMA is excluded from the equatorial region of the cell cortex in a manner that depends on the centralspindlin components CYK4 and MKLP1. Importantly, we reveal that NuMA can directly associate with PtdInsP (PIP) and PtdInsP2 (PIP2) phosphoinositides in vitro. Furthermore, chemical or enzymatic depletion of PIP/PIP2 prevents NuMA cortical localization during mitosis, and conversely, increasing PIP2 levels augments mitotic cortical NuMA. Overall, our study uncovers a novel function for plasma membrane phospholipids in governing cortical NuMA distribution and thus the proper execution of mitosis. PMID:24996901

Kotak, Sachin; Busso, Coralie; Gönczy, Pierre

2014-08-18

80

New experimental trends for phosphoinositides research on ion transporter/channel regulation.  

PubMed

Phosphoinositides(4,5)-bisphosphates [PI(4,5)P2] critically controls membrane excitability, the disruption of which leads to pathophysiological states. PI(4,5)P2 plays a primary role in regulating the conduction and gating properties of ion channels/transporters, through electrostatic and hydrophobic interactions that allow direct associations. In recent years, the development of many molecular tools have brought deep insights into the mechanisms underlying PI(4,5)P2-mediated regulation. This review summarizes the methods currently available to manipulate the cell membrane PI(4,5)P2 level including pharmacological interventions as well as newly designed molecular tools. We concisely introduce materials and experimental designs suitable for the study of PI(4,5)P2-mediated regulation of ion-conducting molecules, in order to assist researchers who are interested in this area. It is our further hope that the knowledge introduced in this review will help to promote our understanding about the pathology of diseases such as cardiac arrhythmias, bipolar disorders, and Alzheimer's disease which are somehow associated with a disruption of PI(4,5)P2 metabolism. PMID:25367262

Mori, Masayuki X; Inoue, Ryuji

2014-11-20

81

The Sac domain-containing phosphoinositide phosphatases: structure, function, and disease  

PubMed Central

Phosphoinositides (PIs) have long been known to have an essential role in cell physiology. Their intracellular localization and concentration must be tightly regulated for their proper function. This spatial and temporal regulation is achieved by a large number of PI kinases and phosphatases that are present throughout eukaryotic species. One family of these enzymes contains a conserved PI phosphatase domain termed Sac. Although the Sac domain is homologous among different Sac domain-containing proteins, all appear to exhibit varied substrate specificity and subcellular localization. Dysfunctions in several members of this family are implicated in a range of human diseases such as cardiac hypertrophy, bipolar disorder, Down’s syndrome, Charcot-Marie-Tooth disease (CMT) and Amyotrophic Lateral Sclerosis (ALS). In plant, several Sac domain-containing proteins have been implicated in the stress response, chloroplast function and polarized secretion. In this review, we focus on recent findings in the family of Sac domain-containing PI phosphatases in yeast, mammal and plant, including the structural analysis into the mechanism of enzymatic activity, cellular functions, and their roles in disease pathophysiology. PMID:24860601

HSU, FoSheng; MAO, Yuxin

2014-01-01

82

Stimulation of phosphoinositide hydrolysis by thrombin in embryonic chick heart cells  

SciTech Connect

The authors have shown that muscarinic receptor stimulation results in phosphoinositide (PI) hydrolysis in dissociated embryonic chick heart cells, with maximal stimulation produced by 1 mM carbachol. In an effort to identify other stimulators of PI turnover, they studied the effects of thrombin in primary cultures of 13-day embryonic chick heart cells. Thrombin increased PI hydrolysis, as measured by the accumulation of (/sup 3/H)inositol 1-phosphate in the presence of 10 mM LiCl. Stimulation occurred in a dose-dependent manner with half-maximal stimulation obtained at 0.07 U/ml and maximal stimulation at 0.3 U/ml thrombin. The effects of maximal doses of carbachol and thrombin were additive, suggesting different mechanisms of action or different populations of cells. Effects of both hormones were inhibited by pretreatment of the monolayer cultures with 4..beta..-phorbol 12 ..beta..-myristate 13..cap alpha..-acetate. The simultaneous addition of hirudin with thrombin blocked thrombin-stimulated PI turnover. An active-site-blocked derivative of thrombin was ineffective in stimulating IP formation. These data indicate a proteolytic action of thrombin in stimulation of the PI response. To their knowledge, these are the first data suggesting that there may be thrombin receptors or effects of thrombin on heart cells.

Jones, L.G.; Brown, J.H.

1986-03-05

83

Drosophila neuroligin 1 regulates synaptic growth and function in response to activity and phosphoinositide-3-kinase.  

PubMed

Neuroligins are postsynaptic neural cell adhesion molecules that mediate synaptic maturation and function in vertebrates and invertebrates, but their mechanisms of action and regulation are not well understood. At the Drosophila larval neuromuscular junction (NMJ), previous analysis demonstrated a requirement for Drosophila neuroligin 1 (dnlg1) in synaptic growth and maturation. The goal of the present study was to better understand the effects and mechanisms of loss-of-function and overexpression of dnlg1 on synapse size and function, and to identify signaling pathways that control dnlg1 expression. Consistent with reduced synapse size, evoked excitatory junctional currents (EJCs) were diminished in dnlg1 mutants but displayed normal Ca(2+) sensitivity and short-term plasticity. However, postsynaptic function was also perturbed, in that glutamate receptor staining and the distribution of amplitudes of miniature excitatory junctional currents (mEJCs) were abnormal in mutants. All the above phenotypes were rescued by a genomic transgene. Overexpression of dnlg1 in muscle resulted in synaptic overgrowth, but reduced the amplitudes of EJCs and mEJCs. Overgrowth and reduced EJC amplitude required Drosophila neurexin 1 (dnrx1) function, suggesting that increased DNlg1/DNrx1 signaling attenuates synaptic transmission and regulates growth through a retrograde mechanism. In contrast, reduced mEJC amplitude was independent of dnrx1. Synaptic overgrowth, triggered by neuronal hyperactivity, absence of the E3 ubiquitin ligase highwire, and increased phosphoinositide-3-kinase (PI3K) signaling in motor neurons reduced synaptic DNlg1 levels. Likewise, postsynaptic attenuation of PI3K, which increases synaptic strength, was associated with reduced DNlg1 levels. These observations suggest that activity and PI3K signaling pathways modulate growth and synaptic transmission through dnlg1-dependent mechanisms. PMID:22954894

Mozer, Brian A; Sandstrom, David J

2012-11-01

84

A Drosophila neuroligin regulates synaptic growth and function in response to activity and phosphoinositide-3-kinase  

PubMed Central

Neuroligins are postsynaptic neural cell adhesion molecules that mediate synaptic maturation and function in vertebrates and invertebrates, but their mechanisms of action and regulation are not well understood. At the Drosophila larval neuromuscular junction (NMJ), previous analysis demonstrated a requirement for Drosophila neuroligin 1 (dnlg1) in synaptic growth and maturation. The goal of the present study was to better understand the effects and mechanisms of loss-of-function and overexpression of dnlg1 on synapse size and function, and to identify signaling pathways that control dnlg1 expression. Consistent with a reduced synapse size, evoked excitatory junctional currents (EJCs) were diminished in dnlg1 mutants but displayed normal Ca2+ sensitivity and short-term plasticity. However, postsynaptic function was also perturbed, in that glutamate receptor staining and the distribution of amplitudes of miniature excitatory junctional currents (mEJCs) were abnormal in mutants. All the above phenotypes were rescued by a genomic transgene. Overexpression of dnlg1 in muscle resulted in synaptic overgrowth, but reduced the amplitudes of EJCs and mEJCs. Overgrowth and reduced EJC amplitude required dnrx1 function, suggesting that increased dnlg1/dnrx1 signaling attenuates synaptic transmission and regulates growth through a retrograde mechanism. In contrast, reduced mEJC amplitude was independent of dnrx1. Synaptic overgrowth, triggered by neuronal hyperactivity, absence of the E3 ubiquitin ligase highwire, and increased phosphoinositide-3-kinase (PI3K) signaling in motor neurons reduced synaptic DNlg1 levels. Likewise, postsynaptic attenuation of PI3K, which increases synaptic strength, was associated with reduced DNlg1 levels. These observations suggest that activity and PI3K signaling pathways modulate growth and synaptic transmission through dnlg1-dependent mechanisms. PMID:22954894

Mozer, Brian A.; Sandstrom, David J.

2012-01-01

85

Nicotine Promotes Late Endothelial Progenitor Cells Functional Activity in a PI 3-Kinase-Dependent Manner.  

PubMed

Recent studies have shown that endothelial progenitor cells (EPCs) participated in angiogenic effects of nicotine and nicotine dose dependently increased the functional activity of early EPCs. The effects of nicotine on late EPCs remain to be determined. Therefore, we investigated whether nicotine had influences on the functional activity of late EPCs. Late EPCs were isolated from human umbilical cord blood and characterized. Late EPCs of 3-5 passages were treated for 32 h with either vehicle or nicotine. The proliferative, migratory, and in vitro vasculogenesis activities of late EPCs were assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, modified Boyden chamber assay, and in matrigel, respectively. Late EPCs adhesion assay was performed by replating cells on fibronectin-coated dishes, and then adherent cells were counted. Nicotine enhanced proliferative, migratory, adhesive, and in vitro vasculogenesis capacities of late EPCs. These effects were significantly reduced in the presence of phosphatidylinositol (PI) 3-kinase inhibitor. PMID:24817640

Yu, Min; Li, Zhi; Shu, Zhouwu; Liu, Qian; Sun, Jing; Tan, Xuerui

2014-11-01

86

Human Rhinovirus 1B Exposure Induces Phosphatidylinositol 3-Kinase-dependent Airway Inflammation in Mice  

Microsoft Academic Search

Rationale: Infection with rhinovirus (RV) triggers exacerbations of asthma and chronic obstructive lung disease. Objectives: We sought to develop a mouse model of RV employing RV1B, a minor group serotype that binds to the low-density lipopro- tein receptor. Methods: C57BL\\/6 mice were inoculated intranasally with RV1B, replication-deficient ultraviolet (UV)-irradiated RV1B, or RV39, a major group virus. Measurements and Main Results:

Dawn C. Newcomb; Umadevi S. Sajjan; Deepti R. Nagarkar; Qiong Wang; Suparna Nanua; Ying Zhou; Christina L. McHenry; Kenneth T. Hennrick; Wan C. Tsai; J. Kelley Bentley; Nicholas W. Lukacs; Sebastian L. Johnston; Marc B. Hershenson

87

Genome-Wide Analysis of the Phosphoinositide Kinome from Two Ciliates Reveals Novel Evolutionary Links for Phosphoinositide Kinases in Eukaryotic Cells  

PubMed Central

Background The complexity of phosphoinositide signaling in higher eukaryotes is partly due to expansion of specific families and types of phosphoinositide kinases (PIKs) that can generate all phosphoinositides via multiple routes. This is particularly evident in the PI3Ks and PIPKs, and it is considered an evolutionary trait associated with metazoan diversification. Yet, there are limited comprehensive studies on the PIK repertoire of free living unicellular organisms. Methodology/Principal Findings We undertook a genome-wide analysis of putative PIK genes in two free living ciliated cells, Tetrahymena and Paramecium. The Tetrahymena thermophila and Paramecium tetraurelia genomes were probed with representative kinases from all families and types. Putative homologs were verified by EST, microarray and deep RNA sequencing database searches and further characterized for domain structure, catalytic efficiency, expression patterns and phylogenetic relationships. In total, we identified and characterized 22 genes in the Tetrahymena thermophila genome and 62 highly homologues genes in Paramecium tetraurelia suggesting a tight evolutionary conservation in the ciliate lineage. Comparison to the kinome of fungi reveals a significant expansion of PIK genes in ciliates. Conclusions/Significance Our study highlights four important aspects concerning ciliate and other unicellular PIKs. First, ciliate-specific expansion of PI4KIII-like genes. Second, presence of class I PI3Ks which, at least in Tetrahymena, are associated with a metazoan-type machinery for PIP3 signaling. Third, expansion of divergent PIPK enzymes such as the recently described type IV transmembrane PIPKs. Fourth, presence of possible type II PIPKs and presumably inactive PIKs (hence, pseudo-PIKs) not previously described. Taken together, our results provide a solid framework for future investigation of the roles of PIKs in ciliates and indicate that novel functions and novel regulatory pathways of phosphoinositides may be more widespread than previously thought in unicellular organisms. PMID:24244373

Leondaritis, George; Siokos, John; Skaripa, Irini; Galanopoulou, Dia

2013-01-01

88

Phosphoinositide 3-kinase at the crossroad between endocytosis and signaling of cytokine receptors.  

PubMed

Class I phosphoinositide 3-kinase (PI3K) is a lipid kinase playing key roles in many signaling pathways regulating cell survival and growth. Besides its important role in signal transduction, PI3K is also involved in actin and membrane reorganization such as protrusion, adhesion, phagocytosis and macropinocytosis. Receptor-mediated endocytosis is initiated by plasma membrane reorganization creating buds that then mature to small vesicles. Whereas most of endocytic mechanisms involve actin polymerization, PI3K requirement has not been clearly investigated. Our study identifies class I PI3K as a key player in clathrin-independent endocytosis of the interleukin 2 receptor (IL-2R) in contrast to the clathrin-dependent entry of transferrin (Tf). IL-2R is a cytokine receptor, inducing several signaling cascades such as PI3K, that are essential for the immune response. We have shown previously that IL-2R can be internalized with or without IL-2 and this process requires dynamin, actin and their regulators cortactin, N-WASP, Rac1 and the kinases Pak. Our recent work reveals that PI3K regulates Rac1 during IL-2R uptake in two ways: via its catalytic activity (p110) and via its regulatory factor (p85). Indeed, the catalytic activity of PI3K is required for both constitutive and IL-2 induced uptake of cytokine receptors, in lymphocytes as well as in epithelial cells. Interestingly, Vav2, a Rac1 GTPase exchange factor (GEF) induced upon PI3K activation, is specifically involved and recruited during IL-2R uptake. The second action of PI3K is via its regulatory subunit, p85, which binds activated Rac1 and IL-2R; this interaction being enhanced upon IL-2 treatment. Thus, PI3K regulates both the activation of Rac1 and its recruitment during IL-2R endocytosis. Finally, our results identify a link between cytokine receptors signaling and clathrin-independent endocytosis. PMID:23986799

Basquin, Cyril; Sauvonnet, Nathalie

2013-07-01

89

Phosphoinositide-3 kinase-Akt pathway controls cellular entry of Ebola virus.  

PubMed

The phosphoinositide-3 kinase (PI3K) pathway regulates diverse cellular activities related to cell growth, migration, survival, and vesicular trafficking. It is known that Ebola virus requires endocytosis to establish an infection. However, the cellular signals that mediate this uptake were unknown for Ebola virus as well as many other viruses. Here, the involvement of PI3K in Ebola virus entry was studied. A novel and critical role of the PI3K signaling pathway was demonstrated in cell entry of Zaire Ebola virus (ZEBOV). Inhibitors of PI3K and Akt significantly reduced infection by ZEBOV at an early step during the replication cycle. Furthermore, phosphorylation of Akt-1 was induced shortly after exposure of cells to radiation-inactivated ZEBOV, indicating that the virus actively induces the PI3K pathway and that replication was not required for this induction. Subsequent use of pseudotyped Ebola virus and/or Ebola virus-like particles, in a novel virus entry assay, provided evidence that activity of PI3K/Akt is required at the virus entry step. Class 1A PI3Ks appear to play a predominant role in regulating ZEBOV entry, and Rac1 is a key downstream effector in this regulatory cascade. Confocal imaging of fluorescently labeled ZEBOV indicated that inhibition of PI3K, Akt, or Rac1 disrupted normal uptake of virus particles into cells and resulted in aberrant accumulation of virus into a cytosolic compartment that was non-permissive for membrane fusion. We conclude that PI3K-mediated signaling plays an important role in regulating vesicular trafficking of ZEBOV necessary for cell entry. Disruption of this signaling leads to inappropriate trafficking within the cell and a block in steps leading to membrane fusion. These findings extend our current understanding of Ebola virus entry mechanism and may help in devising useful new strategies for treatment of Ebola virus infection. PMID:18769720

Saeed, Mohammad F; Kolokoltsov, Andrey A; Freiberg, Alexander N; Holbrook, Michael R; Davey, Robert A

2008-01-01

90

IN VITRO ALUMINUM INHIBITION OF BRAIN PHOSPHOINOSITIDE METABOLISM:COMPARISON OF NEONATAL AND ADULTS RATS  

EPA Science Inventory

Recent evidence indicates that the neurotoxic metal aluminum interferes with the phosphoinositide second messenger system in adult rats both in vitro and in vivo. e have examined the age-related effects of aluminum chloride (AlCl3) on receptor-stimulated inositol phosphate (IP) a...

91

Abscisic Acid-Induced Phosphoinositide Turnover in Guard Cell Protoplasts of Vicia faba.  

PubMed Central

Guard cell protoplasts of Vicia faba treated with 10 [mu]M (+)abscisic acid (ABA) in the light exhibited a 20% decrease in diameter within 1.5 h, from 24.1 to 19.6 [mu]m. Within 10 s of administration of ABA, a 90% increase in levels of inositol 1,4,5-trisphosphate was observed, provided that cells were treated with Li+, an inhibitor of inositol phosphatase activity, prior to incubation. Concomitantly, levels of 32P-labeled phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-phosphate decreased 20% compared to levels in control cells; levels of label in the membrane lipids phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol did not change significantly in response to ABA treatment. These results show that phosphoinositide turnover is activated in response to ABA in guard cells. We conclude that phosphoinositide signaling is likely to be a step in the biochemical cascade that couples ABA to guard cell shrinking and stomatal closure. PMID:12226236

Lee, Y.; Choi, Y. B.; Suh, S.; Lee, J.; Assmann, S. M.; Joe, C. O.; Kelleher, J. F.; Crain, R. C.

1996-01-01

92

Atg18 phosphoregulation controls organellar dynamics by modulating its phosphoinositide-binding activity  

PubMed Central

The PROPPIN family member Atg18 is a phosphoinositide-binding protein that is composed of a seven ?-propeller motif and is part of the conserved autophagy machinery. Here, we report that the Atg18 phosphorylation in the loops in the propellar structure of blade 6 and blade 7 decreases its binding affinity to phosphatidylinositol 3,5-bisphosphate in the yeast Pichia pastoris. Dephosphorylation of Atg18 was necessary for its association with the vacuolar membrane and caused septation of the vacuole. Upon or after dissociation from the vacuolar membrane, Atg18 was rephosphorylated, and the vacuoles fused and formed a single rounded structure. Vacuolar dynamics were regulated according to osmotic changes, oxidative stresses, and nutrient conditions inducing micropexophagy via modulation of Atg18 phosphorylation. This study reveals how the phosphoinositide-binding activity of the PROPPIN family protein Atg18 is regulated at the membrane association domain and highlights the importance of such phosphoregulation in coordinated intracellular reorganization. PMID:23940117

Tamura, Naoki; Oku, Masahide; Ito, Moemi; Noda, Nobuo N.; Inagaki, Fuyuhiko

2013-01-01

93

Phosphoinositide3 Kinase-Akt Pathway Controls Cellular Entry of Ebola Virus  

Microsoft Academic Search

The phosphoinositide-3 kinase (PI3K) pathway regulates diverse cellular activities related to cell growth, migration, survival, and vesicular trafficking. It is known that Ebola virus requires endocytosis to establish an infection. However, the cellular signals that mediate this uptake were unknown for Ebola virus as well as many other viruses. Here, the involvement of PI3K in Ebola virus entry was studied.

Mohammad F. Saeed; Andrey A. Kolokoltsov; Alexander N. Freiberg; Michael R. Holbrook; Robert A. Davey

2008-01-01

94

Cholinergic activation of phosphoinositide signaling is impaired in Alzheimer's disease brain  

Microsoft Academic Search

The function of the phosphoinositide signal transduction system was compared in membranes from Alzheimer's disease (AD) and control postmortem brain. [3H]Phosphatidylinositol hydrolysis was concentration-dependently stimulated by GTP[S] and this was 40% lower than controls in AD prefrontal cortical membranes. Carbachol induced a response greater than that of GTP[S] alone, and this response was impaired in AD by 45%. Differential analysis

Richard S. Jope; Ling Song; Richard E. Powers

1997-01-01

95

Regulation of Breast Cancer Cell Chemotaxis by the Phosphoinositide 3Kinase p1101  

Microsoft Academic Search

Class IA phosphoinositide 3-kinases (PI3Ks) regulate many cellular processes downstream of tyrosine kinases and Ras. Despite a clear impli- cation of PI3K in cancer, little is known about the distribution of the different PI3K isoforms in malignant cells. We screened a large panel of tissues and cell lines for expression of class IA PI3Ks, and document a ubiquitous expression of

Carol Sawyer; Justin Sturge; Dorothy C. Bennett; Michael J. O'Hare; William E. Allen; Jennifer Bain; Gareth E. Jones; Bart Vanhaesebroeck

2003-01-01

96

Heterogeneity of cholinergic muscarinic receptors coupled to phosphoinositide metabolism in immature rat brain  

Microsoft Academic Search

The effects of muscarinic agonists and antagonists on phosphoinositide (PtdIns) metabolism were examined in the cerebral cortex and brainstem of 7-day-old rats, in order to evaluate the role of muscarinic receptor subtypes in this process. Additionally, comparative experiments were performed in cortices from adult animals. Accumulation of [3H]inositol phosphates ([3H]InsPs) in [3H]inositol pre-labeled brain slices was taken as an index

Stefano M. Candura; Marcello Tonini; Paola Baiardi; Luigi Manzo; Lucio G. Costa

1995-01-01

97

Class IA Phosphoinositide 3Kinase Regulates Heart Size and Physiological Cardiac Hypertrophy  

Microsoft Academic Search

Class IA phosphoinositide 3-kinases (PI3Ks) are activated by growth factor receptors, and they regulate, among other processes, cell growth and organ size. Studies using transgenic mice overexpressing constitutively active and dominant negative forms of the p110 catalytic subunit of class IA PI3K have implicated the role of this enzyme in regulating heart size and physiological cardiac hypertrophy. To further understand

Ji Luo; Julie R. McMullen; Cassandra L. Sobkiw; Li Zhang; Adam L. Dorfman; Megan C. Sherwood; M. Nicole Logsdon; James W. Horner; Ronald A. DePinho; Seigo Izumo; Lewis C. Cantley

2005-01-01

98

Interaction of PDK1 with Phosphoinositides Is Essential for Neuronal Differentiation but Dispensable for Neuronal Survival  

PubMed Central

3-Phosphoinositide-dependent protein kinase 1 (PDK1) operates in cells in response to phosphoinositide 3-kinase activation and phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3] production by activating a number of AGC kinases, including protein kinase B (PKB)/Akt. Both PDK1 and PKB contain pleckstrin homology (PH) domains that interact with the PtdIns(3,4,5)P3 second messenger. Disrupting the interaction of the PDK1 PH domain with phosphoinositides by expressing the PDK1 K465E knock-in mutation resulted in mice with reduced PKB activation. We explored the physiological consequences of this biochemical lesion in the central nervous system. The PDK1 knock-in mice displayed a reduced brain size due to a reduction in neuronal cell size rather than cell number. Reduced BDNF-induced phosphorylation of PKB at Thr308, the PDK1 site, was observed in the mutant neurons, which was not rate limiting for the phosphorylation of those PKB substrates governing neuronal survival and apoptosis, such as FOXO1 or glycogen synthase kinase 3 (GSK3). Accordingly, the integrity of the PDK1 PH domain was not essential to support the survival of different embryonic neuronal populations analyzed. In contrast, PKB-mediated phosphorylation of PRAS40 and TSC2, allowing optimal mTORC1 activation and brain-specific kinase (BRSK) protein synthesis, was markedly reduced in the mutant mice, leading to impaired neuronal growth and differentiation. PMID:23275438

Zurashvili, Tinatin; Cordon-Barris, Lluis; Ruiz-Babot, Gerard; Zhou, Xiangyu; Lizcano, Jose M.; Gomez, Nestor; Gimenez-Llort, Lydia

2013-01-01

99

Sequential breakdown of 3-phosphorylated phosphoinositides is essential for the completion of macropinocytosis  

PubMed Central

Macropinocytosis is a highly conserved endocytic process by which extracellular fluid and solutes are internalized into cells. Macropinocytosis starts with the formation of membrane ruffles at the plasma membrane and ends with their closure. The transient and sequential emergence of phosphoinositides PI(3,4,5)P3 and PI(3,4)P2 in the membrane ruffles is essential for macropinocytosis. By making use of information in the Caenorhabditis elegans mutants defective in fluid-phase endocytosis, we found that mammalian phosphoinositide phosphatase MTMR6 that dephosphorylates PI(3)P to PI, and its binding partner MTMR9, are required for macropinocytosis. INPP4B, which dephosphorylates PI(3,4)P2 to PI(3)P, was also found to be essential for macropinocytosis. These phosphatases operate after the formation of membrane ruffles to complete macropinocytosis. Finally, we showed that KCa3.1, a Ca2+-activated K+ channel that is activated by PI(3)P, is required for macropinocytosis. We propose that the sequential breakdown of PI(3,4,5)P3 ? PI(3,4)P2 ? PI(3)P ? PI controls macropinocytosis through specific effectors of the intermediate phosphoinositides. PMID:24591580

Maekawa, Masashi; Terasaka, Shimpei; Mochizuki, Yasuhiro; Kawai, Katsuhisa; Ikeda, Yuka; Araki, Nobukazu; Skolnik, Edward Y.; Taguchi, Tomohiko; Arai, Hiroyuki

2014-01-01

100

The Phosphoinositide Kinase PIKfyve/Fab1p Regulates Terminal Lysosome Maturation in Caenorhabditis elegans  

PubMed Central

Membrane dynamics is necessary for cell homeostasis and signal transduction and is in part regulated by phosphoinositides. Pikfyve/Fab1p is a phosphoinositide kinase that phosphorylates phosphatidylinositol 3-monophosphate into phosphatidylinositol-3,5-bisphosphate [PtdIns(3,5)P2] and is implicated in membrane homeostasis in yeast and in mammalian cells. These two phosphoinositides are substrates of myotubularin phosphatases found mutated in neuromuscular diseases. We studied the roles of phosphatidylinositol phosphate kinase 3 (PPK-3), the orthologue of PIKfyve/Fab1p, in a multicellular organism, Caenorhabditis elegans. Complete loss of ppk-3 function induces developmental defects characterized by embryonic lethality, whereas partial loss of function leads to growth retardation. At the cellular level, ppk-3 mutants display a striking enlargement of vacuoles positive for lysosome-associated membrane protein 1 in different tissues. In the intestine, RAB-7–positive late endosomes are also enlarged. Membranes of the enlarged lysosomes originate at least in part from smaller lysosomes, and functional and genetic analyses show that the terminal maturation of lysosomes is defective. Protein degradation is not affected in the hypomorphic ppk-3 mutant and is thus uncoupled from membrane retrieval. We measured the level of PtdIns(3,5)P2 and showed that its production is impaired in this mutant. This work strongly suggests that the main function of PPK-3 is to mediate membrane retrieval from matured lysosomes through regulation of PtdIns(3,5)P2. PMID:16801682

Nicot, Anne-Sophie; Fares, Hanna; Payrastre, Bernard; Chisholm, Andrew D.; Labouesse, Michel

2006-01-01

101

ERK and phosphoinositide 3-kinase temporally coordinate different modes of actin-based motility during embryonic wound healing.  

PubMed

Embryonic wound healing provides a perfect example of efficient recovery of tissue integrity and homeostasis, which is vital for survival. Tissue movement in embryonic wound healing requires two functionally distinct actin structures: a contractile actomyosin cable and actin protrusions at the leading edge. Here, we report that the discrete formation and function of these two structures is achieved by the temporal segregation of two intracellular upstream signals and distinct downstream targets. The sequential activation of ERK and phosphoinositide 3-kinase (PI3K) signalling divides Xenopus embryonic wound healing into two phases. In the first phase, activated ERK suppresses PI3K activity, and is responsible for the activation of Rho and myosin-2, which drives actomyosin cable formation and constriction. The second phase is dominated by restored PI3K signalling, which enhances Rac and Cdc42 activity, leading to the formation of actin protrusions that drive migration and zippering. These findings reveal a new mechanism for coordinating different modes of actin-based motility in a complex tissue setting, namely embryonic wound healing. PMID:23986484

Li, Jingjing; Zhang, Siwei; Soto, Ximena; Woolner, Sarah; Amaya, Enrique

2013-11-01

102

TbFRP, a novel FYVE-domain containing phosphoinositide-binding Ras-like GTPase from trypanosomes.  

PubMed

Ras-like small GTPases are regulatory proteins that control multiple aspects of cellular function, and are particularly prevalent in vesicular transport. A proportion of GTPase paralogs appear restricted to certain eukaryote lineages, suggesting roles specific to a restricted lineage, and hence potentially reflecting adaptation to individual lifestyles or ecological niche. Here we describe the role of a GTPase, TbFRP, a FYVE domain N-terminally fused to a Ras-like GTPase, originally identified in Trypanosoma brucei. As FYVE-domains specifically bind phosphoinositol 3-phosphate (PI3P), which associates with endosomes, we suggest that TbFRP may unite phosphoinositide and small G protein endosomal signaling in trypanosomatids. TbFRP orthologs are present throughout the Euglenazoa suggesting that FRP has functions throughout the group. We show that the FYVE domain of TbFRP is functional in PI3P-dependent membrane targeting and localizes at the endosomal region. Further, while TbFRP is apparently non-essential, knockdown and immunochemical evidence indicates that TbFRP is rapidly cleaved upon synthesis, releasing the GTPase and FYVE-domains. Finally, TbFRP expression at both mRNA and protein levels is cell density-dependent. Together, these data suggest that TbFRP is an endocytic GTPase with a highly unusual mechanism of action that involves proteolysis of the nascent protein and membrane targeting via PI3P. PMID:23220323

Adung'a, Vincent O; Field, Mark C

2013-03-01

103

CCR7 ligands control basal T cell motility within lymph node slices in a phosphoinositide 3-kinase- independent manner  

PubMed Central

The molecular mechanisms responsible for the sustained basal motility of T cells within lymph nodes (LNs) remain elusive. To study T cell motility in a LN environment, we have developed a new experimental system based on slices of LNs that allows the assessment of T cell trafficking after adoptive transfer or direct addition of T cells to the slice. Using this experimental system, we show that T cell motility is highly sensitive to pertussis toxin and strongly depends on CCR7 and its ligands. Our results also demonstrate that, despite its established role in myeloid cell locomotion, phosphoinositide 3–kinase (PI3K) activity does not contribute to the exploratory behavior of the T lymphocytes within LN slices. Likewise, although PI3K activation is detectable in chemokine-treated T cells, PI3K plays only a minor role in T cell polarization and migration in vitro. Collectively, our results suggest that the common amplification system that, in other cells, facilitates large phosphatidylinositol 3,4,5-trisphosphate increases at the plasma membrane is absent in T cells. We conclude that T cell motility within LNs is not an intrinsic property of T lymphocytes but is driven in a PI3K-independent manner by the lymphoid chemokine-rich environment. PMID:17485513

Asperti-Boursin, Francois; Real, Eliana; Bismuth, Georges; Trautmann, Alain; Donnadieu, Emmanuel

2007-01-01

104

CCR7 ligands control basal T cell motility within lymph node slices in a phosphoinositide 3-kinase-independent manner.  

PubMed

The molecular mechanisms responsible for the sustained basal motility of T cells within lymph nodes (LNs) remain elusive. To study T cell motility in a LN environment, we have developed a new experimental system based on slices of LNs that allows the assessment of T cell trafficking after adoptive transfer or direct addition of T cells to the slice. Using this experimental system, we show that T cell motility is highly sensitive to pertussis toxin and strongly depends on CCR7 and its ligands. Our results also demonstrate that, despite its established role in myeloid cell locomotion, phosphoinositide 3-kinase (PI3K) activity does not contribute to the exploratory behavior of the T lymphocytes within LN slices. Likewise, although PI3K activation is detectable in chemokine-treated T cells, PI3K plays only a minor role in T cell polarization and migration in vitro. Collectively, our results suggest that the common amplification system that, in other cells, facilitates large phosphatidylinositol 3,4,5-trisphosphate increases at the plasma membrane is absent in T cells. We conclude that T cell motility within LNs is not an intrinsic property of T lymphocytes but is driven in a PI3K-independent manner by the lymphoid chemokine-rich environment. PMID:17485513

Asperti-Boursin, François; Real, Eliana; Bismuth, Georges; Trautmann, Alain; Donnadieu, Emmanuel

2007-05-14

105

Assessing the subcellular distribution of oncogenic phosphoinositide 3-kinase using microinjection into live cells  

PubMed Central

Oncogenic mutations in PIK3CA lead to an increase in intrinsic phosphoinositide kinase activity, but it is thought that increased access of PI3K? (phosphoinositide 3-kinase ?) to its PM (plasma membrane) localized substrate is also required for increased levels of downstream PIP3/Akt [phosphoinositide-3,4,5-trisphosphate/also called PKB (protein kinase B)] signalling. We have studied the subcellular localization of wild-type and the two most common oncogenic mutants of PI3K? in cells maintained in growth media, and starved or stimulated cells using a novel method in which PI3K? is pre-formed as a 1:1 p110?:p85? complex in vitro then introduced into live cells by microinjection. Oncogenic E545K and H1047R mutants did not constitutively interact with membrane lipids in vitro or in cells maintained in 10% (v/v) FBS. Following stimulation of RTKs (receptor tyrosine kinases), microinjected PI3K? was recruited to the PM, but oncogenic forms of PI3K? were not recruited to the PM to a greater extent and did not reside at the PM longer than the wild-type PI3K?. Instead, the E545K mutant specifically bound activated Cdc42 in vitro and microinjection of E545K was associated with the formation of cellular protrusions, providing some preliminary evidence that changes in protein–protein interactions may play a role in the oncogenicity of the E545K mutant in addition to the well-known changes in lipid kinase activity. PMID:24597785

Layton, Meredith J.; Rynkiewicz, Natalie K.; Ivetac, Ivan; Horan, Kristy A.; Mitchell, Christina A.; Phillips, Wayne A.

2014-01-01

106

Spatial Localization of m-Calpain to the Plasma Membrane by Phosphoinositide Biphosphate Binding during Epidermal Growth Factor Receptor-Mediated Activation†  

PubMed Central

Calpain activity is required for de-adhesion of the cell body and rear to enable productive locomotion of adherent cells during wound repair and tumor invasion. Growth factors activate m-calpain (calpain 2, CAPN2) via ERK/mitogen-activated protein kinases, but only when these kinases are localized to the plasma membrane. We thus hypothesized that m-calpain is activated by epidermal growth factor (EGF) only when it is juxtaposed to the plasma membrane secondary to specific docking. Osmotic disruption of NR6 fibroblasts expressing the EGF receptor demonstrated m-calpain being complexed with the substratum-adherent membrane with this increasing in an EGF-dependent manner. m-Calpain colocalized with phosphoinositide biphosphate (PIP2) with exogenous phospholipase C removal of phosphoinositides, specifically, PI(4,5)P2 but not PI(4)P1 or PIP3, releasing the bound m-calpain. Downregulation of phosphoinositide production by 1-butanol resulted in diminished PIP2 in the plasma membrane and eliminated EGF-induced calpain activation. This PIP2-binding capacity resided in domain III of calpain, which presents a putative C2-like domain. This active conformation of this domain appears to be partially masked in the holoenzyme as both activation of m-calpain by phosphorylation at serine 50 and expression of constitutively active phosphorylation mimic glutamic acid-increased m-calpain binding to the membrane, consistent with blockade of this cascade diminishing membrane association. Importantly, we found that m-calpain was enriched toward the rear of locomoting cells, which was more pronounced in the plasma membrane footprints; EGF further enhanced this enrichment, in line with earlier reports of loss of PIP2 in lamellipodia of motile cells. These data support a model of m-calpain binding to PIP2 concurrent with and likely to enable ERK activation and provides a mechanism by which cell de-adhesion is directed to the cell body and tail as phospholipase C-? hydrolyzes PIP2 in the protruding lamellipodia. PMID:16809781

Shao, Hanshuang; Chou, Jeff; Baty, Catherine J.; Burke, Nancy A.; Watkins, Simon C.; Stolz, Donna Beer; Wells, Alan

2006-01-01

107

Estradiol attenuates prolactin secretion and phosphoinositide hydrolysis in MMQ cells.  

PubMed

We previously isolated a clonal cell line, designated MMQ, which only secretes prolactin (PRL) and whose secretory process is nonresponsive to thyrotropin releasing hormone (TRH) and angiotensin II (AII). In the present study, we injected MMQ cells into rats to determine whether the tumor cells would become responsive to secretagogues when subsequently propagated in vitro. We also investigated what effects in vivo administration of 17 beta-estradiol would have on secretagogue-induced PRL release and on intracellular biochemical mechanisms in these cells. MMQ cells were implanted subcutaneously in the backs of female rats. One group was injected with 100 micrograms polyestradiol phosphate (PEP) every 5 days, a second with saline. The inoculants grew into solid tumors within 3 weeks. The day after the tumors were removed and enzymatically dispersed, the cells, now designated MMQt cells, were perifused in vitro. Basal PRL released by MMQt cells was approximately 1 ng/min/10(7) cells and perifusions with 100 nM TRH or AII for 5 min significantly increased PRL release above baseline (integrated areas: 1.8 +/- 0.4 and 5.2 +/- 1.3 ng/10(7) cell, respectively; P less than 0.01). Two ng/ml maitotoxin (MTX), a calcium channel activator, increased PRL release (38.2 +/- 6.7 ng/10(7) cells; P less than 0.01). In PEP-treated perifused MMQt cells, basal in vitro PRL release was not different from that observed in the control group, but the responses to TRH, AII and MTX were greatly attenuated (TRH: 0.6 +/- 0.1, AII: 1.3 +/- 0.2 and MTX: 9.2 +/- 2.5 ng/10(7) cells).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2573549

Kubota, T; Login, I S; Judd, A M; Kuan, S I; MacLeod, R M

1989-09-01

108

The phosphoinositide-3-phosphatase MTMR2 associates with MTMR13, a membrane-associated pseudophosphatase also mutated in type 4B Charcot-Marie-Tooth disease.  

PubMed

Charcot-Marie-Tooth disease type 4B (CMT4B) is a severe, demyelinating peripheral neuropathy characterized by distinctive, focally folded myelin sheaths. CMT4B is caused by recessively inherited mutations in either myotubularin-related 2 (MTMR2) or MTMR13 (also called SET-binding factor 2). MTMR2 encodes a member of the myotubularin family of phosphoinositide-3-phosphatases, which dephosphorylate phosphatidylinositol 3-phosphate (PI(3)P) and bisphosphate PI(3,5)P2. MTMR13 encodes a large, uncharacterized member of the myotubularin family. The MTMR13 phosphatase domain is catalytically inactive because the essential Cys and Arg residues are absent. Given the genetic association of both MTMR2 and MTMR13 with CMT4B, we investigated the biochemical relationship between these two proteins. We found that the endogenous MTMR2 and MTMR13 proteins are associated in human embryonic kidney 293 cells. MTMR2-MTMR13 association is mediated by coiled-coil sequences present in each protein. We also examined the cellular localization of MTMR2 and MTMR13 using fluorescence microscopy and subcellular fractionation. We found that (i) MTMR13 is a predominantly membrane-associated protein; (ii) MTMR2 and MTMR13 cofractionate in both a light membrane fraction and a cytosolic fraction; and (iii) MTMR13 membrane association is mediated by the segment of the protein which contains the pseudophosphatase domain. This work, which describes the first cellular or biochemical investigation of the MTMR13 pseudophosphatase protein, suggests that MTMR13 functions in association with MTMR2. Loss of MTMR13 function in CMT4B2 patients may lead to alterations in MTMR2 function and subsequent alterations in 3-phosphoinositide signaling. Such a mechanism would explain the strikingly similar phenotypes of patients with recessive mutations in either MTMR2 or MTMR13. PMID:15998640

Robinson, Fred L; Dixon, Jack E

2005-09-01

109

Phosphoinositides: Minor Lipids Make a Major Impact on Photoreceptor Cell Functions  

PubMed Central

Activation of the phosphoinositide (PI) cycle generates the second messengers that control various aspects of cellular signaling. We have previously shown that two PI cycle enzymes, type II phosphatidylinositol 5-phosphate 4-kinase (PIPK II?) and phosphoinositide 3-kinase (PI3K), are activated through light stimulation. In our earlier studies, we measured enzyme activities, instead of directly measuring the products, due to lack of sensitive analytical techniques. Cells have very low levels of PIs, compared to other lipids, so special techniques and sensitive analytical instruments are necessary for their identification and quantification. There are also other considerations, such as different responses in different cell types, which may complicate quantification of PIs. For example, although light activated PIPK II?, there was no increase in PI-4,5-P2 measured by liquid chromatography–mass spectrometry (LC/MS) This discrepancy is due to the heterogeneous nature of the retina, which is composed of various cell types. In this study, we examined PI generation in situ using immunohistochemistry with specific PI antibodies. PIs were generated in specific retinal cell layers, suggesting that analyzing PIs from the total retina by LC/MS underscores the significance. This suggests that PI-specific antibodies are useful tools to study the cell-specific regulation of PIs in the retina. PMID:24964953

Rajala, Raju V. S.; Rajala, Ammaji; Morris, Andrew J.; Anderson, Robert E.

2014-01-01

110

The role of phosphoinositides and inositol phosphates in plant cell signaling.  

PubMed

Work over the recent years has greatly expanded our understanding of the specific molecules involved in plant phosphoinositide signaling. Physiological approaches, combined with analytical techniques and genetic mutants have provided tools to understand how individual genes function in this pathway. Several key differences between plants and animals have become apparent. This chapter will highlight the key areas where major differences between plants and animals occur. In particular, phospholipase C and levels of phosphatidylinositol phosphates differ between plants and animals, and may influence how inositol second messengers form and function in plants. Whether inositol 1,4,5-trisphosphate and/or inositol hexakisphosphate (InsP6) function as second messengers in plants is discussed. Recent data on potential, novel roles of InsP6 in plants is considered, along with the existence of a unique InsP6 synthesis pathway. Lastly, the complexity of myo-inositol synthesis in plants is discussed in reference to synthesis of phosphoinositides and impact on plant growth and development. PMID:23775694

Gillaspy, Glenda E

2013-01-01

111

Yunis-Varón syndrome is caused by mutations in FIG4, encoding a phosphoinositide phosphatase.  

PubMed

Yunis-Varón syndrome (YVS) is an autosomal-recessive disorder with cleidocranial dysplasia, digital anomalies, and severe neurological involvement. Enlarged vacuoles are found in neurons, muscle, and cartilage. By whole-exome sequencing, we identified frameshift and missense mutations of FIG4 in affected individuals from three unrelated families. FIG4 encodes a phosphoinositide phosphatase required for regulation of PI(3,5)P(2) levels, and thus endosomal trafficking and autophagy. In a functional assay, both missense substitutions failed to correct the vacuolar phenotype of Fig4-null mouse fibroblasts. Homozygous Fig4-null mice exhibit features of YVS, including neurodegeneration and enlarged vacuoles in neurons. We demonstrate that Fig4-null mice also have small skeletons with reduced trabecular bone volume and cortical thickness and that cultured osteoblasts accumulate large vacuoles. Our findings demonstrate that homozygosity or compound heterozygosity for null mutations of FIG4 is responsible for YVS, the most severe known human phenotype caused by defective phosphoinositide metabolism. In contrast, in Charcot-Marie-Tooth disease type 4J (also caused by FIG4 mutations), one of the FIG4 alleles is hypomorphic and disease is limited to the peripheral nervous system. This genotype-phenotype correlation demonstrates that absence of FIG4 activity leads to central nervous system dysfunction and extensive skeletal anomalies. Our results describe a role for PI(3,5)P(2) signaling in skeletal development and maintenance. PMID:23623387

Campeau, Philippe M; Lenk, Guy M; Lu, James T; Bae, Yangjin; Burrage, Lindsay; Turnpenny, Peter; Román Corona-Rivera, Jorge; Morandi, Lucia; Mora, Marina; Reutter, Heiko; Vulto-van Silfhout, Anneke T; Faivre, Laurence; Haan, Eric; Gibbs, Richard A; Meisler, Miriam H; Lee, Brendan H

2013-05-01

112

The Phosphoinositide 3-Kinase Regulatory Subunit p85  Can Exert Tumor Suppressor Properties through Negative Regulation of Growth Factor Signaling  

E-print Network

Phosphoinositide 3-kinase (PI3K) plays a critical role in tumorigenesis, and the PI3K p85 regulatory subunit exerts both positive and negative effects on signaling. Expression of Pik3r1, the gene encoding p85, is decreased ...

Taniguchi, Cullen M.

113

Phosphoinositide signaling.  

PubMed

"All things flow and change…even in the stillest matter there is unseen flux and movement." Attributed to Heraclitus (530-470 BC), from The Story of Philosophy by Will Durant. Heraclitus, a Greek philosopher, was thinking on a much larger scale than molecular signaling; however, his visionary comments are an important reminder for those studying signaling today. Even in unstimulated cells, signaling pathways are in constant metabolic flux and provide basal signals that travel throughout the organism. In addition, negatively charged phospholipids, such as the polyphosphorylated inositol phospholipids, provide a circuit board of on/off switches for attracting or repelling proteins that define the membranes of the cell. This template of charged phospholipids is sensitive to discrete changes and metabolic fluxes-e.g., in pH and cations-which contribute to the oscillating signals in the cell. The inherent complexities of a constantly fluctuating system make understanding how plants integrate and process signals challenging. In this review we discuss one aspect of lipid signaling: the inositol family of negatively charged phospholipids and their functions as molecular sensors and regulators of metabolic flux in plants. PMID:22404474

Boss, Wendy F; Im, Yang Ju

2012-01-01

114

Salicylic acid modulates levels of phosphoinositide dependent-phospholipase C substrates and products to remodel the Arabidopsis suspension cell transcriptome  

PubMed Central

Basal phosphoinositide-dependent phospholipase C (PI-PLC) activity controls gene expression in Arabidopsis suspension cells and seedlings. PI-PLC catalyzes the production of phosphorylated inositol and diacylglycerol (DAG) from phosphoinositides. It is not known how PI-PLC regulates the transcriptome although the action of DAG-kinase (DGK) on DAG immediately downstream from PI-PLC is responsible for some of the regulation. We previously established a list of genes whose expression is affected in the presence of PI-PLC inhibitors. Here this list of genes was used as a signature in similarity searches of curated plant hormone response transcriptome data. The strongest correlations obtained with the inhibited PI-PLC signature were with salicylic acid (SA) treatments. We confirm here that in Arabidopsis suspension cells SA treatment leads to an increase in phosphoinositides, then demonstrate that SA leads to a significant 20% decrease in phosphatidic acid, indicative of a decrease in PI-PLC products. Previous sets of microarray data were re-assessed. The SA response of one set of genes was dependent on phosphoinositides. Alterations in the levels of a second set of genes, mostly SA-repressed genes, could be related to decreases in PI-PLC products that occur in response to SA action. Together, the two groups of genes comprise at least 40% of all SA-responsive genes. Overall these two groups of genes are distinct in the functional categories of the proteins they encode, their promoter cis-elements and their regulation by DGK or phospholipase D. SA-regulated genes dependent on phosphoinositides are typical SA response genes while those with an SA response that is possibly dependent on PI-PLC products are less SA-specific. We propose a model in which SA inhibits PI-PLC activity and alters levels of PI-PLC products and substrates, thereby regulating gene expression divergently.

Ruelland, Eric; Pokotylo, Igor; Djafi, Nabila; Cantrel, Catherine; Repellin, Anne; Zachowski, Alain

2014-01-01

115

Wortmannin, a widely used phosphoinositide 3-kinase inhibitor, also potently inhibits mammalian polo-like kinase.  

PubMed

Polo-like kinases (PLKs) play critical roles throughout mitosis. Here, we report that wortmannin, which was previously thought to be a highly selective inhibitor of phosphoinositide (PI) 3-kinases, is a potent inhibitor of mammalian PLK1. Observation of the wortmannin-PLK1 interaction was enabled by a tetramethylrhodamine-wortmannin conjugate (AX7503) that permits rapid detection of PLK1 activity and expression in complex proteomes. Importantly, we show that wortmannin inhibits PLK1 activity in an in vitro kinase assay with an IC(50) of 24 nM and when incubated with intact cells. Taken together, our results indicate that, at the concentrations of wortmannin commonly used to inhibit PI 3-kinases, PLK1 is also significantly inhibited. PMID:15664519

Liu, Yongsheng; Shreder, Kevin R; Gai, Wenzhi; Corral, Sergio; Ferris, Douglas K; Rosenblum, Jonathan S

2005-01-01

116

The conserved phosphoinositide 3-kinase pathway determines heart size in mice  

PubMed Central

Phosphoinositide 3-kinase (PI3K) has been shown to regulate cell and organ size in Drosophila, but the role of PI3K in vertebrates in vivo is not well understood. To examine the role of PI3K in intact mammalian tissue, we have created and characterized transgenic mice expressing constitutively active or dominant-negative mutants of PI3K in the heart. Cardiac- specific expression of constitutively active PI3K resulted in mice with larger hearts, while dominant-negative PI3K resulted in mice with smaller hearts. The increase or decrease in heart size was associated with comparable increase or decrease in myocyte size. Cardiomyopathic changes, such as myocyte necrosis, apoptosis, interstitial fibrosis or contractile dysfunction, were not observed in either of the transgenic mice. Thus, the PI3K pathway is necessary and sufficient to promote organ growth in mammals. PMID:10835352

Shioi, Tetsuo; Kang, Peter M.; Douglas, Pamela S.; Hampe, James; Yballe, Claudine M.; Lawitts, Joel; Cantley, Lewis C.; Izumo, Seigo

2000-01-01

117

8-(Tosylamino)quinoline inhibits tumour progression through targeting phosphoinositide-3-kinase/Akt pathway.  

PubMed

We examined whether 8-(tosylamino)quinoline (8-TQ), a structural analogue of BAY 11-7082, is able to modulate various tumourigenic responses using various in vitro and in vivo experimental conditions. 8-TQ exhibited the strongest suppressive activity on the proliferation of C6, A431, HeLa and MDA-MB-231 cells with IC550 values ranging from 10 to 30 microM. According to the analysis of level of active caspase-3, and morphologies of C6, HeLa and MDA-MB-231 cells, it was revealed that 8-TQ is able to induce apoptosis. Furthermore, this compound strongly diminished the invasion of MDA-MB-231 cells, the migration of HeLa cells, and the new generation of blood vessels under non-toxic conditions. Reduction of the phospho-form levels of intracellular signalling enzymes by 8-TQ strongly indicated that molecular signalling machineries composed of phosphoinositide 3-kinase (PI3K)/phosphoinositide-dependent kinase-1 (PDK1)/Akt and extracellular-signal-regulated kinase (ERK) could be targeted by 8-TQ treatment. Indeed, the specific inhibitors (LY294002 and U0126) of PI3K/PDK1/Akt and ERK showed similar anti-cancer properties to 8-TQ. Finally, 8-TQ intraperitoneally injected suppressed the increase of tumour volume up to 40% compared to vehicle-treated control. Taken together, our results clearly suggest that 8-TQ might have applications as a novel anti-cancer drug or may be served as a lead compound to be further optimized. PMID:23469688

Jung, Yongwoo; Yi, Young-Su; Yoo, Dae Sung; Kim, Ji Hye; Yang, Woo Seok; Lee, Jongsung; Park, Kye Won; Kweon, Dae-Hyuk; Hong, Sungyoul; Cho, Jae Youl

2013-02-01

118

A new approach to measuring phosphoinositides in cells by mass spectrometry.  

PubMed

The phosphoinositide family of phospholipids, defined here as PtdIns, PtdIns3P, PtdIns4P, PtdIns5P, PtdIns(3,4)P2, PtdIns(3,5)P2, PtdIns(4,5)P2 and PtdIns(3,4,5)P3, play pivotal roles in organising the location and activity of many different proteins acting on biological membranes, including those involved in vesicle and protein trafficking through the endolysosomal system and receptor signal transduction at the plasma membrane. Accurate measurement of the cellular levels of these lipids, particularly the more highly phosphorylated species, is hampered by their high polarity and low cellular concentrations. Recently, much progress has been made in using mass spectrometry to measure many different lipid classes in parallel, an approach generally referred to as 'lipidomics'. Unfortunately, the acidic nature of highly phosphorylated phosphoinositides makes them difficult to measure using these methods, because they yield low levels of useful ions; this is particularly the case with PtdIns(3,4,5)P3. We have solved some of these problems by methylating the phosphate groups of these lipids with TMS-diazomethane and describe a simple, integrated approach to measuring PtdIns, PtdInsP, PtdInsP2 and PtdInsP3 classes of lipids, in parallel with other phospholipid species, in cell and tissue extracts. This methodology is sensitive, accurate and robust, and also yields fatty-acyl compositions, suggesting it can be used to further our understanding of both the normal and pathophysiological roles of these important lipids. PMID:24120934

Kielkowska, Anna; Niewczas, Izabella; Anderson, Karen E; Durrant, Tom N; Clark, Jonathan; Stephens, Len R; Hawkins, Phillip T

2014-01-01

119

Characterization of phosphoinositide-specific phospholipase C in rat colonocyte membranes.  

PubMed Central

The phosphoinositide signal transduction pathway mediates important processes in intestinal physiology, yet the key enzyme, phosphoinositide-specific phospholipase C (PI-PLC), is not well-characterized in the colon. PI-PLC activity was examined in rat colonic membranes using exogenous [3H]phosphatidylinositol 4,5-bisphosphate (PIP2) as substrate, and beta-glycerophosphate to suppress degradation of substrate or product. The activity of membrane PI-PLC increased 6-fold with the addition of alamethicin, and a further 2-3-fold enhancement was observed with 10 microM guanosine 5'-[gamma-thio]triphosphate (GTP[S]), suggesting the involvement of G-protein(s). The effect of GTP[S] appeared to be specific, as up to 100 microM adenosine 5'-[gamma-thio]-triphosphate failed to stimulate PI-PLC activity, and guanosine 5'-[beta-thio]diphosphate inhibited activity. The response of membrane PI-PLC to Ca2+ was biphasic, while > 0.5 mM Mg2+ was inhibitory with or without GTP[S]. Comparable total PI-PLC activities and responses to GTP[S] and Ca2+ were observed in purified brush-border and basolateral membranes. Western immunoblots probed with monoclonal antibodies to PLC isoenzymes PLC-beta 1, -gamma 1 and -delta 1 demonstrated that these antipodal plasma membranes contain predominantly the PLC-delta 1 isoform, with small amounts of PLC-gamma 1 present but no detectable PLC-beta 1. PLC-gamma 1 was the major isoform detected in cytosol. Images Figure 5 PMID:8389128

Bolt, M J; Bissonnette, B M; Wali, R K; Hartmann, S C; Brasitus, T A; Sitrin, M D

1993-01-01

120

Phosphoinositide-signaling is one component of a robust plant defense response  

PubMed Central

The phosphoinositide pathway and inositol-1,4,5-triphosphate (InsP3) have been implicated in plant responses to many abiotic stresses; however, their role in response to biotic stress is not well characterized. In the current study, we show that both basal defense and systemic acquired resistance responses are affected in transgenic plants constitutively expressing the human type I inositol polyphosphate 5-phosphatase (InsP 5-ptase) which have greatly reduced InsP3 levels. Flagellin induced Ca2+-release as well as the expressions of some flg22 responsive genes were attenuated in the InsP 5-ptase plants. Furthermore, the InsP 5-ptase plants were more susceptible to virulent and avirulent strains of Pseudomonas syringae pv. tomato (Pst) DC3000. The InsP 5-ptase plants had lower basal salicylic acid (SA) levels and the induction of SAR in systemic leaves was reduced and delayed. Reciprocal exudate experiments showed that although the InsP 5-ptase plants produced equally effective molecules that could trigger PR-1 gene expression in wild type plants, exudates collected from either wild type or InsP 5-ptase plants triggered less PR-1 gene expression in InsP 5-ptase plants. Additionally, expression profiles indicated that several defense genes including PR-1, PR-2, PR-5, and AIG1 were basally down regulated in the InsP 5-ptase plants compared with wild type. Upon pathogen attack, expression of these genes was either not induced or showed delayed induction in systemic leaves. Our study shows that phosphoinositide signaling is one component of the plant defense network and is involved in both basal and systemic responses. The dampening of InsP3-mediated signaling affects Ca2+ release, modulates defense gene expression and compromises plant defense responses. PMID:24966862

Hung, Chiu-Yueh; Aspesi Jr, Peter; Hunter, Melissa R.; Lomax, Aaron W.; Perera, Imara Y.

2014-01-01

121

A Ras Inhibitor Tilts the Balance between Rac and Rho and Blocks Phosphatidylinositol 3-Kinase-Dependent Glioblastoma Cell Migration  

Microsoft Academic Search

Glioblastoma multiforme are highly aggressive tumors for which no adequate treatment has yet been developed. Glioblastoma multiforme show large amounts of active Ras, considered an appropriate target for directed therapy. Here, we show that the Ras inhibitor S-trans, trans-farnesyl thiosalicylic acid (FTS) can avert the transformation of human glioblastoma multiforme cells by inhibiting both their migration and their anchorage-independent proliferation.

Liat Goldberg; Yoel Kloog

2006-01-01

122

PI3-kinase dependent activation of apoptotic machinery occurs on commitment of epidermal keratinocytes to terminal differentiation  

PubMed Central

We have investigated the earliest events in commitment of human epidermal keratinocytes to terminal differentiation. Phosphorylated Akt and caspase activation were detected in cells exiting the basal layer of the epidermis. Activation of Akt by retroviral transduction of primary cultures of human keratinocytes resulted in an increase in abortive clones founded by transit amplifying cells, while inhibition of the upstream kinase, PI3 kinase, inhibited suspension-induced terminal differentiation. Caspase inhibition also blocked differentiation, the primary mediator being caspase 8. Caspase activation was initiated by 2 hours in suspension, preceding the onset of expression of the terminal differentiation marker involucrin by several hours. Incubation of suspended cells with fibronectin or inhibition of PI3 kinase prevented caspase induction. At 2 hours in suspension, keratinocytes that had committed to terminal differentiation had increased side scatter, were 7AAD positive and annexin V negative; they exhibited loss of mitochondrial membrane potential and increased cardiolipin oxidation, but with no increase in reactive oxygen species. These properties indicate that the onset of terminal differentiation, while regulated by PI3 kinase and caspases, is not a classical apoptotic process. PMID:18766172

Janes, Sam M.; Ofstad, Tyler A.; Campbell, Douglas H.; Eddaoudi, Ayad; Warnes, Gary; Davies, Derek; Watt, Fiona M.

2008-01-01

123

Definition of the binding mode of a new class of phosphoinositide 3-kinase ?-selective inhibitors using in vitro mutagenesis of non-conserved amino acids and kinetic analysis  

PubMed Central

The binding mechanism of a new class of lipid-competitive, ATP non-competitive, p110? isoform-selective PI3K (phosphoinositide 3-kinase) inhibitors has been elucidated. Using the novel technique of isoform reciprocal mutagenesis of non-conserved amino acids in the p110? and p110? isoforms, we have identified three unique binding mechanisms for the p110?-selective inhibitors PIK-75, A-66S and J-32. Each of the inhibitor’s p110?-isoform-selective binding was found to be due to interactions with different amino acids within p110. The PIK-75 interaction bound the non-conserved region 2 amino acid p110? Ser773, A-66S bound the region 1 non-conserved amino acid p110? Gln859, and J-32 binding had an indirect interaction with Lys776 and Ile771. The isoform reciprocal mutagenesis technique is shown to be an important analytical tool for the rational design of isoform-selective inhibitors. PMID:22502592

Zheng, Zhaohua; Amran, Syazwani I.; Zhu, Jiuxiang; Schmidt-Kittler, Oleg; Kinzler, Kenneth W.; Vogelstein, Bert; Shepherd, Peter R.; Thompson, Philip E.; Jennings, Ian G.

2012-01-01

124

Growth hormone increases phosphoinositide turnover in rat adipocytes that are sensitive to the insulin-like action of the hormone  

Microsoft Academic Search

The effect of pituitary human growth hormone (hGH) on the 32P-labelling of phosphoinositides and phosphatidic acid was studied in noradrenaline-stimulated rat adipocytes which were either responsive or non-responsive to the antilipolytic (insulin-like) effect of hGH. In cells responsive to the insulin-like effect of hGH, hormone treatment resulted in a marked increase of the 32P-labelling of phosphatidic acid and phosphatidyl inositol

Hans Eriksson; Roger Sundler; Jakob Donnér

1990-01-01

125

Lead optimization of a dihydropyrrolopyrimidine inhibitor against phosphoinositide 3-kinase (PI3K) to improve the phenol glucuronic acid conjugation.  

PubMed

Our lead compound for a phosphoinositide 3-kinase (PI3K) inhibitor (1) was metabolically unstable because of rapid glucuronidation of the phenol moiety. Based on structure-activity relationship (SAR) information and a FlexSIS docking simulation score, aminopyrimidine was identified as a bioisostere of phenol. An X-ray structure study revealed a hydrogen bonding pattern of aminopyrimidine derivatives. Finally, aminopyrimidine derivatives 33 showed strong tumor growth inhibition against a KPL-4 breast cancer xenograft model in vivo. PMID:23265889

Kawada, Hatsuo; Ebiike, Hirosato; Tsukazaki, Masao; Nakamura, Mitsuaki; Morikami, Kenji; Yoshinari, Kiyoshi; Yoshida, Miyuki; Ogawa, Kotaro; Shimma, Nobuo; Tsukuda, Takuo; Ohwada, Jun

2013-02-01

126

The role of 3-phosphoinositide-dependent protein kinase 1 in activating AGC kinases defined in embryonic stem cells  

Microsoft Academic Search

Background: Protein kinase B (PKB), and the p70 and p90 ribosomal S6 kinases (p70 S6 kinase and p90 Rsk, respectively), are activated by phosphorylation of two residues, one in the ‘T-loop’ of the kinase domain and, the other, in the hydrophobic motif carboxy terminal to the kinase domain. The 3-phosphoinositide-dependent protein kinase 1 (PDK1) activates many AGC kinases in vitro

Michayla R. Williams; J. Simon C. Arthur; Anudharan Balendran; Jeroen van der Kaay; Valeria Poli; Philip Cohen; Dario R. Alessi

2000-01-01

127

Effects of the Metabotropic Glutamate Receptor Antagonist MCPG on Phosphoinositide Turnover and Synaptic Plasticity in Visual Cortex  

Microsoft Academic Search

The neurotransmitter glutamate, in addition to activating ligand- gated ion channels, also stimulates phosphoinositide (PI) hydro- lysis in neurons by activating a group of G-protein-coupled metabotropic glutamate receptors (mGluRs). A role for mGluRs in synaptic plasticity originally was hypothesized based on the ob- servation that the developmental decline in glutamate-stimulated PI turnover is well correlated with the decline in experience-

Kimberly M. Huber; Nathaniel B. Sawtell; Mark F. Bear

1998-01-01

128

Direct metabolic regulation of ?-catenin activity by the p85? regulatory subunit of phosphoinositide 3OH kinase  

Microsoft Academic Search

Class IA phosphoinositide 3-OH kinases (PI3K) are lipid kinases composed of catalytic and regulatory subunits. These lipid kinases can regulate the metabolic stability and signaling activity of ?-catenin, a central component of the E-cadherin\\/catenin cell–cell adhesion complex, and of the Wnt signaling pathway. This regulation occurs at the level of glycogen synthase kinase 3 (GSK3), a serine\\/threonine kinase that marks

Jesús Espada; Héctor Peinado; Manel Esteller; Amparo Cano

2005-01-01

129

A permissive function of phosphoinositide 3-kinase in Ras activation mediated by inhibition of GTPase-activating proteins  

Microsoft Academic Search

The activation status of the guanosine triphosphate (GTP)-binding protein Ras is dictated by the relative intensities of two opposing reactions: the formation of active Ras–GTP complexes, promoted by guanine-nucleotide exchange factors (GEFs), and their conversion to inactive Ras–GDP as a result of the deactivating action of GTPase-activating proteins (GAPs). The relevance of phosphoinositide 3-kinase (PI 3-kinase) to these processes is

Ignacio Rubio; Reinhard Wetzker

2000-01-01

130

Dibutyltin dilaurate induced thymic atrophy and modulation of phosphoinositide pathway of cell signalling in thymocytes of rats.  

PubMed

A marked dose dependent reduction in thymus weight and its nucleated cell counts with histological alterations was observed in rats exposed to oral dibutyltin dilaurate (DBTL) for 2 weeks at 2, 4, 8 or 16 mg/kg body weight. The incorporation of [3H]-inositol into all the three major phosphoinositides was drastically reduced in thymocytes in a dose dependent manner. Furthermore, the basal and the mitogen (Con A) stimulated [3H]-inositol phosphates generation was diminished significantly in 8 mg DBTL group. However, in vitro incubation of DBTL with thymocytes failed to evoke any change in phosphoinositide hydrolysis. Similarly, a time and dose dependent inhibition in phosphoinositide synthesis with as high as 80% by 10 microM DBTL was exhibited under in vitro conditions. A 130% and 600% enhancement of protein kinase C (PKC) activity in thymocytes was seen in 4 mg and 8 mg DBTL group, respectively. Addition of DBTL to the cell free assay system of thymocytes resulted in a concentration dependent activation of the enzyme activity. A dose dependent increase in intracellular calcium was also evident when DBTL was added to thymocytes under in vitro conditions. These results are of significance and may bear close relationship to the observed thymic atrophy by DBTL. PMID:7876466

Subramoniam, A; Khandelwal, S; Dwivedi, P D; Khanna, S; Shanker, R

1994-11-01

131

Galphaq binds to p110alpha/p85alpha phosphoinositide 3-kinase and displaces Ras.  

PubMed

Several studies have reported that activation of G(q)-coupled receptors inhibits PI3K (phosphoinositide 3-kinase) signalling. In the present study, we used purified proteins to demonstrate that Galpha(q) directly inhibits p110alpha/p85alpha PI3K in a GTP-dependent manner. Activated Galpha(q) binds to the p110alpha/p85alpha PI3K with an apparent affinity that is seven times stronger than that for Galpha(q).GDP as measured by fluorescence spectroscopy. In contrast, Galpha(q) did not bind to the p110gamma PI3K. Fluorescence spectroscopy experiments also showed that Galpha(q) competes with Ras, a PI3K activator, for binding to p110alpha/p85alpha. Interestingly, co-precipitation studies using deletion mutants showed that Galpha(q) binds to the p85-binding domain of p110alpha and not to the Ras-binding domain. Expression of constitutively active Galpha(q)Q209L in cells inhibited Ras activation of the PI3K/Akt pathway but had no effect on Ras/Raf/MEK [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase] signalling. These results suggest that activation of G(q)-coupled receptors leads to increased binding of Galpha(q).GTP to some isoforms of PI3K, which might explain why these receptors inhibit this signalling pathway in certain cell types. PMID:16268778

Ballou, Lisa M; Chattopadhyay, Mohar; Li, Yan; Scarlata, Suzanne; Lin, Richard Z

2006-03-15

132

Effects of Novel Isoform-Selective Phosphoinositide 3-Kinase Inhibitors on Natural Killer Cell Function  

PubMed Central

Phosphoinositide 3-kinases (PI3Ks) are promising targets for therapeutic development in cancer. The class I PI3K isoform p110? has received considerable attention in oncology because the gene encoding p110? (PIK3CA) is frequently mutated in human cancer. However, little is known about the function of p110? in lymphocyte populations that modulate tumorigenesis. We used recently developed investigational inhibitors to compare the function of p110? and other isoforms in natural killer (NK) cells, a key cell type for immunosurveillance and tumor immunotherapy. Inhibitors of all class I isoforms (pan-PI3K) significantly impaired NK cell-mediated cytotoxicity and antibody-dependent cellular cytotoxicity against tumor cells, whereas p110?-selective inhibitors had no effect. In NK cells stimulated through NKG2D, p110? inhibition modestly reduced PI3K signaling output as measured by AKT phosphorylation. Production of IFN-? and NK cell-derived chemokines was blocked by a pan-PI3K inhibitor and partially reduced by a p110?inhibitor, with lesser effects of p110? inhibitors. Oral administration of mice with MLN1117, a p110? inhibitor in oncology clinical trials, had negligible effects on NK subset maturation or terminal subset commitment. Collectively, these results support the targeting of PIK3CA mutant tumors with selective p110? inhibitors to preserve NK cell function. PMID:24915189

Yea, Sung Su; So, Lomon; Mallya, Sharmila; Lee, Jongdae; Rajasekaran, Kamalakannan; Malarkannan, Subramaniam; Fruman, David A.

2014-01-01

133

[Comparative-biochemical study of phosphoinositides as functionally active components of excitable nerve fiber membranes].  

PubMed

The effect of acetylcholine (ACh) and cyclic 3',5'-adenosine monophosphate (cAMP) on phosphoinositide (PI) metabolism and associated changes of the permeability of the nervous fibers in the crustaceans Carcinum maenas and Eriphia spinifrons, as well as in the frog Rana temporaria, has been investigated. It was shown that ACh induces a significant decrease in the content of triphosphoinositides (TPI) in the nervous fibers of the crabs and increases their potassium permeability, which in its turn results in depolarization of the fibers. ACh did not affect frog nervous fibers. cAMP significantly increases the incorporation of 32P into TPI fraction of crab nervous fibers during conduction of excitation and also facilitates repolarization of the fibers after application of ACh. Proserine effectively protects nervous fibers from the influence of ACh, indicating possible participation of acetylcholinesterase in TPI hydrolysis. The data obtained suggest that PI are involved into the regulation of membrane permeability of crab nervous fibers to potassium ions. PMID:194437

Tret'iak, A G; Limarenko, I M

1977-01-01

134

Unique serotonin receptor (5HT-1C) in choroid plexus is linked to phosphoinositide hydrolysis  

SciTech Connect

The binding of /sup 125/I-LSD to the 5HT-1C site and of /sup 3/H-ket-anserin to the 5HT-2 site was determined in choroid plexus and cerebral cortex of male Sprague-Dawley rats, respectively. As an index of phosphoinositide (PI) hydrolysis, whole choroid plexus and cerebral cortex slices were prelabelled with /sup 3/H-inositol and serotonin (5HT) stimulated release of /sup 3/H-inositol-1-phosphate was measured. 5HT stimulated PI hydrolysis in choroid plexus (6-fold) and in cerebral cortex (2.5-fold). 5HT was more potent in choroid plexus (EC/sub 50/ = 46 nM) consistent with the involvement of the 5HT-1C site. 5HT antagonists, ketanserin, mianserin and spiperone, inhibited the response to 5HT with different potencies in the two tissues. In cerebral cortex all 3 antagonists had nM affinities and a rank order (spiperone > ketanserin > mianserin) consistent with the 5HT-2 site. In choroid plexus, however, the rank order (mianserin > ketanserin > spiperone) and absolute potencies agreed with binding to the 5HT-1C site. These data suggest that the 5HT-1C site is a functional receptor which utilizes PI hydrolysis as its biochemical effector system.

Sanders-Bush, E.; Conn, P.J.; Hoffman, B.J.; Hartig, P.R.

1986-03-01

135

Nonenzymatic domains of Kalirin7 contribute to spine morphogenesis through interactions with phosphoinositides and Abl  

PubMed Central

Like several Rho GDP/GTP exchange factors (GEFs), Kalirin7 (Kal7) contains an N-terminal Sec14 domain and multiple spectrin repeats. A natural splice variant of Kalrn lacking the Sec14 domain and four spectrin repeats is unable to increase spine formation; our goal was to understand the function of the Sec14 and spectrin repeat domains. Kal7 lacking its Sec14 domain still increased spine formation, but the spines were short. Strikingly, Kal7 truncation mutants containing only the Sec14 domain and several spectrin repeats increased spine formation. The Sec14 domain bound phosphoinositides, a minor but crucial component of cellular membranes, and binding was increased by a phosphomimetic mutation. Expression of KalSec14-GFP in nonneuronal cells impaired receptor-mediated endocytosis, linking Kal7 to membrane trafficking. Consistent with genetic studies placing Abl, a non–receptor tyrosine kinase, and the Drosophila orthologue of Kalrn into the same signaling pathway, Abl1 phosphorylated two sites in the fourth spectrin repeat of Kalirin, increasing its sensitivity to calpain-mediated degradation. Treating cortical neurons of the wild-type mouse, but not the Kal7KO mouse, with an Abl inhibitor caused an increase in linear spine density. Phosphorylation of multiple sites in the N-terminal Sec14/spectrin region of Kal7 may allow coordination of the many signaling pathways contributing to spine morphogenesis. PMID:24600045

Ma, Xin-Ming; Miller, Megan B.; Vishwanatha, K. S.; Gross, Maegan J.; Wang, Yanping; Abbott, Thomas; Lam, TuKiet T.; Mains, Richard E.; Eipper, Betty A.

2014-01-01

136

Voltage sensitive phosphoinositide phosphatases of Xenopus: their tissue distribution and voltage dependence  

PubMed Central

Voltage sensitive phosphatases (VSPs) are unique proteins in which membrane potential controls enzyme activity. They are comprised of the voltage sensor domain of an ion channel coupled to a lipid phosphatase specific for phosphoinositides, and for ascidian and zebrafish VSPs, the phosphatase activity has been found to be activated by membrane depolarization. The physiological functions of these proteins are unknown, but their expression in testis and embryos suggests a role in fertilization or development. Here we investigate the expression pattern and voltage dependence of VSPs in two frog species, Xenopus laevis and Xenopus tropicalis, that are well suited for experimental studies of these possible functions. X. laevis has two VSP genes (Xl-VSP1 and Xl-VSP2), whereas X. tropicalis has only one gene (Xt-VSP). The highest expression of these genes was observed in testis, ovary, liver, and kidney. Our results show that while Xl-VSP2 activates only at positive membrane potentials outside of the physiological range, Xl-VSP1 and Xt-VSP phosphatase activity is regulated in the voltage range that regulates sperm-egg fusion at fertilization. PMID:21618529

Ratzan, William J.; Evsikov, Alexei V.; Okamura, Yasushi; Jaffe, Laurinda A.

2011-01-01

137

Evaluation of variation in the phosphoinositide-3-kinase catalytic subunit alpha oncogene and breast cancer risk  

PubMed Central

Background: Somatic mutations in phosphoinositide-3-kinase catalytic subunit alpha (PIK3CA) are frequent in breast tumours and have been associated with oestrogen receptor (ER) expression, human epidermal growth factor receptor-2 overexpression, lymph node metastasis and poor survival. The goal of this study was to evaluate the association between inherited variation in this oncogene and risk of breast cancer. Methods: A single-nucleotide polymorphism from the PIK3CA locus that was associated with breast cancer in a study of Caucasian breast cancer cases and controls from the Mayo Clinic (MCBCS) was genotyped in 5436 cases and 5280 controls from the Cancer Genetic Markers of Susceptibility (CGEMS) study and in 30?949 cases and 29?788 controls from the Breast Cancer Association Consortium (BCAC). Results: Rs1607237 was significantly associated with a decreased risk of breast cancer in MCBCS, CGEMS and all studies of white Europeans combined (odds ratio (OR)=0.97, 95% confidence interval (CI) 0.95–0.99, P=4.6 × 10?3), but did not reach significance in the BCAC replication study alone (OR=0.98, 95% CI 0.96–1.01, P=0.139). Conclusion: Common germline variation in PIK3CA does not have a strong influence on the risk of breast cancer PMID:22033276

Stevens, K N; Garcia-Closas, M; Fredericksen, Z; Kosel, M; Pankratz, V S; Hopper, J L; Dite, G S; Apicella, C; Southey, M C; Schmidt, M K; Broeks, A; Van 't Veer, L J; Tollenaar, R A E M; Fasching, P A; Beckmann, M W; Hein, A; Ekici, A B; Johnson, N; Peto, J; dos Santos Silva, I; Gibson, L; Sawyer, E; Tomlinson, I; Kerin, M J; Chanock, S; Lissowska, J; Hunter, D J; Hoover, R N; Thomas, G D; Milne, R L; Perez, JI Arias; Gonzalez-Neira, A; Benitez, J; Burwinkel, B; Meindl, A; Schmutzler, R K; Bartrar, C R; Hamann, U; Ko, Y D; Bruning, T; Chang-Claude, J; Hein, R; Wang-Gohrke, S; Dork, T; Schurmann, P; Bremer, M; Hillemanns, P; Bogdanova, N; Zalutsky, J V; Rogov, Y I; Antonenkova, N; Lindblom, A; Margolin, S; Mannermaa, A; Kataja, V; Kosma, V-M; Hartikainen, J; Chenevix-Trench, G; Chen, X; Peterlongo, P; Bonanni, B; Bernard, L; Manoukian, S; Wang, X; Cerhan, J; Vachon, C M; Olson, J; Giles, G G; Baglietto, L; McLean, C A; Severi, G; John, E M; Miron, A; Winqvist, R; Pylkas, K; Jukkola-Vuorinen, A; Grip, M; Andrulis, I; Knight, J A; Glendon, G; Mulligan, A M; Cox, A; Brock, I W; Elliott, G; Cross, S S; Pharoah, P P; Dunning, A M; Pooley, K A; Humphreys, M K; Wang, J; Kang, D; Yoo, K-Y; Noh, D-Y; Sangrajrang, S; Gabrieau, V; Brennan, P; McKay, J; Anton-Culver, H; Ziogas, A; Couch, F J; Easton, D F

2011-01-01

138

Calcium mobilization and phosphoinositide turnover in fluoride-activated human neutrophils  

SciTech Connect

Fluoride ion, at concentrations above 10 mM, has been found to activate a superoxide production response in human neutrophils which is strongly dependent on the presence of extracellular calcium. In an attempt to further explore the calcium requirement of fluoride-induced neutrophil activation, intracellular calcium concentrations were monitored through use of the fluorescent calcium probe, Quin 2. Fluoride ion, at concentrations between 10 and 20 mM, was found to elicit a rise in intracellular calcium levels which was characterized by a lag period of 4 to 10 min and a prolonged duration of action (greater than 20 min). In contrast, the chemotactic peptide, formylmethionyl-leucyl-phenylalanine (FMLP), induced a rise in intracellular calcium concentration which peaked within 1 min. Preincubation of cells with 1 ..mu..g/ml pertussis toxin resulted in inhibition of the FMLP-induced response, but not that elicited by fluoride. Furthermore, anion exchange chromatography indicated that inositol phosphate accumulation occurred in fluoride-treated cells in association with calcium mobilization. Recent evidence suggests that the FMLP receptor is coupled to phospholipase C and phosphoinositide turnover through a guanine nucleotide binding protein susceptible to inhibition by pertussis toxin. Present results suggest that fluoride ion may serve to activate this protein in a manner resistant to inhibition by pertussis toxin.

Strnad, C.F.; Wong, K.

1986-05-01

139

Molecular cloning and biochemical characterization of a Drosophila phosphatidylinositol-specific phosphoinositide 3-kinase.  

PubMed Central

Molecular, biochemical and genetic characterization of phosphoinositide 3-kinases (PI3Ks) have identified distinct classes of enzymes involved in processes mediated by activation of cell-surface receptors and in constitutive intracellular protein trafficking events. The latter process appears to involve a PtdIns-specific PI3K first described in yeast as a mutant, vps34, defective in the sorting of newly synthesized proteins from the Golgi to the vacuole. We have identified a representative member of each class of PI3Ks in Drosophila using a PCR-based approach. In the present paper we describe the molecular cloning of a PI3K from Drosophila, P13K_59F, that shows sequence similarity to Vps34. PI3K_59F encodes a protein of 108 kDa co-linear with Vps34 homologues, and with three regions of sequence similarity to other PI3Ks. Biochemical characterization of the enzyme, by expression of the complete coding sequence as a glutathione S-transferase fusion protein in Sf9 cells, demonstrates that PI3K_59F is a PtdIns-specific PI3K that can utilize either Mg2+ or Mn2+. This activity is sensitive to inhibition both by non-ionic detergent (Nonidet P40) and by wortmannin (IC50 10 nM). PI3K_59F, therefore, conserves both the structural and biochemical properties of the Vps34 class of enzymes. PMID:9032475

Linassier, C; MacDougall, L K; Domin, J; Waterfield, M D

1997-01-01

140

Phosphoinositide-3 kinase ? required for LPS-induced transepithelial neutrophil trafficking in the lung  

PubMed Central

Phosphoinositide 3-kinase ? (PI3K?) is a critical mediator of directional cell movement. Here, we sought to characterize the role of PI3K? in mediating the different steps of PMN trafficking in the lung. In a murine model of LPS-induced lung injury, PMN migration into the different lung compartments was determined in PI3K? gene-deficient (PI3K??/?) and wildtype mice. Bone marrow chimeras were created to characterize the role of PI3K? on hematopoietic vs. non-hematopoietic cells. A small molecule PI3K? inhibitor was tested in vitro and in vivo. PMN adhesion to the pulmonary endothelium and transendothelial migration into the lung interstitium was enhanced in PI3K??/? mice. However, transepithelial migration into the alveolar space was reduced in these mice. When irradiated PI3K??/? mice were reconstituted with bone marrow from wildtype mice, migratory activity into the alveolar space was restored partially. A small molecule PI3K? inhibitor reduced chemokine-induced PMN migration in vitro when PMNs or epithelial cells but not when endothelial cells were treated. The inhibitor also reduced LPS-induced PMN migration in vivo. We conclude that PI3K? is required for transepithelial but not for transendothelial migration in LPS-induced lung injury. Inhibition of PI3K? activity may be effective at curbing excessive PMN infiltration in lung injury. PMID:19797129

Reutershan, Jorg; Saprito, Mary S.; Wu, Dan; Ruckle, Thomas; Ley, Klaus

2009-01-01

141

Detection of myotubularin phosphatases activity on phosphoinositides in vitro and ex vivo.  

PubMed

Phosphoinositides (PPIn) are important regulators of cellular processes like intracellular protein transport, cellular proliferation, apoptosis, and cytoskeletal organization. The amount and localization of these membrane-bound second messengers are regulated through a set of specific phospholipases, lipid kinases, and phosphatases. The elucidation of PPIn-phosphatases and their cellular function has gained much attention because phosphatase dysregulation is often associated with human genetic diseases. Our laboratory has identified the 3'-PPIn-phosphatase myotubularin 1 (MTM1) mutated in X-linked myotubular myopathy (XLMTM). In addition, a whole family of myotubularin-related proteins (MTMR1-MTMR13) has been discovered. Some of them display phosphatase activity, whereas for other family members no enzymatic activity could be detected. Nevertheless, these "dead phosphatases" myotubularins are conserved throughout evolution and probably exert regulatory function by heteromeric interaction with active phosphatase members. It was shown that MTM1 and related phosphatases act on PtdIns3P and PtdIns(3,5)P2; both PPIn species are important regulators of endocytic pathways. We describe two methods to determine phosphatase activity and substrate specificity of myotubularins. One is an immunoprecipitation-phosphatase assay, testing the activity of myotubularin immunoprecipitated from overexpressing cells on artificial PPIn. The other method analyzes phosphatase activity indirectly ex vivo in transiently transfected mammalian cells. The presence and subcellular localization of the myotubularin substrate PtdIns3P were determined using a specific binding domain (2xFYVE) produced recombinantly as a biosensor. PMID:19160676

Rohde, Holger Maria; Tronchère, Hélène; Payrastre, Bernard; Laporte, Jocelyn

2009-01-01

142

The Phosphoinositide-3-Kinase-Akt-mTOR Pathway as a Therapeutic Target in Breast Cancer  

PubMed Central

The phosphoinositide-3-kinase (PI3-kinase)-Akt-mTOR pathway is a central signal transduction pathway that regulates many critical aspects of normal and cancer physiology, including cell proliferation, apoptosis, cell morphology and migration, protein synthesis, and integration of metabolism. In breast cancer, somatic mutations that activate the pathway occur in more than 50% of tumors, underscoring the potentially broad impact of targeting the pathway for therapy. A vast body of preclinical data demonstrates the efficacy of pathway inhibition on tumor growth, and evidence also shows that activation of the pathway occurs in models of acquired resistance to hormonal therapy. This preclinical work led to the investigation of allosteric mTOR inhibitors, everolimus and temsirolimus, in metastatic hormone receptor–positive breast cancer. The recent BOLERO-2 trial comparing everolimus plus exemestane versus placebo plus exemestane in women with resistance to nonsteroidal aromatase inhibitors demonstrated a 6-month improvement in progression-free survival and led to FDA approval of everolimus for this indication in the United States. This landmark trial is the first demonstration of significant clinical benefit using drugs targeting this pathway in breast cancer. Many questions remain about the role of everolimus and other pathway-targeting drugs in clinical development in breast cancer treatment. This article reviews the role of the PI3-kinase-Akt-mTOR pathway in breast cancer biology and the clinical trial evidence available to date. PMID:23744866

Lauring, Josh; Park, Ben Ho; Wolff, Antonio C.

2014-01-01

143

Nuclear but Not Cytosolic Phosphoinositide 3-Kinase Beta Has an Essential Function in Cell Survival ?  

PubMed Central

Class IA phosphoinositide 3-kinases (PI3Ks) are heterodimeric enzymes composed of a p85 regulatory and a p110 catalytic subunit that induce the formation of 3-polyphosphoinositides, which mediate cell survival, division, and migration. There are two ubiquitous PI3K isoforms p110? and p110? that have nonredundant functions in embryonic development and cell division. However, whereas p110? concentrates in the cytoplasm, p110? localizes to the nucleus and modulates nuclear processes such as DNA replication and repair. At present, the structural features that determine p110? nuclear localization remain unknown. We describe here that association with the p85? regulatory subunit controls p110? nuclear localization. We identified a nuclear localization signal (NLS) in p110? C2 domain that mediates its nuclear entry, as well as a nuclear export sequence (NES) in p85?. Deletion of p110? induced apoptosis, and complementation with the cytoplasmic C2-NLS p110? mutant was unable to restore cell survival. These studies show that p110? NLS and p85? NES regulate p85?/p110? nuclear localization, supporting the idea that nuclear, but not cytoplasmic, p110? controls cell survival. PMID:21383062

Kumar, Amit; Redondo-Muñoz, Javier; Perez-García, Vicente; Cortes, Isabel; Chagoyen, Monica; Carrera, Ana C.

2011-01-01

144

Class IA Phosphoinositide 3-Kinase Regulates Heart Size and Physiological Cardiac Hypertrophy  

PubMed Central

Class IA phosphoinositide 3-kinases (PI3Ks) are activated by growth factor receptors, and they regulate, among other processes, cell growth and organ size. Studies using transgenic mice overexpressing constitutively active and dominant negative forms of the p110? catalytic subunit of class IA PI3K have implicated the role of this enzyme in regulating heart size and physiological cardiac hypertrophy. To further understand the role?of class IA PI3K in controlling heart growth and to circumvent potential complications from the overexpression of dominant negative and constitutively active proteins, we generated mice with muscle-specific deletion of the p85? regulatory subunit and germ line deletion of the p85? regulatory subunit of class IA PI3K. Here we show that mice with cardiac deletion of both p85 subunits exhibit attenuated Akt signaling in the heart, reduced heart size, and altered cardiac gene expression. Furthermore, exercise-induced cardiac hypertrophy is also attenuated in the p85 knockout hearts. Despite such defects in postnatal developmental growth and physiological hypertrophy, the p85 knockout hearts exhibit normal contractility and myocardial histology. Our results therefore provide strong genetic evidence that class IA PI3Ks are critical regulators for the developmental growth and physiological hypertrophy of the heart. PMID:16227599

Luo, Ji; McMullen, Julie R.; Sobkiw, Cassandra L.; Zhang, Li; Dorfman, Adam L.; Sherwood, Megan C.; Logsdon, M. Nicole; Horner, James W.; DePinho, Ronald A.; Izumo, Seigo; Cantley, Lewis C.

2005-01-01

145

Critical role for phosphoinositide 3-kinase gamma in parasite invasion and disease progression of cutaneous leishmaniasis.  

PubMed

Obligate intracellular pathogens such as Leishmania specifically target host phagocytes for survival and replication. Phosphoinositide 3-kinase ? (PI3K?), a member of the class I PI3Ks that is highly expressed by leukocytes, controls cell migration by initiating actin polymerization and cytoskeletal reorganization, which are processes also critical for phagocytosis. In this study, we demonstrate that class IB PI3K, PI3K?, plays a critical role in pathogenesis of chronic cutaneous leishmaniasis caused by L. mexicana. Using the isoform-selective PI3K? inhibitor, AS-605240 and PI3K? gene-deficient mice, we show that selective blockade or deficiency of PI3K? significantly enhances resistance against L. mexicana that is associated with a significant suppression of parasite entry into phagocytes and reduction in recruitment of host phagocytes as well as regulatory T cells to the site of infection. Furthermore, we demonstrate that AS-605240 is as effective as the standard antileishmanial drug sodium stibogluconate in treatment of cutaneous leishmaniasis caused by L. mexicana. These findings reveal a unique role for PI3K? in Leishmania invasion and establishment of chronic infection, and demonstrate that therapeutic targeting of host pathways involved in establishment of infection may be a viable strategy for treating infections caused by obligate intracellular pathogens such as Leishmania. PMID:22232690

Cummings, Hannah E; Barbi, Joseph; Reville, Patrick; Oghumu, Steve; Zorko, Nicholas; Sarkar, Anasuya; Keiser, Tracy L; Lu, Bao; Rückle, Thomas; Varikuti, Sanjay; Lezama-Davila, Claudio; Wewers, Mark D; Whitacre, Caroline; Radzioch, Danuta; Rommel, Christian; Seveau, Stéphanie; Satoskar, Abhay R

2012-01-24

146

Phosphoinositide 3-kinase ? gene mutation predisposes to respiratory infection and airway damage  

PubMed Central

Genetic mutations cause primary immunodeficiencies (PIDs), which predispose to infections. Here we describe Activated PI3K-? Syndrome (APDS), a PID associated with a dominant gain-of-function mutation E1021K in the p110? protein, the catalytic subunit of phosphoinositide 3-kinase ? (PI3K?), encoded by the PIK3CD gene. We found E1021K in 17 patients from seven unrelated families, but not among 3,346 healthy subjects. APDS was characterized by recurrent respiratory infections, progressive airway damage, lymphopenia, increased circulating transitional B cells, increased IgM and reduced IgG2 levels in serum and impaired vaccine responses. The E1021K mutation enhanced membrane association and kinase activity of p110?. Patient-derived lymphocytes had increased levels of phosphatidylinositol 3,4,5-trisphosphate and phosphorylated AKT protein and were prone to activation-induced cell death. Selective p110? inhibitors IC87114 and GS-1101 reduced the activity of the mutant enzyme in vitro, suggesting a therapeutic approach for patients with APDS. PMID:24136356

Angulo, Ivan; Vadas, Oscar; Garcon, Fabien; Banham-Hall, Edward; Plagnol, Vincent; Leahy, Timothy R.; Baxendale, Helen; Coulter, Tanya; Curtis, James; Wu, Changxin; Blake-Palmer, Katherine; Perisic, Olga; Smyth, Deborah; Maes, Mailis; Fiddler, Christine; Juss, Jatinder; Cilliers, Deirdre; Markelj, Gasper; Chandra, Anita; Farmer, George; Kielkowska, Anna; Clark, Jonathan; Kracker, Sven; Debre, Marianne; Picard, Capucine; Pellier, Isabelle; Jabado, Nada; Morris, James A.; Barcenas-Morales, Gabriela; Fischer, Alain; Stephens, Len; Hawkins, Phillip; Barrett, Jeffrey C.; Abinun, Mario; Clatworthy, Menna; Durandy, Anne; Doffinger, Rainer; Chilvers, Edwin; Cant, Andrew J.; Kumararatne, Dinakantha; Okkenhaug, Klaus; Williams, Roger L.; Condliffe, Alison; Nejentsev, Sergey

2014-01-01

147

Class I and class III phosphoinositide 3-kinases are required for actin polymerization that propels phagosomes.  

PubMed

Actin polymerization drives the extension of pseudopods that trap and engulf phagocytic targets. The polymerized actin subsequently dissociates as the phagocytic vacuole seals and detaches from the plasma membrane. We found that phagosomes formed by engagement of integrins that serve as complement receptors (CR3) undergo secondary waves of actin polymerization, leading to the formation of "comet tails" that propel the vacuoles inside the cells. Actin tail formation was accompanied by and required de novo formation of PI(3,4)P(2) and PI(3,4,5)P(3) on the phagosomal membrane by class I phosphoinositide 3-kinases (PI3Ks). Although the phosphatidylinositide phosphatase Inpp5B was recruited to nascent phagosomes, it rapidly detached from the membrane after phagosomes sealed. Detachment of Inpp5B required the formation of PI(3)P. Thus, class III PI3K activity was also required for the accumulation of PI(4,5)P(2) and PI(3,4,5)P(3) and for actin tail formation. These experiments reveal a new PI(3)P-sensitive pathway leading to PI(3,4)P(2) and PI(3,4,5)P(3) formation and signaling in endomembranes. PMID:21115805

Bohdanowicz, Michal; Cosío, Gabriela; Backer, Jonathan M; Grinstein, Sergio

2010-11-29

148

Class I and class III phosphoinositide 3-kinases are required for actin polymerization that propels phagosomes  

PubMed Central

Actin polymerization drives the extension of pseudopods that trap and engulf phagocytic targets. The polymerized actin subsequently dissociates as the phagocytic vacuole seals and detaches from the plasma membrane. We found that phagosomes formed by engagement of integrins that serve as complement receptors (CR3) undergo secondary waves of actin polymerization, leading to the formation of “comet tails” that propel the vacuoles inside the cells. Actin tail formation was accompanied by and required de novo formation of PI(3,4)P2 and PI(3,4,5)P3 on the phagosomal membrane by class I phosphoinositide 3-kinases (PI3Ks). Although the phosphatidylinositide phosphatase Inpp5B was recruited to nascent phagosomes, it rapidly detached from the membrane after phagosomes sealed. Detachment of Inpp5B required the formation of PI(3)P. Thus, class III PI3K activity was also required for the accumulation of PI(4,5)P2 and PI(3,4,5)P3 and for actin tail formation. These experiments reveal a new PI(3)P-sensitive pathway leading to PI(3,4)P2 and PI(3,4,5)P3 formation and signaling in endomembranes. PMID:21115805

Bohdanowicz, Michal; Cosío, Gabriela; Backer, Jonathan M.

2010-01-01

149

Identification of myo-inositol 1,2-cyclic monophosphate by electrospray tandem mass spectrometry, a major constituent of EGF-stimulated phosphoinositide turnover in MDA 468 cells.  

PubMed

Epidermal growth factor (EGF) caused an increase in phosphoinositide (PI) turnover in MDA 468 cells. This EGF-stimulated effect was inhibited by the protein tyrosine kinase inhibitor lavendustin A (LA). MDA 468 cells generated an atypical PI turnover profile. Examination and quantitation of the PI metabolite profile showed that even control cells produced a metabolite which was acid-labile and which formed about 60% of the total PI metabolites. By using the technique of electrospray ionization tandem mass spectrometry, we were able to confirm the identity of this acid-labile metabolite through the specific fragmentation as compared with the standard. The precursor molecule fragmented into two distinct productions with molar masses identical to that of the standard myo-inositol 1,2-cyclic monophosphate (cInsP). Changes in the PI turnover profile could be accounted for by the alterations in myo-inositol 1,2-cyclic monophosphate generated in these cells. We thus conclude that, by some as-yet-unidentified mechanism, cyclic inositol monophosphate forms a major constituent of EGF-stimulated PI turnover in MDA 468 cells. PMID:12083795

Mandal, Soma; Richardson, Vernon J; Banoub, Joseph; Church, Jon G

2002-07-15

150

Evolution of the Voltage Sensor Domain of the Voltage-Sensitive Phosphoinositide Phosphatase VSP/TPTE Suggests a Role as a Proton Channel in Eutherian Mammals  

PubMed Central

The voltage-sensitive phosphoinositide phosphatases provide a mechanism to couple changes in the transmembrane electrical potential to intracellular signal transduction pathways. These proteins share a domain architecture that is conserved in deuterostomes. However, gene duplication events in primates, including humans, give rise to the paralogs TPTE and TPTE2 that retain protein domain organization but, in the case of TPTE, have lost catalytic activity. Here, we present evidence that these human proteins contain a functional voltage sensor, similar to that in nonmammalian orthologs. However, domains of these human proteins can also generate a noninactivating outward current that is not observed in zebra fish or tunicate orthologs. This outward current has the anticipated characteristics of a voltage-sensitive proton current and is due to the appearance of a single histidine residue in the S4 transmembrane segment of the voltage sensor. Histidine is observed at this position only during the eutherian radiation. Domains from both human paralogs generate proton currents. This apparent gain of proton channel function during the evolution of the TPTE protein family may account for the conservation of voltage sensor domains despite the loss of phosphatase activity in some human paralogs. PMID:22396523

Sutton, Keith A.; Jungnickel, Melissa K.; Jovine, Luca; Florman, Harvey M.

2012-01-01

151

Protein Kinase Activity of Phosphoinositide 3-Kinase Regulates Cytokine-Dependent Cell Survival  

PubMed Central

The dual specificity protein/lipid kinase, phosphoinositide 3-kinase (PI3K), promotes growth factor-mediated cell survival and is frequently deregulated in cancer. However, in contrast to canonical lipid-kinase functions, the role of PI3K protein kinase activity in regulating cell survival is unknown. We have employed a novel approach to purify and pharmacologically profile protein kinases from primary human acute myeloid leukemia (AML) cells that phosphorylate serine residues in the cytoplasmic portion of cytokine receptors to promote hemopoietic cell survival. We have isolated a kinase activity that is able to directly phosphorylate Ser585 in the cytoplasmic domain of the interleukin 3 (IL-3) and granulocyte macrophage colony stimulating factor (GM-CSF) receptors and shown it to be PI3K. Physiological concentrations of cytokine in the picomolar range were sufficient for activating the protein kinase activity of PI3K leading to Ser585 phosphorylation and hemopoietic cell survival but did not activate PI3K lipid kinase signaling or promote proliferation. Blockade of PI3K lipid signaling by expression of the pleckstrin homology of Akt1 had no significant impact on the ability of picomolar concentrations of cytokine to promote hemopoietic cell survival. Furthermore, inducible expression of a mutant form of PI3K that is defective in lipid kinase activity but retains protein kinase activity was able to promote Ser585 phosphorylation and hemopoietic cell survival in the absence of cytokine. Blockade of p110? by RNA interference or multiple independent PI3K inhibitors not only blocked Ser585 phosphorylation in cytokine-dependent cells and primary human AML blasts, but also resulted in a block in survival signaling and cell death. Our findings demonstrate a new role for the protein kinase activity of PI3K in phosphorylating the cytoplasmic tail of the GM-CSF and IL-3 receptors to selectively regulate cell survival highlighting the importance of targeting such pathways in cancer. PMID:23526884

Green, Benjamin D.; Barry, Emma F.; Ma, Yuefang; Woodcock, Joanna; Fitter, Stephen; Zannettino, Andrew C. W.; Pitson, Stuart M.; Hughes, Timothy P.; Lopez, Angel F.; Shepherd, Peter R.; Wei, Andrew H.; Ekert, Paul G.; Guthridge, Mark A.

2013-01-01

152

Live-Cell Imaging of Phosphoinositide Dynamics and Membrane Architecture during Legionella Infection  

PubMed Central

ABSTRACT The causative agent of Legionnaires’ disease, Legionella pneumophila, replicates in amoebae and macrophages in a distinct membrane-bound compartment, the Legionella-containing vacuole (LCV). LCV formation is governed by the bacterial Icm/Dot type IV secretion system that translocates ~300 different “effector” proteins into host cells. Some of the translocated effectors anchor to the LCV membrane via phosphoinositide (PI) lipids. Here, we use the soil amoeba Dictyostelium discoideum, producing fluorescent PI probes, to analyze the LCV PI dynamics by live-cell imaging. Upon uptake of wild-type or Icm/Dot-deficient L. pneumophila, PtdIns(3,4,5)P3 transiently accumulated for an average of 40 s on early phagosomes, which acquired PtdIns(3)P within 1 min after uptake. Whereas phagosomes containing ?icmT mutant bacteria remained decorated with PtdIns(3)P, more than 80% of wild-type LCVs gradually lost this PI within 2 h. The process was accompanied by a major rearrangement of PtdIns(3)P-positive membranes condensing to the cell center. PtdIns(4)P transiently localized to early phagosomes harboring wild-type or ?icmT L. pneumophila and was cleared within minutes after uptake. During the following 2 h, PtdIns(4)P steadily accumulated only on wild-type LCVs, which maintained a discrete PtdIns(4)P identity spatially separated from calnexin-positive endoplasmic reticulum (ER) for at least 8 h. The separation of PtdIns(4)P-positive and ER membranes was even more pronounced for LCVs harboring ?sidC-sdcA mutant bacteria defective for ER recruitment, without affecting initial bacterial replication in the pathogen vacuole. These findings elucidate the temporal and spatial dynamics of PI lipids implicated in LCV formation and provide insight into host cell membrane and effector protein interactions. PMID:24473127

Weber, Stephen; Wagner, Maria; Hilbi, Hubert

2014-01-01

153

Changes in phosphoinositide turnover, Ca sup 2+ mobilization, and protein phosphorylation in platelets from NIDDM patients  

SciTech Connect

Enhanced platelet functions have been demonstrated in patients with non-insulin-dependent diabetes mellitus (NIDDM). This study evaluated abnormalities in platelet signal transduction in diabetic patients, including turnover of phosphoinositides, mobilization of intracellular Ca2+, and phosphorylation of 20,000- and 47,000-Mr proteins (P20 and P47). Washed platelets were obtained from 6 patients with NIDDM whose platelet aggregation rates were abnormally elevated (DM-A group), 11 NIDDM patients with normal platelet aggregation rates (DM-B group), and 8 age-matched healthy control subjects. The mass and specific radioactivity of phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol (PI), and phosphatidic acid (PA) in 32P-labeled platelets were not different among the three groups. Hydrolysis of PIP2, PIP, and PI; accumulation of PA; and phosphorylation of P20 in platelets stimulated by 0.05 U/ml thrombin were significantly increased in the DM-A group compared with the control or DM-B group. There was no difference in P47 phosphorylation among the three groups. On the contrary, P20 and P47 phosphorylation induced by 50 nM of 12-O-tetradecanoylphorbol-13-acetate, an activator of protein kinase C, was significantly decreased in the DM-A group. Additionally, the intracellular free Ca2+ concentration (( Ca2+)i) was measured with the fluorescent Ca2+ indicator fura 2. Although the basal (Ca2+)i value was similar in the three groups, the rise in (Ca2+)i induced by 0.05 U/ml thrombin in the presence and the absence of extracellular Ca2+ was significantly higher in the DM-A group than the other groups.

Ishii, H.; Umeda, F.; Hashimoto, T.; Nawata, H. (Kyushu Univ., Fukuoka (Japan))

1990-12-01

154

Phosphoinositide-3 Kinase-Akt Pathway Controls Cellular Entry of Ebola Virus  

E-print Network

The phosphoinositide-3 kinase (PI3K) pathway regulates diverse cellular activities related to cell growth, migration, survival, and vesicular trafficking. It is known that Ebola virus requires endocytosis to establish an infection. However, the cellular signals that mediate this uptake were unknown for Ebola virus as well as many other viruses. Here, the involvement of PI3K in Ebola virus entry was studied. A novel and critical role of the PI3K signaling pathway was demonstrated in cell entry of Zaire Ebola virus (ZEBOV). Inhibitors of PI3K and Akt significantly reduced infection by ZEBOV at an early step during the replication cycle. Furthermore, phosphorylation of Akt-1 was induced shortly after exposure of cells to radiation-inactivated ZEBOV, indicating that the virus actively induces the PI3K pathway and that replication was not required for this induction. Subsequent use of pseudotyped Ebola virus and/or Ebola virus-like particles, in a novel virus entry assay, provided evidence that activity of PI3K/Akt is required at the virus entry step. Class 1A PI3Ks appear to play a predominant role in regulating ZEBOV entry, and Rac1 is a key downstream effector in this regulatory cascade. Confocal imaging of fluorescently labeled ZEBOV indicated that inhibition of PI3K, Akt, or Rac1 disrupted normal uptake of virus particles into cells and resulted in aberrant accumulation of virus into a cytosolic compartment that was non-permissive for membrane fusion. We conclude that PI3K-mediated signaling plays an important role in regulating vesicular trafficking of ZEBOV necessary for cell entry. Disruption of this signaling leads to inappropriate trafficking within the cell and a block in steps leading to membrane

Mohammad F. Saeed; Andrey A. Kolokoltsov; Er N. Freiberg; Michael R. Holbrook; Robert A. Davey

155

Role of phosphoinositide 3-kinase in the pathogenesis of acute pancreatitis  

PubMed Central

A large body of experimental and clinical data supports the notion that inflammation in acute pancreatitis has a crucial role in the pathogenesis of local and systemic damage and is a major determinant of clinical severity. Thus, research has recently focused on molecules that can regulate the inflammatory processes, such as phosphoinositide 3-kinases (PI3Ks), a family of lipid and protein kinases involved in intracellular signal transduction. Studies using genetic ablation or pharmacologic inhibitors of different PI3K isoforms, in particular the class I PI3K? and PI3K?, have contributed to a greater understanding of the roles of these kinases in the modulation of inflammatory and immune responses. Recent data suggest that PI3Ks are also involved in the pathogenesis of acute pancreatitis. Activation of the PI3K signaling pathway, and in particular of the class IB PI3K? isoform, has a significant role in those events which are necessary for the initiation of acute pancreatic injury, namely calcium signaling alteration, trypsinogen activation, and nuclear factor-?B transcription. Moreover, PI3K? is instrumental in modulating acinar cell apoptosis, and regulating local neutrophil infiltration and systemic inflammatory responses during the course of experimental acute pancreatitis. The availability of PI3K inhibitors selective for specific isoforms may provide new valuable therapeutic strategies to improve the clinical course of this disease. This article presents a brief summary of PI3K structure and function, and highlights recent advances that implicate PI3Ks in the pathogenesis of acute pancreatitis. PMID:25386068

Lupia, Enrico; Pigozzi, Luca; Goffi, Alberto; Hirsch, Emilio; Montrucchio, Giuseppe

2014-01-01

156

Phosphoinositides Play Differential Roles in Regulating Phototropin1- and Phototropin2-Mediated Chloroplast Movements in Arabidopsis  

PubMed Central

Phototropins are UVA/blue-light receptors involved in controlling the light-dependent physiological responses which serve to optimize the photosynthetic activity of plants and promote growth. The phototropin-induced phosphoinositide (PI) metabolism has been shown to be essential for stomatal opening and phototropism. However, the role of PIs in phototropin-induced chloroplast movements remains poorly understood. The aim of this work is to determine which PI species are involved in the control of chloroplast movements in Arabidopsis and the nature of their involvement. We present the effects of the inactivation of phospholipase C (PLC), PI3-kinase (PI3K) and PI4-kinase (PI4K) on chloroplast relocations in Arabidopsis. The inhibition of the phosphatidylinositol 4,5-bisphospahte [PI(4,5)P2]-PLC pathway, using neomycin and U73122, suppressed the phot2-mediated chloroplast accumulation and avoidance responses, without affecting movement responses controlled by phot1. On the other hand, PI3K and PI4K activities are more restricted to phot1- and phot2-induced weak-light responses. The inactivation of PI3K and PI4K by wortmannin and LY294002 severely affected the weak blue-light-activated accumulation response but had little effect on the strong blue-light-activated avoidance response. The inhibitory effect observed with PI metabolism inhibitors is, at least partly, due to a disturbance in Ca2+(c) signaling. Using the transgenic aequorin system, we show that the application of these inhibitors suppresses the blue-light-induced transient Ca2+(c) rise. These results demonstrate the importance of PIs in chloroplast movements, with the PI(4,5)P2-PLC pathway involved in phot2 signaling while PI3K and PI4K are required for the phot1- and phot2-induced accumulation response. Our results suggest that these PIs modulate cytosolic Ca2+ signaling during movements. PMID:23405144

Aggarwal, Chhavi; Labuz, Justyna; Gabrys, Halina

2013-01-01

157

Peroxovanadate induces tyrosine phosphorylation of phosphoinositide-dependent protein kinase-1 potential involvement of src kinase.  

PubMed

Phosphoinositide-dependent protein kinase-1 (PDK1) is a recently identified kinase that phosphorylates and activates protein kinase B (PKB). Activation of PKB by insulin is linked to its translocation from the cytosol to the plasma membrane. However, no data are available yet concerning the localization of PDK1 in insulin-sensitive tissue. Using isolated adipocytes, we studied the effect of insulin and of an insulin-mimicking agent peroxovanadate on the subcellular localization of PDK1. In unstimulated adipocytes, overexpressed PDK1 was mostly cytosolic with a low amount associated to membranes. Peroxovanadate stimulation induced the redistribution of PDK1 to the membranes while insulin was without effect. This peroxovanadate effect was dependent on phosphatidylinositol 3,4,5 triphosphate [PtdIns(3,4,5)P3] production as inhibition of PtdIns 3-kinase by wortmannin or deletion of the PH domain of PDK1 prevented the peroxovanadate-induced translocation of PDK1. Further, peroxovanadate-treatment induced a tyrosine phosphorylation of PDK1 which was wortmannin insensitive and did not require the PH domain of PDK1. An inhibitor of Src kinase (PP2) decreased the peroxovanadate-induced PDK1 tyrosine phosphorylation and overexpression of v-Src stimulated this phosphorylation. Mutation of tyrosine 373 of PDK1 abolished the v-Src induced PDK1 tyrosine phosphorylation and partially reduced the effect of peroxovanadate. Our findings suggest that PDK1 could be a substrate for tyrosine kinases and identify Src kinase as one of the tyrosine kinases able to phosphorylate PDK1. PMID:11054117

Grillo, S; Grémeaux, T; Casamayor, A; Alessi, D R; Le Marchand-Brustel, Y; Tanti, J F

2000-11-01

158

The phosphoinositide-dependent protein kinase 1 inhibitor, UCN-01, induces fragmentation: Possible role of metalloproteinases.  

PubMed

Phosphoinositide-dependent protein kinase 1 (PDK1) is a key enzyme, master regulator of cellular proliferation and metabolism; it is considered a key target for pharmacological intervention. Using membranes obtained from DDT1 MF-2 cells, phospho-PDK1 was identified by Western blotting, as two major protein bands of Mr 58-68kDa. Cell incubation with the PDK1 inhibitor, UCN-01, induced a time- and concentration-dependent decrease in the amount of phospho-PDK1 with a concomitant appearance of a ?42kDa phosphorylated fragment. Knocking down PDK1 diminished the amount of phospho-PDK1 detected in membranes, accompanied by similarly decreased fragment generation. UCN-01-induced fragment generation was also observed in membranes from cells stably expressing a myc-tagged PDK1 construct. Other PDK1 inhibitors were also tested: OSU-03012 induced a clear decrease in phospho-PDK1 and increased the presence of the phosphorylated fragment in membrane preparations; in contrast, GSK2334470 and staurosporine induced only marginal increases in the amount of PDK1 fragment. Galardin and batimastat, two metalloproteinase inhibitors, markedly attenuated inhibitor-induced PDK1 fragment generation. Metalloproteinases 2, 3, and 9 co-immunoprecipitated with myc-PDK1 under baseline conditions and this interaction was stimulated by UCN-01; batimastat also markedly diminished this effect of the PDK1 inhibitor. Our results indicate that a series of protein kinase inhibitors, namely UCN-01 and OSU-03012 and to a lesser extent GSK2334470 and staurosporine induce PDK1 fragmentation and suggest that metalloproteinases could participate in this effect. PMID:25016091

Alcántara-Hernández, Rocío; Hernández-Méndez, Aurelio; Adolfo García-Sáinz, J

2014-10-01

159

A role for phosphoinositide 3-kinase in the completion of macropinocytosis and phagocytosis by macrophages  

PubMed Central

Phosphoinositide 3-kinase (PI 3-kinase) has been implicated in growth factor signal transduction and vesicular membrane traffic. It is thought to mediate the earliest steps leading from ligation of cell surface receptors to increased cell surface ruffling. We show here that inhibitors of PI 3-kinase inhibit endocytosis in macrophages, not by interfering with the initiation of the process but rather by preventing its completion. Consistent with earlier studies, the inhibitors wortmannin and LY294002 inhibited fluid-phase pinocytosis and Fc receptor-mediated phagocytosis, but they had little effect on the receptor-mediated endocytosis of diI-labeled, acetylated, low density lipoprotein. Large solute probes of endocytosis reported greater inhibition by wortmannin than smaller probes did, indicating that macropinocytosis was affected more than micropinocytosis. Since macropinocytosis and phagocytosis are actin-mediated processes, we expected that their inhibition by wortmannin resulted from deficient signaling from macrophage colony-stimulating factor (M-CSF) receptors or Fc receptors to the actin cytoskeleton. However, video microscopy showed cell surface ruffling in wortmannin-treated cells, and increased ruffling after addition of M-CSF or phorbol myristate acetate. Quantitative measurements of video data reported slightly diminished ruffling in wortmannin-treated cells. Remarkably, the ruffles that formed in wortmannin-treated macrophages all receded into the cytoplasm without closing into macropinosomes. Similarly, wortmannin and LY294002 did not inhibit the extension of actin-rich pseudopodia along IgG- opsonized sheep erythrocytes, but instead prevented them from closing into phagosomes. These findings indicate that PI 3-kinase is not necessary for receptor-mediated stimulation of pseudopod extension, but rather functions in the closure of macropinosomes and phagosomes into intracellular organelles. PMID:8947549

1996-01-01

160

IQGAP Proteins Reveal an Atypical Phosphoinositide (aPI) Binding Domain with a Pseudo C2 Domain Fold  

SciTech Connect

Class I phosphoinositide (PI) 3-kinases act through effector proteins whose 3-PI selectivity is mediated by a limited repertoire of structurally defined, lipid recognition domains. We describe here the lipid preferences and crystal structure of a new class of PI binding modules exemplified by select IQGAPs (IQ motif containing GTPase-activating proteins) known to coordinate cellular signaling events and cytoskeletal dynamics. This module is defined by a C-terminal 105-107 amino acid region of which IQGAP1 and -2, but not IQGAP3, binds preferentially to phosphatidylinositol 3,4,5-trisphosphate (PtdInsP3). The binding affinity for PtdInsP3, together with other, secondary target-recognition characteristics, are comparable with those of the pleckstrin homology domain of cytohesin-3 (general receptor for phosphoinositides 1), an established PtdInsP3 effector protein. Importantly, the IQGAP1 C-terminal domain and the cytohesin-3 pleckstrin homology domain, each tagged with enhanced green fluorescent protein, were both re-localized from the cytosol to the cell periphery following the activation of PI 3-kinase in Swiss 3T3 fibroblasts, consistent with their common, selective recognition of endogenous 3-PI(s). The crystal structure of the C-terminal IQGAP2 PI binding module reveals unexpected topological similarity to an integral fold of C2 domains, including a putative basic binding pocket. We propose that this module integrates select IQGAP proteins with PI 3-kinase signaling and constitutes a novel, atypical phosphoinositide binding domain that may represent the first of a larger group, each perhaps structurally unique but collectively dissimilar from the known PI recognition modules.

Dixon, Miles J.; Gray, Alexander; Schenning, Martijn; Agacan, Mark; Tempel, Wolfram; Tong, Yufeng; Nedyalkova, Lyudmila; Park, Hee-Won; Leslie, Nicholas R.; van Aalten, Daan M.F.; Downes, C. Peter; Batty, Ian H. (Toronto); (Dundee)

2012-10-16

161

Quantitative analysis of phosphoinositide 3-kinase (PI3K) signaling using live-cell total internal reflection fluorescence (TIRF) microscopy.  

PubMed

This unit focuses on the use of total internal reflection fluorescence (TIRF) microscopy and image analysis methods to study the dynamics of signal transduction mediated by class I phosphoinositide 3-kinases (PI3Ks) in mammalian cells. The first four protocols cover live-cell imaging experiments, image acquisition parameters, and basic image processing and segmentation. These methods are generally applicable to live-cell TIRF experiments. The remaining protocols outline more advanced image analysis methods, which were developed in our laboratory for the purpose of characterizing the spatiotemporal dynamics of PI3K signaling. These methods may be extended to analyze other cellular processes monitored using fluorescent biosensors. PMID:24510804

Johnson, Heath E; Haugh, Jason M

2013-01-01

162

Role of the phosphoinositide pathway in the light-dependent C4 phosphoenolpyruvate carboxylase phosphorylation cascade in Digitaria sanguinalis protoplasts.  

PubMed

Stimulus-response coupling in animal cells frequently involves the hydrolysis of PtdIns(4,5)P(2) which is catalysed by phosphoinositide-specific phospholipase C (PI-PLC). There is an increasing body of evidence for PI-PLC-based signalling in plant cells; however, the physiological role of this system remains poorly documented in plants. Our data provide the first evidence that a PI-PLC-based signalling system is a committed step in the transduction chain controlling the phosphorylation state of C(4) phosphoenolpyruvate carboxylase (PEPC), the regulation of which is central to the assimilation of atmospheric CO(2) in C(4) plants. PMID:11171220

Coursol, S; Pierre, J N; Vidal, J

2000-12-01

163

Effect of aging on alpha-1 adrenergic stimulation of phosphoinositide hydrolysis in various regions of rat brain  

SciTech Connect

The effects of aging were examined on the ability of alpha-1 adrenergic receptor agonists to stimulate phosphoinositide hydrolysis in three brain regions. Tissue minces of thalamus, cerebral cortex and hippocampus from 3-, 18- and 28-month-old male Fischer 344 rats were prelabeled with ({sup 3}H)myoinositol. Exposure of these prelabeled minces to phenylephrine and (-)-norepinephrine revealed that accumulation of ({sup 3}H)inositol phosphates was selectively reduced by 20 to 30% in the thalamus and cerebral cortex of the oldest age group. Analysis of concentration-response and competition binding curves indicated that this decrease was due to diminished agonist efficacy rather than diminished receptor affinity. The reduction in responsiveness to phenylephrine and (-)-norepinephrine in the cerebral cortex and the lack of any changes in the hippocampus parallel previously reported changes in the density of alpha-1 adrenergic receptors with aging. These data indicate that the ability of alpha-1 adrenergic receptor agonists to stimulate phosphoinositide hydrolysis is reduced in some, but not all, brain regions of aged Fischer 344 rats.

Burnett, D.M.; Bowyer, J.F.; Masserano, J.M.; Zahniser, N.R. (Univ. of Colorado Health Sciences Center, Denver (USA))

1990-12-01

164

Phylogenomics of phosphoinositide lipid kinases: perspectives on the evolution of second messenger signaling and drug discovery  

PubMed Central

Background Phosphoinositide lipid kinases (PIKs) generate specific phosphorylated variants of phosatidylinositols (PtdIns) that are critical for second messenger signaling and cellular membrane remodeling. Mammals have 19 PIK isoforms spread across three major families: the PtIns 3-kinases (PI3Ks), PtdIns 4-kinases (PI4Ks), and PtdIns-P (PIP) kinases (PIPKs). Other eukaryotes have fewer yet varying PIK complements. PIKs are also an important, emerging class of drug targets for many therapeutic areas including cancer, inflammatory and metabolic diseases and host-pathogen interactions. Here, we report the genomic occurrences and evolutionary relationships or phylogenomics of all three PIK families across major eukaryotic groups and suggest potential ramifications for drug discovery. Results Our analyses reveal four core eukaryotic PIKs which are type III PIK4A and PIK4B, and at least one homolog each from PI3K (possibly PIK3C3 as the ancestor) and PIP5K families. We also applied evolutionary analyses to PIK disease ontology and drug discovery. Mutated PIK3CA are known to be oncogenic and several inhibitors are in anti-cancer clinical trials. We found conservation of activating mutations of PIK3CA in paralogous isoforms suggesting specific functional constraints on these residues. By mapping published compound inhibition data (IC50s) onto a phylogeny of PI3Ks, type II PI4Ks and distantly related, MTOR, ATM, ATR and PRKDC kinases, we also show that compound polypharmacology corresponds to kinase evolutionary relationships. Finally, we extended the rationale for drugs targeting PIKs of malarial Plasmodium falciparum, and the parasites, Leishmania sp. and Trypanosoma sp. by identifying those PIKs highly divergent from human homologs. Conclusion Our phylogenomic analysis of PIKs provides new insights into the evolution of second messenger signaling. We postulate two waves of PIK diversification, the first in metazoans with a subsequent expansion in cold-blooded vertebrates that was post-emergence of Deutrostomia\\Chordata but prior to the appearance of mammals. Reconstruction of the evolutionary relationships among these lipid kinases also adds to our understanding of their roles in various diseases and assists in their development as potential drug targets. PMID:21208444

2011-01-01

165

Mechanical stress activates angiotensin II type 1 receptor without the involvement of angiotensin II  

Microsoft Academic Search

The angiotensin II type 1 (AT1) receptor has a crucial role in load-induced cardiac hypertrophy. Here we show that the AT1 receptor can be activated by mechanical stress through an angiotensin-II-independent mechanism. Without the involvement of angiotensin II, mechanical stress not only activates extracellular-signal-regulated kinases and increases phosphoinositide production in vitro, but also induces cardiac hypertrophy in vivo. Mechanical stretch

Yunzeng Zou; Hiroshi Akazawa; Yingjie Qin; Masanori Sano; Hiroyuki Takano; Tohru Minamino; Noriko Makita; Koji Iwanaga; Weidong Zhu; Sumiyo Kudoh; Haruhiro Toko; Koichi Tamura; Minoru Kihara; Toshio Nagai; Akiyoshi Fukamizu; Satoshi Umemura; Taroh Iiri; Toshiro Fujita; Issei Komuro

2004-01-01

166

Biphasic Modulation of Paracellular Claudin-5 Expression in Mouse Brain Endothelial Cells Is Mediated through the Phosphoinositide-3-Kinase/AKT Pathway.  

PubMed

Blood-brain barrier (BBB) integrity is compromised in many central nervous system disorders. Complex astrocyte and vascular endothelial cell interactions that regulate BBB integrity may be disturbed in these disorders. We previously showed that systemic administration of 3-chloropropanediol [(S)-(+)-3-chloro-1,2-propanediol] induces a transitory glial fibrillary acidic protein-astrocyte loss, reversible loss of tight junction complexes, and BBB integrity disruption. However, the intracellular signaling mechanisms that induce BBB integrity marker loss are unclear. We hypothesize that 3-chloropropanediol-induced modulation of tight junction protein expression is mediated through the phosphoinositide-3-kinase (PI3K)/AKT pathway. To test this hypothesis, we used a mouse brain endothelial cell line (bEnd.3) exposed to 3-chloropropanediol for up to 3 days. Results showed early reversible loss of sharp paracellular claudin-5 expression 90, 105, and 120 minutes after 3-chloropropanediol (500 ?M) treatment. Sharp paracellular claudin-5 profiles were later restored, but lost again by 2 and 3 days after 3-chloropropanediol treatment. Western blot and immunofluorescence studies showed increased p85-PI3K expression and transitory increased AKT (Thr308) phosphorylation at 15 and 30 minutes after 3-chloropropanediol administration. PI3K inhibitors LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one hydrochloride; 2.5-25 ?M] and PI-828 [2-(4-morpholinyl)-8-(4-aminopheny)l-4H-1-benzopyran-4-one; 0.1-10 ?M] prevented the 3-chloropropanediol-induced AKT (Thr308) phosphorylation and both early and late loss of paracellular claudin-5. However, AKT inhibitors only prevented the early changes in claudin-5 expression. This mechanistic study provides a greater understanding of the intracellular signaling pathways mediating tight junction protein expression and supports a hypothesis that two independent pathways triggered by PI3K mediate early and late loss of paracellular claudin-5 expression. PMID:25281324

Camire, Ryan B; Beaulac, Holly J; Brule, Stephanie A; McGregor, Annie I; Lauria, Emily E; Willis, Colin L

2014-12-01

167

RNA interference-mediated knockdown of Aurora-B alters the metastatic behavior of A549 cells via modulation of the phosphoinositide 3-kinase/Akt signaling pathway  

PubMed Central

Accumulating evidence has revealed that an elevated expression level of Aurora-B is associated with metastasis in various types of malignant tumor. However, it is currently unclear whether this molecule is involved in non-small lung cancer (NSCLC) metastasis, and the molecular mechanisms associated with Aurora-B and metastasis remain unknown. In the present study, in order to investigate whether Aurora-B is involved in the development and metastasis of NSCLC, the Aurora-B protein expression in NSCLC tissues was detected by immunohistochemistry and its association with metastasis was analyzed. The results revealed that the expression levels of the Aurora-B protein in tissues obtained from NSCLC patients with lymph node metastasis were significantly higher than those without metastatic disease. Furthermore, the effect of Aurora-B inhibition on A549 cell migration and invasion, as well as the activity of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway was evaluated. Aurora-B was inhibited in the A549 cells using short hairpin RNA, and the cell migration and invasion rates were investigated using wound healing and Transwell invasion assays. In addition, the expression of the main proteins in the PI3K/Akt/nuclear factor-?B (NF-?B) signaling pathway, and matrix metalloproteinase (MMP)-2 and -9 were measured by western blot analysis. The results demonstrated that cell migration and invasion were decreased as a result of silencing Aurora-B. Furthermore, the activity of the PI3K/Akt/NF-?B signaling pathway and the expression of MMP-2 and -9 protein were suppressed by silencing Aurora-B. The results of the present study indicate that the knockdown of Aurora-B suppresses A549 cell invasion and migration via the inhibition of the PI3K/Akt signaling pathway in vitro and thus, targeting Aurora-B may present a potential treatment strategy for NSCLC.

ZHOU, LONG DIAN; XIONG, XU; LONG, XIN HUA; LIU, ZHI LI; HUANG, SHAN HU; ZHANG, WEI

2014-01-01

168

Human conjunctival epithelial cells express histamine-1 receptors coupled to phosphoinositide turnover and intracellular calcium mobilization: role in ocular allergic and inflammatory diseases.  

PubMed

Dispase-dissociated primary cultures of human conjunctival epithelial (HCE) cells were stimulated with histamine and the generation of inositol phosphates ([3H]IPs) from [3H]phosphoinositide (PI) hydrolysis and the mobilization of intracellular calcium ([Ca2+]i) were studied using ion exchange chromatography and Fura-2 fluorescence techniques, respectively. Histamine (100 microM) maximally stimulated PI turnover in HCE cells by 210 +/- 10% (n = 21) above basal levels and with a potency (EC50) of 3.3 microM (n = 4). Histamine (EC50 = 5.8 microM, n = 3) rapidly mobilized [Ca2+]i which peaked within 10 sec but which was still significantly elevated 20 min after stimulation. The histamine-induced [Ca2+]i responses did not desensitize upon repeated applications of histamine. The effects of histamine (100 microM) on PI turnover and [Ca2+]i were potently antagonized by the H1-antagonists, emedastine (IC50 = 1.6-2.9 nM), triprolidine (IC50 = 3.1 nM) and levocabastine (IC50 = 8 nM), but weakly by the H2-(ranitidine/cimetidine) and H3-(thioperamide) antagonists (IC50s = 10-100 microM). In conclusion, HCE cells have been shown to possess functional H1-histamine receptors that couple to inositol phosphates generation which then mobilize intracellular calcium. These intracellular signaling mechanisms may be intimately linked with the process of inflammatory cytokine secretion from the HCE cells after stimulation by histamine released from the conjunctival mast cells. The current results strongly suggest that the HCE cells are active participants in mediating, and perhaps amplifying, the pro-inflammatory and allergic effects of histamine which is released from conjunctival mast cells during ocular allergic and inflammatory reactions. PMID:8983974

Sharif, N A; Xu, S X; Magnino, P E; Pang, I H

1996-08-01

169

Selective inhibition of the platelet phosphoinositide 3-kinase p110beta as promising new strategy for platelet protection during extracorporeal circulation.  

PubMed

Extracorporeal circulation (ECC) is used in cardiac surgery for cardiopulmonary bypass as well as in ventricular assist devices and for extracorporeal membrane oxygenation. Blood contact with the artificial surface and shear stress of ECC activates platelets and leukocytes resulting in a coagulopathy and proinflammatory events. Blockers of the platelet glycoprotein (GP) IIb/IIIa (CD41/CD61) can protect platelet function during ECC, a phenomenon called "platelet anaesthesia", but may be involved in post-ECC bleeding. We hypothesized that the new selective phosphoinositide 3-kinase p110beta inhibitor TGX-221 that inhibits shear-induced platelet activation without prolonging the bleeding time in vivo may also protect platelet function during ECC. Heparinized blood of healthy volunteers (n = 6) was treated in vitro with either the GP IIb/IIIa blocker tirofiban, TGX-221 or as control and circulated in an ECC model. Before and after 30 minutes circulation CD41 expression on the ECC-tubing as measure for platelet-ECC binding and generation of the platelet activation marker beta-thromboglobulin were determined using ELISA. Platelet aggregation and platelet-granulocyte binding were analysed in flow cytometry. After log-transforming the data statistical evaluation was performed using multifactor ANOVA in combination with Tukey's HSD test (global alpha = 5%). Tirofiban and TGX-221 inhibited platelet-ECC interaction, platelet aggregation and platelet-granulocyte binding. Tirofiban also inhibited ECC-induced beta-thromboglobulin release. The observed inhibition of platelet-ECC interaction and platelet activation by tirofiban contributes to explain the mechanism of "platelet anaesthesia". TGX-221 represents a promising alternative to GP IIb/IIIa blockade and should be further investigated for use during ECC in vivo. PMID:18327411

Straub, Andreas; Wendel, Hans Peter; Dietz, Klaus; Schiebold, Daniela; Peter, Karlheinz; Schoenwaelder, Simone M; Ziemer, Gerhard

2008-03-01

170

Studies on the metabolism of metallothionein and alkaline phosphatase of adult rat primary hepatocyte cultures: role of fetal calf serum and agonists of the phosphoinositide cascade  

Microsoft Academic Search

Summary Adult rat primary hepatocytes maintained in DMEM\\/F12 (Ham) media were used as a model system for studying the role of fetal calf serum (FCS) and agonists of the phosphoinositide cascade in the metabolism of metallothionein (MT) and alkaline phosphatase (ALP). Experiments were performed both after a 24 h preincubation with FCS and with bovine serum albumin (BSA). Hepatocytes were

K. Krfimer; A. Markwitan; J. Pallauf

1993-01-01

171

Inositol Phospholipid Metabolism in Arabidopsis. Characterized and Putative Isoforms of Inositol Phospholipid Kinase and Phosphoinositide-Specific Phospholipase C1  

PubMed Central

Phosphoinositides (PIs) constitute a minor fraction of total cellular lipids in all eukaryotic cells. They fulfill many important functions through interaction with a wide range of cellular proteins. Members of distinct inositol lipid kinase families catalyze the synthesis of these phospholipids from phosphatidylinositol. The hydrolysis of PIs involves phosphatases and isoforms of PI-specific phospholipase C. Although our knowledge of the roles played by plant PIs is clearly limited at present, there is no doubt that they are involved in many physiological processes during plant growth and development. In this review, we concentrate on inositol lipid-metabolizing enzymes from the model plant Arabidopsis for which biochemical characterization data are available, namely the inositol lipid kinases and PI-specific phospholipase Cs. The biochemical properties and structure of characterized and genome-predicted isoforms are presented and compared with those of the animal enzymes to show that the plant enzymes have some features clearly unique to this kingdom. PMID:12226484

Mueller-Roeber, Bernd; Pical, Christophe

2002-01-01

172

Dominant-Activating, Germline Mutations in Phosphoinositide 3-Kinase p110? Cause T Cell Senescence and Human Immunodeficiency  

PubMed Central

The p110? subunit of phosphoinositide 3-kinase (PI(3)K) is selectively expressed in leukocytes and is critical for lymphocyte biology. Here we report three different germline, heterozygous, gain-of-function mutations in the PIK3CD gene encoding p110? in fourteen patients from seven families. These patients presented with sinopulmonary infections, lymphadenopathy, nodular lymphoid hyperplasia and CMV and/or EBV viremia. Strikingly, naïve and central memory T cells were severely deficient, while senescent effector T cells were over-represented. In vitro, patient T cells exhibited increased phosphorylation of Akt and hyperactivation of mTOR, enhanced glucose uptake and terminal effector differentiation. Importantly, treatment with rapamycin to inhibit mTOR activity in vivo partially restored naïve T cells, largely rescued the in vitro T cell defects, and improved clinical course. PMID:24165795

Lucas, Carrie L.; Kuehn, Hye Sun; Zhao, Fang; Niemela, Julie E.; Deenick, Elissa K.; Palendira, Umaimainthan; Avery, Danielle T.; Moens, Leen; Cannons, Jennifer L.; Biancalana, Matthew; Stoddard, Jennifer; Ouyang, Weiming; Frucht, David L.; Rao, V. Koneti; Atkinson, T. Prescott; Agharahimi, Anahita; Hussey, Ashleigh A.; Folio, Les R.; Olivier, Kenneth N.; Fleisher, Thomas A.; Pittaluga, Stefania; Holland, Steven M.; Cohen, Jeffrey I.; Oliviera, Joao B.; Tangye, Stuart G.; Schwartzberg, Pamela L.; Lenardo, Michael J.; Uzel, Gulbu

2014-01-01

173

A Switch of G Protein-Coupled Receptor Binding Preference from Phosphoinositide 3-Kinase (PI3K)-p85 to Filamin A Negatively Controls the PI3K Pathway  

PubMed Central

Frequent oncogenic alterations occur in the phosphoinositide 3-kinase (PI3K) pathway, urging identification of novel negative controls. We previously reported an original mechanism for restraining PI3K activity, controlled by the somatostatin G protein-coupled receptor (GPCR) sst2 and involving a ligand-regulated interaction between sst2 with the PI3K regulatory p85 subunit. We here identify the scaffolding protein filamin A (FLNA) as a critical player regulating the dynamic of this complex. A preexisting sst2-p85 complex, which was shown to account for a significant basal PI3K activity in the absence of ligand, is disrupted upon sst2 activation. FLNA was here identified as a competitor of p85 for direct binding to two juxtaposed sites on sst2. Switching of GPCR binding preference from p85 toward FLNA is determined by changes in the tyrosine phosphorylation of p85- and FLNA-binding sites on sst2 upon activation. It results in the disruption of the sst2-p85 complex and the subsequent inhibition of PI3K. Knocking down FLNA expression, or abrogating FLNA recruitment to sst2, reversed the inhibition of PI3K and of tumor growth induced by sst2. Importantly, we report that this FLNA inhibitory control on PI3K can be generalized to another GPCR, the mu opioid receptor, thereby providing an unprecedented mechanism underlying GPCR-negative control on PI3K. PMID:22203038

Najib, Souad; Saint-Laurent, Nathalie; Esteve, Jean-Pierre; Schulz, Stefan; Boutet-Robinet, Elisa; Fourmy, Daniel; Lattig, Jens; Mollereau, Catherine; Pyronnet, Stephane; Susini, Christiane

2012-01-01

174

Limits for the detection of (poly-)phosphoinositides by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)  

Microsoft Academic Search

Matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been recently established as a powerful tool for the analysis of biomolecules. Here, MALDI-TOF MS was used for the detection of (poly-)phosphoinositides (PPI). PPI possess higher molecular weights than other phospholipids and a high phosphorylation-dependent negative charge. Both features affect the MALDI detection limits expressed as the minimum of

Matthias Müller; Jürgen Schiller; Marijana Petkovi?; Wolf Oehrl; Regina Heinze; Reinhard Wetzker; Klaus Arnold; Jürgen Arnhold

2001-01-01

175

Activation of phosphoinositide 3-kinase and Src family kinase is required for respiratory burst in rat neutrophils stimulated with artocarpol A  

Microsoft Academic Search

Artocarpol A (ART), a natural product isolated from Artocarpus rigida, stimulated superoxide anion (O2?) generation, which was inhibited by 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY 294002), a phosphoinositide 3-kinase (PI3K) inhibitor, in rat neutrophils. ART stimulated phosphorylation of protein kinase B (PKB\\/Akt) on both T308 and S473 residues, and LY 294002 inhibited these effects. Rat neutrophils expressed both class IA PI3K subunits (p85, p110?,

Yu-Hsiang Kuan; Ruey-Hseng Lin; Hui-Yi Lin; Li-Jiau Huang; Chi-Ren Tsai; Lo-Ti Tsao; Chun-Nan Lin; Ling-Chu Chang; Jih-Pyang Wang

2006-01-01

176

CAMK1 Phosphoinositide Signal-Mediated Protein Sorting and Transport Network in Human Hepatocellular Carcinoma (HCC) by Biocomputation.  

PubMed

We data-analyzed and constructed the high-expression CAMK1 phosphoinositide signal-mediated protein sorting and transport network in human hepatocellular carcinoma (HCC) compared with low-expression (fold change ?2) no-tumor hepatitis/cirrhotic tissues (HBV or HCV infection) in GEO data set, using integration of gene regulatory network inference method with gene ontology (GO). Our result showed that CAMK1 transport subnetwork upstream KCNQ3, LCN2, NKX2_5, NUP62, SORT1, STX1A activated CAMK1, and downstream CAMK1-activated AFP, ENAH, KPNA2, SLC4A3; CAMK1 signal subnetwork upstream BRCA1, DKK1, GPSM2, LEF1, NR5A1, NUP62, SORT1, SSTR5, TBL3 activated CAMK1, and downstream CAMK1-activated MAP2K6, SFRP4, SSTR5, TSHB, UBE2C in HCC. We proposed that CAMK1 activated network enhanced endosome to lysosome transport, endosome transport via multivesicular body sorting pathway, Golgi to endosome transport, intracellular protein transmembrane transport, intracellular protein transport, ion transport, mRNA transport, plasma membrane to endosome transport, potassium ion transport, protein transport, vesicle-mediated transport, anion transport, intracellular transport, androgen receptor signaling pathway, cell surface receptor-linked signal transduction, hormone-mediated signaling, induction of apoptosis by extracellular signals, signal transduction by p53 class mediator resulting in transcription of p21 class mediator, signal transduction resulting in induction of apoptosis, phosphoinositide-mediated signaling, Wnt receptor signaling pathway, as a result of inducing phosphoinositide signal-mediated protein sorting, and transport in HCC. Our hypothesis was verified by CAMK1 functional regulation subnetwork containing positive regulation of calcium ion transport via voltage gated calcium channel, cell proliferation, DNA repair, exocytosis, I-kappaB kinase/NF-kappaB cascade, immunoglobulin-mediated immune response, mast cell activation, natural killer cell-mediated cytotoxicity directed against tumor cell target, protein ubiquitination, sodium ion transport, survival gene product activity, T cell-mediated cytotoxicity, transcription, transcription from RNA polymerase II promoter, transcription initiation from RNA polymerase II promoter, transcription via serum response element binding, exit from mitosis, ubiquitin ligase activity during mitotic cell cycle, regulation of angiogenesis, apoptosis, cell growth, cell proliferation, cyclin-dependent protein kinase activity, gene expression, insulin secretion, steroid biosynthesis, transcription from RNA polymerase II promoter, transcription from RNA polymerase III promoter, cell cycle, cell migration, DNA recombination, and protein metabolism; also by CAMK1 negative functional regulation subnetwork including negative regulation of apoptosis, cell proliferation, centriole replication, fatty acid biosynthesis, lipoprotein lipase activity, MAPK activity, progression through cell cycle, transcription, transcription from RNA polymerase II promoter, cell growth, phosphorylation, and ubiquitin ligase activity during mitotic cell cycle in HCC. PMID:24825433

Wang, Lin; Huang, Juxiang; Jiang, Minghu; Chen, Qingchun; Jiang, Zhenfu; Feng, Haitao

2014-11-01

177

Inhibition of HIV-1 infection of peripheral blood mononuclear cells by a monoclonal antibody that binds to phosphoinositides and induces secretion of ?-chemokines.  

PubMed

A murine IgG mAb, WR321, selected for the ability to bind to phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-bisphosphate, but an inability to bind to any of 17 other lipids, including phosphatidylinositol, was examined as a probe for studying interactions of HIV-1 with primary human peripheral blood mononuclear cells. The WR321 mAb broadly neutralized CCR5-tropic strains of HIV-1 to prevent infection of the cells. The mAb also exhibited direct interaction with cells in the culture, resulting in secretion of chemokines that interfered with the interaction of HIV-1 virions with CCR5, the coreceptor for HIV-1 on the susceptible cells, leading to inhibition of infection by HIV-1. Phosphoinositides that are recognized by WR321 do not exist on the external surface of cells, but are concentrated on the inner surface (cytoplasmic leaflet) of the plasma membrane. Murine anti-phosphoinositide mAbs similar to WR321 have previously been directly microinjected into a variety of cultured cells, resulting in important changes in the functions of the cells. The present results suggest that binding of a mAb to phosphoinositides, resulting in secretion of ?-chemokines into the culture medium and neutralization of infection by CCR5-tropic HIV-1 of nearby susceptible cells, occurred by uptake and binding of the mAb at an intracellular location in the cultured cells that then led to secretion of HIV-1-inhibitory ?-chemokines. PMID:21040700

Matyas, Gary R; Wieczorek, Lindsay; Bansal, Divya; Chenine, Agnes-Laurence; Sanders-Buell, Eric; Tovanabutra, Sodsai; Kim, Jerome H; Polonis, Victoria; Alving, Carl R

2010-11-26

178

Roles of Phosphoinositides and of Spo14p (phospholipase D)-generated Phosphatidic Acid during Yeast Sporulation  

PubMed Central

During yeast sporulation, internal membrane synthesis ensures that each haploid nucleus is packaged into a spore. Prospore membrane formation requires Spo14p, a phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]-stimulated phospholipase D (PLD), which hydrolyzes phosphatidylcholine (PtdCho) to phosphatidic acid (PtdOH) and choline. We found that both meiosis and spore formation also require the phosphatidylinositol (PtdIns)/PtdCho transport protein Sec14p. Specific ablation of the PtdIns transport activity of Sec14p was sufficient to impair spore formation but not meiosis. Overexpression of Pik1p, a PtdIns 4-kinase, suppressed the sec14-1 meiosis and spore formation defects; conversely, pik1-ts diploids failed to undergo meiosis and spore formation. The PtdIns(4)P 5-kinase, Mss4p, also is essential for spore formation. Use of phosphoinositide-specific GFP-PH domain reporters confirmed that PtdIns(4,5)P2 is enriched in prospore membranes. sec14, pik1, and mss4 mutants displayed decreased Spo14p PLD activity, whereas absence of Spo14p did not affect phosphoinositide levels in vivo, suggesting that formation of PtdIns(4,5)P2 is important for Spo14p activity. Spo14p-generated PtdOH appears to have an essential role in sporulation, because treatment of cells with 1-butanol, which supports Spo14p-catalyzed PtdCho breakdown but leads to production of Cho and Ptd-butanol, blocks spore formation at concentrations where the inert isomer, 2-butanol, has little effect. Thus, rather than a role for PtdOH in stimulating PtdIns(4,5)P2 formation, our findings indicate that during sporulation, Spo14p-mediated PtdOH production functions downstream of Sec14p-, Pik1p-, and Mss4p-dependent PtdIns(4,5)P2 synthesis. PMID:14528019

Rudge, Simon A.; Sciorra, Vicki A.; Iwamoto, Michelle; Zhou, Chun; Strahl, Thomas; Morris, Andrew J.; Thorner, Jeremy; Engebrecht, JoAnne

2004-01-01

179

Regulation of the phosphoinositide pathway in cultured Sertoli cells from immature rats: effects of follicle-stimulating hormone and fluoride  

SciTech Connect

Many hormones elicit effects on target cells by stimulating the enzyme phospholipase-C, which catalyzes the hydrolysis of phosphoinositides to the intracellular second messengers diacylglycerol and inositol phosphates. The present study examined the roles of FSH and guanine nucleotide-binding proteins (G-proteins) in regulating the hydrolysis of phosphoinositides in Sertoli cells. Sertoli cell cultures prepared from 16- to 18-day-old rats were incubated for 24 h with myo-(2-3H) inositol to label endogenous phospholipids. Treatment of cells from 0.5-20 min with preparations of ovine FSH ranging in potency from 1-60 times that of NIH FSH S1 did not affect accumulation of inositol phosphates. Levels of total (3H)inositol phosphates ((3H)inositol mono-, di-, and triphosphates (IP, IP2, and IP3)) in FSH-treated cultures was 75-120% the levels in control cultures over the various time intervals studied. Addition of testosterone and the combination of testosterone plus retinoic acid, agents that have been shown to potentiate effects of FSH in other systems, did not affect accumulation of inositol phosphates in response to FSH. In contrast to the lack of effect on accumulation of inositol phosphates, FSH stimulated 4- to 11-fold increases in estradiol secretion over 24 h of culture, indicating that Sertoli cells were viable and responsive to FSH. AIF4- has been shown to activate G-proteins involved in regulation of adenylate cyclase activity. In the present study, AIF4- induced 4- to 5-fold increases in IP, IP2, and IP3 in experiments wherein FSH had no effect. Pretreatment of Sertoli cells with pertussis toxin (100 and 1000 ng/ml) for 24 h inhibited fluoride-induced generation of IP, IP2, and IP3 by 24-51%. Similar treatment with cholera toxin had no effect on basal or fluoride-induced generation of IP2 or IP3, but increased fluoride-induced generation of IP by 20-34%.

Quirk, S.M.; Reichert, L.E. Jr.

1988-07-01

180

Involvement of Class II Phosphoinositide 3-Kinase ?-Isoform in Antigen-Induced Degranulation in RBL-2H3 Cells  

PubMed Central

In this study, we present findings that suggest that PI3K-C2?, a member of the class II phosphoinositide 3-kinase (PI3K) subfamily, regulates the process of Fc?RI-triggered degranulation. RBL-2H3 cells were transfected with shRNA targeting PI3K-C2?. The knockdown impaired the Fc?RI-induced release of a lysosome enzyme, ?-hexosaminidase, without affecting the intracellular Ca2+ mobilization. The release of mRFP-tagged neuropeptide-Y, a reporter for the regulated exocytosis, was also decreased in the PI3K-C2?-deficient cells. The release was increased significantly by the expression of the siRNA-resistant version of PI3K-C2?. In wild-type cells, Fc?RI stimulation induced the formation of large vesicles, which were associated with CD63, a marker protein of secretory granules. On the vesicles, the existence of PI3K-C2? and PtdIns(3,4)P2 was observed. These results indicated that PI3K-C2? and its product PtdIns(3,4)P2 may play roles in the secretory process. PMID:25357130

Nigorikawa, Kiyomi; Hazeki, Kaoru; Guo, Ying; Hazeki, Osamu

2014-01-01

181

Propofol pretreatment attenuates lipopolysaccharide-induced acute lung injury in rats by activating the phosphoinositide-3-kinase/Akt pathway.  

PubMed

The aim of this study was to investigate the effect of propofol pretreatment on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the role of the phosphoinositide-3-kinase/protein kinase B (PI3K/Akt) pathway in this procedure. Survival was determined 48 h after LPS injection. At 1 h after LPS challenge, the lung wet- to dry-weight ratio was examined, and concentrations of protein, tumor necrosis factor-? (TNF-?), and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) were determined using the bicinchoninic acid method or ELISA. Lung injury was assayed via lung histological examination. PI3K and p-Akt expression levels in the lung tissue were determined by Western blotting. Propofol pretreatment prolonged survival, decreased the concentrations of protein, TNF-?, and IL-6 in BALF, attenuated ALI, and increased PI3K and p-Akt expression in the lung tissue of LPS-challenged rats, whereas treatment with wortmannin, a PI3K/Akt pathway specific inhibitor, blunted this effect. Our study indicates that propofol pretreatment attenuated LPS-induced ALI, partly by activation of the PI3K/Akt pathway. PMID:25387673

Zhao, L L; Hu, G C; Zhu, S S; Li, J F; Liu, G J

2014-12-01

182

Androgen receptor expression is regulated by the phosphoinositide 3-kinase/Akt pathway in normal and tumoral epithelial cells.  

PubMed Central

The androgen receptor (AR) is a ligand-responsive transcription factor known to play a central role in the pathogenesis of prostate cancer. However, the regulation of AR gene expression in the normal and pathological prostate remains poorly understood. This study focuses on the effect of the phosphoinositide 3-kinase (PI 3-kinase)/Akt axis on AR expression in vas deferens epithelial cells (VDEC), a suitable model to study androgen regulation of gene expression, and LNCaP cells (derived from a metastasis at the left supraclavicular lymph node from a 50-year-old patient with a confirmed diagnosis of metastatic prostate carcinoma). Taken together, our data show for the first time that the PI 3-kinase/Akt pathway is required for basal and dihydrotestosterone-induced AR protein expression in both VDEC and LNCaP. Inhibition of the PI 3-kinase/Akt pathway reduced AR expression and the decline in AR protein level correlated with a decrease in AR mRNA in VDEC but not in LNCaP. Since PI 3-kinase/Akt axis is active in prostate cancer, cross-talk between PI 3-kinase/Akt and AR signalling pathways may have implications for endocrine therapy. PMID:11971763

Manin, Michele; Baron, Silvere; Goossens, Karine; Beaudoin, Claude; Jean, Claude; Veyssiere, Georges; Verhoeven, Guido; Morel, Laurent

2002-01-01

183

Expression Analysis of a Stress-Related Phosphoinositide-Specific Phospholipase C Gene in Wheat (Triticum aestivum L.)  

PubMed Central

Plant phosphoinositide-specific phospholipases C (PI-PLCs) function in several essential plant processes associated with either development or environmental stress. In this report, we examined the expression patterns of TaPLC1 under drought and high salinity stress at the transcriptional and post-transcriptional levels. TaPLC1 mRNA was expressed in all wheat organs examined. U73122 and edelfosine, the PLC inhibitor, impaired seedling growth and enhanced seedling sensitivity to drought and high salinity stress. Though TaPLC1 expression in wheat was lowest at the seedling stage, it was strongly induced under conditions of stress. When 6-day-old wheat seedlings were treated with 200 mM NaCl or 20% (w/v) PEG 6000 for 6 or 12 h, respectively, the TaPLC1 transcript level increased by 16-fold compared to the control. Western blotting showed that the TaPLC protein concentration was also maintained at a high level from 24 to 48 h during stress treatment. Together, our results indicate the possible biological functions of TaPLC1 in regulating seedling growth and the response to drought and salinity stress. PMID:25121594

Wu, Lizhu; Hou, Mingyu; Dou, Shijuan; Pan, Yanyun

2014-01-01

184

PLC regulation: emerging pictures for molecular mechanisms.  

PubMed

Phosphoinositide-specific phospholipase C (PLC) enzymes are common signalling components linked to the activation of most cellular receptors. All PLC families are complex, modular, multi-domain proteins and together cover a broad spectrum of regulatory interactions, including direct binding to G protein subunits, small GTPases from Rho and Ras families, receptor and non-receptor tyrosine kinases and lipid components of cellular membranes. Recent structural determinations of PLC components and their complexes with regulatory proteins and direct mechanistic studies, together with earlier work, have provided the foundation to propose molecular mechanisms that stringently regulate PLC activity. PMID:20870410

Bunney, Tom D; Katan, Matilda

2011-02-01

185

TNF-alpha and leptin activate the alpha-isoform of class II phosphoinositide 3-kinase.  

PubMed

The class II PI 3-kinases are known to be activated by growth factors and chemokines but to date there are no reports of cytokine mediated regulation. Further, the intracellular signalling mechanisms regulating the class-II PI 3-kinases are poorly understood. We investigated the effects of the cytokines TNFalpha and leptin on the activity of the alpha isoform of the class II PI 3-kinase (PI3K-C2alpha) and find that these stimulate the enzyme 2-fold and 3-fold, in CHO cells and J774.2 macrophages, respectively. The stimulation by leptin was not accompanied by recruitment of any tyrosine phosphorylated proteins to PI3K-C2alpha and no shift in electrophoretic mobility was noted. Furthermore, we demonstrate that the actions of both cytokines are blocked by the MEK inhibitor PD98059. These findings indicate that the cytokines activate PI3K-C2alpha and do so by a mechanism that requires activation of the ERK pathway and thus differs from the mechanism used by insulin to activate the enzyme. PMID:12788079

Ktori, C; Shepherd, P R; O'Rourke, L

2003-06-20

186

Role of phosphoinositide 3-kinase beta in platelet aggregation and thromboxane A2 generation mediated by Gi signalling pathways.  

PubMed

PI3Ks (phosphoinositide 3-kinases) play a critical role in platelet functional responses. PI3Ks are activated upon P2Y12 receptor stimulation and generate pro-aggregatory signals. P2Y12 receptor has been shown to play a key role in the platelet aggregation and thromboxane A2 generation caused by co-stimulation with Gq or Gz, or super-stimulation of Gi pathways. In the present study, we evaluated the role of specific PI3K isoforms alpha, beta, gamma and delta in platelet aggregation, thromboxane A2 generation and ERK (extracellular-signal-regulated kinase) activation. Our results show that loss of the PI3K signal impaired the ability of ADP to induce platelet aggregation, ERK phosphorylation and thromboxane A2 generation. We also show that Gq plus Gi- or Gi plus Gz-mediated platelet aggregation, ERK phosphorylation and thromboxane A2 generation in human platelets was inhibited by TGX-221, a PI3Kbeta-selective inhibitor, but not by PIK75 (a PI3Kalpha inhibitor), AS252424 (a PI3Kgamma inhibitor) or IC87114 (a PI3Kdelta inhibitor). TGX-221 also showed a similar inhibitory effect on the Gi plus Gz-mediated platelet responses in platelets from P2Y1-/- mice. Finally, 2MeSADP (2-methyl-thio-ADP)-induced Akt phosphorylation was significantly inhibited in the presence of TGX-221, suggesting a critical role for PI3Kbeta in Gi-mediated signalling. Taken together, our results demonstrate that PI3Kbeta plays an important role in ADP-induced platelet aggregation. Moreover, PI3Kbeta mediates ADP-induced thromboxane A2 generation by regulating ERK phosphorylation. PMID:20441566

Garcia, Analia; Kim, Soochong; Bhavaraju, Kamala; Schoenwaelder, Simone M; Kunapuli, Satya P

2010-07-15

187

The Arabidopsis DREB2 genetic pathway is constitutively repressed by basal phosphoinositide-dependent phospholipase C coupled to diacylglycerol kinase  

PubMed Central

Phosphoinositide-dependent phospholipases C (PI-PLCs) are activated in response to various stimuli. They utilize substrates provided by type III-Phosphatidylinositol-4 kinases (PI4KIII) to produce inositol triphosphate and diacylglycerol (DAG) that is phosphorylated into phosphatidic acid (PA) by DAG-kinases (DGKs). The roles of PI4KIIIs, PI-PLCs, and DGKs in basal signaling are poorly understood. We investigated the control of gene expression by basal PI-PLC pathway in Arabidopsis thaliana suspension cells. A transcriptome-wide analysis allowed the identification of genes whose expression was altered by edelfosine, 30 ?M wortmannin, or R59022, inhibitors of PI-PLCs, PI4KIIIs, and DGKs, respectively. We found that a gene responsive to one of these molecules is more likely to be similarly regulated by the other two inhibitors. The common action of these agents is to inhibit PA formation, showing that basal PI-PLCs act, in part, on gene expression through their coupling to DGKs. Amongst the genes up-regulated in presence of the inhibitors, were some DREB2 genes, in suspension cells and in seedlings. The DREB2 genes encode transcription factors with major roles in responses to environmental stresses, including dehydration. They bind to C-repeat motifs, known as Drought-Responsive Elements that are indeed enriched in the promoters of genes up-regulated by PI-PLC pathway inhibitors. PA can also be produced by phospholipases D (PLDs). We show that the DREB2 genes that are up-regulated by PI-PLC inhibitors are positively or negatively regulated, or indifferent, to PLD basal activity. Our data show that the DREB2 genetic pathway is constitutively repressed in resting conditions and that DGK coupled to PI-PLC is active in this process, in suspension cells and seedlings. We discuss how this basal negative regulation of DREB2 genes is compatible with their stress-triggered positive regulation. PMID:23964284

Djafi, Nabila; Vergnolle, Chantal; Cantrel, Catherine; Wietrzynski, Wojciech; Delage, Elise; Cochet, Francoise; Puyaubert, Juliette; Soubigou-Taconnat, Ludivine; Gey, Delphine; Collin, Sylvie; Balzergue, Sandrine; Zachowski, Alain; Ruelland, Eric

2013-01-01

188

Non-redundant Roles of Phosphoinositide 3-Kinase Isoforms ? and ? in Glycoprotein VI-induced Platelet Signaling and Thrombus Formation*  

PubMed Central

Platelets are activated by adhesion to vascular collagen via the immunoglobulin receptor, glycoprotein VI (GPVI). This causes potent signaling toward activation of phospholipase C?2, which bears similarity to the signaling pathway evoked by T- and B-cell receptors. Phosphoinositide 3-kinase (PI3K) plays an important role in collagen-induced platelet activation, because this activity modulates the autocrine effects of secreted ADP. Here, we identified the PI3K isoforms directly downstream of GPVI in human and mouse platelets and determined their role in GPVI-dependent thrombus formation. The targeting of platelet PI3K? or -? strongly and selectively suppressed GPVI-induced Ca2+ mobilization and inositol 1,4,5-triphosphate production, thus demonstrating enhancement of phospholipase C?2 by PI3K?/?. That PI3K? and -? have a non-redundant function in GPVI-induced platelet activation and thrombus formation was concluded from measurements of: (i) serine phosphorylation of Akt, (ii) dense granule secretion, (iii) intracellular Ca2+ increases and surface expression of phosphatidylserine under flow, and (iv) thrombus formation, under conditions where PI3K?/? was blocked or p85? was deficient. In contrast, GPVI-induced platelet activation was insensitive to inhibition or deficiency of PI3K? or -?. Furthermore, PI3K?/?, but not PI3K?, contributed to GPVI-induced Rap1b activation and, surprisingly, also to Rap1b-independent platelet activation via GPVI. Together, these findings demonstrate that both PI3K? and -? isoforms are required for full GPVI-dependent platelet Ca2+ signaling and thrombus formation, partly independently of Rap1b. This provides a new mechanistic explanation for the anti-thrombotic effect of PI3K inhibition and makes PI3K? an interesting new target for anti-platelet therapy. PMID:19815551

Gilio, Karen; Munnix, Imke C. A.; Mangin, Pierre; Cosemans, Judith M. E. M.; Feijge, Marion A. H.; van der Meijden, Paola E. J.; Olieslagers, Serve; Chrzanowska-Wodnicka, Magdalena B.; Lillian, Rivka; Schoenwaelder, Simone; Koyasu, Shigeo; Sage, Stewart O.; Jackson, Shaun P.; Heemskerk, Johan W. M.

2009-01-01

189

Increasing Plasma Membrane Phosphatidylinositol(4,5)Bisphosphate Biosynthesis Increases Phosphoinositide Metabolism in Nicotiana tabacum[W][OA  

PubMed Central

A genetic approach was used to increase phosphatidylinositol(4,5)bisphosphate [PtdIns(4,5)P2] biosynthesis and test the hypothesis that PtdInsP kinase (PIPK) is flux limiting in the plant phosphoinositide (PI) pathway. Expressing human PIPKI? in tobacco (Nicotiana tabacum) cells increased plasma membrane PtdIns(4,5)P2 100-fold. In vivo studies revealed that the rate of 32Pi incorporation into whole-cell PtdIns(4,5)P2 increased >12-fold, and the ratio of [3H]PtdInsP2 to [3H]PtdInsP increased 6-fold, but PtdInsP levels did not decrease, indicating that PtdInsP biosynthesis was not limiting. Both [3H]inositol trisphosphate and [3H]inositol hexakisphosphate increased 3-and 1.5-fold, respectively, in the transgenic lines after 18 h of labeling. The inositol(1,4,5)trisphosphate [Ins(1,4,5)P3] binding assay showed that total cellular Ins(1,4,5)P3/g fresh weight was >40-fold higher in transgenic tobacco lines; however, even with this high steady state level of Ins(1,4,5)P3, the pathway was not saturated. Stimulating transgenic cells with hyperosmotic stress led to another 2-fold increase, suggesting that the transgenic cells were in a constant state of PI stimulation. Furthermore, expressing Hs PIPKI? increased sugar use and oxygen uptake. Our results demonstrate that PIPK is flux limiting and that this high rate of PI metabolism increased the energy demands in these cells. PMID:17496116

Im, Yang Ju; Perera, Imara Y.; Brglez, Irena; Davis, Amanda J.; Stevenson-Paulik, Jill; Phillippy, Brian Q.; Johannes, Eva; Allen, Nina S.; Boss, Wendy F.

2007-01-01

190

Propofol mediates signal transducer and activator of transcription 3 activation and crosstalk with phosphoinositide 3-kinase/AKT  

PubMed Central

We previously demonstrated that propofol, an intravenous anesthetic with anti-oxidative properties, activated the phosphoinositide 3-kinase (PI3K)/AKT pathway to increase the expression of B cell lymphoma (Bcl)-2 and, therefore the anti-apoptotic potential on cardiomyocytes. Here, we wanted to determine if propofol can also activate the Janus kinase (JAK) 2/signal transducer and activator of transcription (STAT) 3 pathway, another branch of cardioprotective signaling. The cellular response of nuclear factor kappa B (NF?B) and STAT3 was also evaluated. Cardiac H9c2 cells were treated by propofol alone or in combination with pretreatment by inhibitors for JAK2/STAT3 or PI3K/AKT pathway. STAT3 and AKT phosphorylation, and STAT3 translocation were measured by western blotting and immunofluorescence staining, respectively. Propofol treatment significantly increased STAT3 phosphorylation at both tyrosine 705 and serine 727 residues. Sustained early phosphorylation of STAT3 was observed with 25~75 ?M propofol at 10 and 30 min. Nuclear translocation of STAT3 was seen at 4 h after treatment with 50 ?M propofol. In cultured H9c2 cells, we further demonstrated that propofol-induced STAT3 phosphorylation was reduced by pretreatment with PI3K/AKT pathway inhibitors wortmannin or API-2. Conversely, pretreatment with JAK2/STAT3 pathway inhibitor AG490 or stattic inhibited propofol-induced AKT phosphorylation. In addition, propofol induced NF?B p65 subunit perinuclear translocation. Inhibition or knockdown of STAT3 was associated with increased levels of the NF?B p65 subunit. Our results suggest that propofol induces an adaptive response by dual activation and crosstalk of cytoprotective PI3K/AKT and JAK2/STAT3 pathways. Rationale to apply propofol clinically as a preemptive cardioprotectant during cardiac surgery is supported by our findings. PMID:25105067

Shravah, Jayant; Wang, Baohua; Pavlovic, Marijana; Kumar, Ujendra; Chen, David DY; Luo, Honglin; Ansley, David M

2014-01-01

191

Piperine inhibits type II phosphatidylinositol 4-kinases: a key component in phosphoinositides turnover.  

PubMed

Piperine has been shown to have anti-inflammatory properties. The molecular mechanisms by which it mediates anti-inflammatory activities remain elusive. Type II phosphatidylinositol 4-kinase(s) are key components in Fc?RI receptor-mediated signaling leading to inflammatory mediators release in RBL-2H3 cells. The effects of piperine on IgE-mediated signaling and mast cell degranulation were investigated. Pretreatment of RBL-2H3 cells with piperine inhibited IgE-induced activation of type II PtdIns 4-kinase(s). In vitro lipid kinase assays showed piperine-inhibited type II PtdIns 4-kinase activity in a dose-dependent fashion with no effect on PtdIns 3-kinase activity. Concomitantly, pretreatment of RBL-2H3 cells with piperine also inhibited IgE-induced ?-hexosaminidase release in RBL-2H3 cells. These results suggest that type II PtdIns 4-kinases are part of piperine-mediated anti-inflammatory signaling mechanisms. PMID:24671493

Bojjireddy, Naveen; Sinha, Ranjeet Kumar; Subrahmanyam, Gosukonda

2014-08-01

192

Distinct subpopulations in HaCaT cells as revealed by the characteristics of intracellular calcium release induced by phosphoinositide-coupled agonists  

Microsoft Academic Search

\\u000a Abstract Intracellular calcium release induced by transient applications of phosphoinositide agonists was measured using adherent\\u000a single HaCaT keratinocytes loaded with the acetoxymethyl derivative of fura-2. Application of ATP, bradykinin and formyl-Met-Leu-Phe\\u000a (fMLP) resulted in a transient increase in intracellular calcium concentration ([Ca2+]i) with an average half-width of 40 21 s and a decay time constant of 15 10 s (mean

Tamás Bíró; Imre Szabó; László Kovács; János Hunyadi; László Csernoch

1998-01-01

193

SNX-BAR proteins in phosphoinositide-mediated, tubular-based endosomal sorting.  

PubMed

The endocytic network is morphologically characterized by a wide variety of membrane bound compartments that are able to undergo dynamic re-modeling through tubular and vesicular structures. The precise molecular mechanisms governing such re-modeling, and the events that co-ordinated this with the major role of endosomes, cargo sorting, remain unclear. That said, recent work on a protein family of sorting nexins (SNX) - especially a subfamily of SNX that contain a BAR domain (SNX-BARs) - has begun to shed some much needed light on these issues and in particular the process of tubular-based endosomal sorting. SNX-BARs are evolutionary conserved in endosomal protein complexes such as retromer, where they co-ordinate membrane deformation with cargo selection. Furthermore a central theme emerges of SNX-BARs linking the forming membrane carrier to cytoskeletal elements for transport through motor proteins such as dynein. By studying these SNX-BARs, we are gaining an increasingly detailed appreciation of the mechanistic basis of endosomal sorting and how this highly dynamic process functions in health and disease. PMID:19914387

van Weering, Jan R T; Verkade, Paul; Cullen, Peter J

2010-06-01

194

PTEN Regulation, a Novel Function for the p85 Subunit of Phosphoinositide 3-Kinase  

NSDL National Science Digital Library

Timely regulation of phosphatidylinositol-3,4-bisphosphate [PI(3,4)P2] and phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3] abundance in cells is essential for the control of cellular homeostasis. The concentrations of these lipids are low in quiescent cells but rapidly and transiently increase following growth factor receptor (GFR) stimulation, which triggers cellular metabolic changes, proliferation, survival, and motility. Class IA phosphatidylinositol 3-kinase (PI3K), which is composed of a p85 (regulatory) and p110 (catalytic) subunits, is the enzyme generating PI(3,4)P2 and PI(3,4,5)P3 following GFR stimulation. Although the steps in GFR-induced activation of PI3K , are relatively well known, the mechanisms for subsequent 3-polyphospho-PI down-regulation are less understood. Examination of frequent genetic alterations in human cancer showed that PTEN (phosphatase with tensin homology on chromosome 10) is the major enzyme that decreases PI(3,4)P2 and PI(3,4,5)P3 cell content. Nonetheless, interpretation of the complexity of PTEN regulation remains a matter of debate. The recent description of diminished PTEN activity in liver-conditional knockout mice lacking the p85α PI3K regulatory subunit reveals a previously unknown p85α-dependent negative-feedback pathway that controls PI(3,4)P2 and PI(3,4,5)P3 half-life by regulating PTEN.

Domingo F. Barber (Universidad Autonoma de Madrid;Centro Nacional de Biotecnologia/Consejo Superiod de Investigaciones Cientificas REV); Maria Alvarado-Kristensson (Universidad Autonoma de Madrid;Centro Nacional de Biotecnologia/Consejo Superiod de Investigaciones Cientificas REV); Ana Gonzalez-Garcia (Universidad Autonoma de Madrid;Centro Nacional de Biotecnologia/Consejo Superiod de Investigaciones Cientificas REV); Rafael Pulido (Centro de Investigacion Principe Felipe; REV); Ana C. Carrera (Universidad Autonoma de Madrid;Centro Nacional de Biotecnologia/Consejo Superiod de Investigaciones Cientificas REV)

2006-11-21

195

Cholecystokinin receptors: disparity between phosphoinositide breakdown and amylase releasing activity of CCK analogues in pancreas  

SciTech Connect

Cholecystokinin (CCK) peptides are a family of hormones which also occur in brain. In pancreas CCK stimulates the release of amylase, a process that is dependent on the mobilization of intracellular Ca/sup 2 +/. Recent evidence suggests that inositol 1,4,5-trisphosphate, the breakdown product of phosphatidylinositol 4,5-bisphosphate, is responsible for the rise in intracellular Ca/sup 2 +/. Their laboratory has developed assays to study synthetic CCK analogues using radioligand binding, PI breakdown and amylase release. They have shown that there are good correlations among these three assay systems for the carboxy terminal fragments of CCK/sub 8/. Recently, they have discovered synthetic analogues of CCK/sub 4/ that are full agonists in amylase release but are ineffective in causing PI breakdown. In particular, A-61576, Boc-5-amino-2-indolemethylene-pent-2-ene-1-oyl-Leu-Asp-Phe-NH/sub 2/, is a full agonist in the amylase releasing assay, but is devoid of PI stimulating activity. A-61576 completely reverses the stimulation of PI response induced by CCK/sub 8/, indicative of an antagonist. Since a mechanism other than the PI breakdown is responsible for amylase release by A-61576, they suggest that separate receptors are responsible for PI breakdown and amylase release.

Lin, C.W.; Grant, D.; Bianchi, B.; Miller, T.; Witte, D.; Shue, Y.K.; Nadzan, A.

1986-03-05

196

?-Opioid Agonist Modulation of [3H]Thymidine Incorporation into DNA: Evidence for the Involvement of Pertussis Toxin-Sensitive G Protein-Coupled Phosphoinositide Turnover  

PubMed Central

A body of evidence has indicated that ?-opioid agonists can inhibit DNA synthesis in developing brain. We now report that ?-selective opioid agonists (U69593 and U50488) modulate [3H]thymidine incorporation into DNA in fetal rat brain cell aggregates in a dose- and developmental stage-dependent manner, ? agonists decreased thymidine incorporation by 35% in cultures grown for 7 days, and this process was reversed by the ?-selective antagonist, norbinaltorphimine, whereas in 21-day brain cell aggregates a 3.5-fold increase was evident. Cell labeling by [3H]thymidine was also inhibited by the ?-opioid agonist as shown by autoradiography. In addition, U69593 reduced basal rates of phosphoinositide formation in 7-day cultures and elevated it in 21-day cultures. Control levels were restored by norbinaltorphimine. Pertussis toxin blocked U69593-mediated inhibition of DNA synthesis. The action of ? agonists on thymidine incorporation in the presence of chelerythrine, a protein kinase C (PKC) inhibitor, or in combination with LiCl, a noncompetitive inhibitor of inositol phosphatase, was attenuated in both 7- and 21-day cultures. These results suggest that ? agonists may inhibit DNA synthesis via the phosphoinositide system with a pertussis toxin-sensitive G protein as transducer. In mixed glial cell aggregates, U50488 increased thymidine incorporation into DNA 3.1-fold, and this stimulation was reversed by the opioid antagonist naltrexone. PMID:8384252

Barg, Jacob; Belcheva, Mariana Mancheva; Rowinski, Jan; Coscia, Carmine James

2008-01-01

197

Src-family tyrosine kinases, phosphoinositide 3-kinase and Gab1 regulate extracellular signal-regulated kinase 1 activation induced by the type A endothelin-1 G-protein-coupled receptor.  

PubMed

The multisubstrate docking protein, growth-factor-receptor-bound protein 2-associated binder 1 (Gab1), which is phosphorylated on tyrosine residues following activation of receptor tyrosine kinases and cytokine receptors, regulates cell proliferation, survival and epithelial morphogenesis. Gab1 is also tyrosine phosphorylated following activation of G-protein-coupled receptors (GPCRs) where its function is poorly understood. To elucidate the role of Gab1 in GPCR signalling, we investigated the mechanism by which the type A endothelin-1 (ET-1) GPCR induced tyrosine phosphorylation of Gab1. Tyrosine phosphorylation of Gab1 induced by endothelin-1 was inhibited by PP1, a pharmacological inhibitor of Src-family tyrosine kinases. ET-1-induced Gab1 tyrosine phosphorylation was also inhibited by LY294002, which inhibits phosphoinositide 3-kinase (PI 3-kinase) enzymes. Inhibition of Src-family tyrosine kinases or PI 3-kinase also inhibited ET-1-induced activation of the mitogen activated protein kinase family member, extracellular signal-regulated kinase (ERK) 1. Thus we determined whether Gab1 regulated ET-1-induced ERK1 activation. Overexpression of wild-type Gab1 potentiated ET-1-induced activation of ERK1. Structure-function analyses of Gab1 indicated that mutant forms of Gab1 that do not bind the Src homology (SH) 2 domains of the p85 adapter subunit of PI 3-kinase or the SH2-domain-containing protein tyrosine phosphatase 2 (SHP-2) were impaired in their ability to potentiate ET-1-induced ERK1 activation. Taken together, our data indicate that PI 3-kinase and Src-family tyrosine kinases regulate ET-1-induced Gab1 tyrosine phosphorylation, which, in turn, induces ERK1 activation via PI 3-kinase- and SHP-2-dependent pathways. PMID:11695994

Bisotto, S; Fixman, E D

2001-11-15

198

Protein Phosphatase 2A (PP2A)-specific Ubiquitin Ligase MID1 Is a Sequence-dependent Regulator of Translation Efficiency Controlling 3-Phosphoinositide-dependent Protein Kinase-1 (PDPK-1)*  

PubMed Central

We have shown previously that the ubiquitin ligase MID1, mutations of which cause the midline malformation Opitz BBB/G syndrome (OS), serves as scaffold for a microtubule-associated protein complex that regulates protein phosphatase 2A (PP2A) activity in a ubiquitin-dependent manner. Here, we show that the MID1 protein complex associates with mRNAs via a purine-rich sequence motif called MIDAS (MID1 association sequence) and thereby increases stability and translational efficiency of these mRNAs. Strikingly, inclusion of multiple copies of the MIDAS motif into mammalian mRNAs increases production of the encoded proteins up to 20-fold. Mutated MID1, as found in OS patients, loses its influence on MIDAS-containing mRNAs, suggesting that the malformations in OS patients could be caused by failures in the regulation of cytoskeleton-bound protein translation. This is supported by the observation that the majority of mRNAs that carry MIDAS motifs is involved in developmental processes and/or energy homeostasis. Further analysis of one of the proteins encoded by a MIDAS-containing mRNA, namely PDPK-1 (3-phosphoinositide dependent protein kinase-1), which is an important regulator of mammalian target of rapamycin/PP2A signaling, showed that PDPK-1 protein synthesis is significantly reduced in cells from an OS patient compared with an age-matched control and can be rescued by functional MID1. Together, our data uncover a novel messenger ribonucleoprotein complex that regulates microtubule-associated protein translation. They suggest a novel mechanism underlying OS and point at an enormous potential of the MIDAS motif to increase the efficiency of biotechnological protein production in mammalian cells. PMID:21930711

Aranda-Orgilles, Beatriz; Rutschow, Desiree; Zeller, Raphael; Karagiannidis, Antonios I.; Kohler, Andrea; Chen, Changwei; Wilson, Timothy; Krause, Sven; Roepcke, Stefan; Lilley, David; Schneider, Rainer; Schweiger, Susann

2011-01-01

199

Modulation of phosphoinositide 3-kinase activation by cholesterol level suggests a novel positive role for lipid rafts in lysophosphatidic acid signalling.  

PubMed

Methyl-beta-cyclodextrin (MbetaCD) was used to explore a role for cholesterol-enriched plasma membrane microdomains in coupling lysophosphatidic acid (LPA) stimulation to phosphoinositide 3-kinase (PI3K) activation. Cholesterol depletion strongly inhibited the production of phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate in Vero cells stimulated with LPA. In agreement, the phosphorylation of Akt/protein kinase B, but not of Erk kinases, was suppressed by MbetaCD. MbetaCD did not interfere with the overall phospholipid metabolism, and its effects were reversed in cholesterol add-back experiments. Finally, PI3K was detected in lipid rafts prepared from control but not MbetaCD-treated cells, suggesting that these microdomains contribute to LPA signalling by compartmentalising component(s) of the PI3K pathway. PMID:12527380

Peres, Christine; Yart, Armelle; Perret, Bertrand; Salles, Jean Pierre; Raynal, Patrick

2003-01-16

200

Progress in the Preclinical Discovery and Clinical Development of Class I and Dual Class I/IV Phosphoinositide 3-Kinase (PI3K) Inhibitors  

PubMed Central

The phosphoinositide 3-kinases (PI3Ks) constitute an important family of lipid kinase enzymes that control a range of cellular processes through their regulation of a network of signal transduction pathways, and have emerged as important therapeutic targets in the context of cancer, inflammation and cardiovascular diseases. Since the mid-late 1990s, considerable progress has been made in the discovery and development of small molecule ATP-competitive PI3K inhibitors, a number of which have entered early phase human trials over recent years from which key clinical results are now being disclosed. This review summarizes progress made to date, primarily on the discovery and characterization of class I and dual class I/IV subtype inhibitors, together with advances that have been made in translational and clinical research, notably in cancer. PMID:21649578

Shuttleworth, S.J; Silva, F.A; Cecil, A.R.L; Tomassi, C.D; Hill, T.J; Raynaud, F.I; Clarke, P.A; Workman, P

2011-01-01

201

Synaptojanin 1 Mutation in Parkinson's Disease Brings Further Insight into the Neuropathological Mechanisms  

PubMed Central

Synaptojanin 1 (SYNJ1) is a phosphoinositide phosphatase highly expressed in nerve terminals. Its two phosphatase domains dephosphorylate phosphoinositides present in membranes, while its proline-rich domain directs protein-protein interactions with synaptic components, leading to efficient recycling of synaptic vesicles in neurons. Triplication of SYNJ1 in Down's syndrome is responsible for higher level of phosphoinositides, enlarged endosomes, and learning deficits. SYNJ1 downregulation in Alzheimer's disease models is protective towards amyloid-beta peptide (A?) toxicity. One missense mutation in one of SYNJ1 functional domains was recently incriminated in an autosomal recessive form of early-onset Parkinson's disease (PD). In the third decade of life, these patients develop progressive Parkinsonism with bradykinesia, dystonia, and variable atypical symptoms such as cognitive decline, seizures, and eyelid apraxia. The identification of this new gene, together with the fact that most of the known PD proteins play a role in synaptic vesicle recycling and lipid metabolism, points out that synaptic maintenance is a key player in PD pathological mechanisms. Studying PD genes as a network regulating synaptic activity could bring insight into understanding the neuropathological processes of PD and help identify new genes at fault in this devastating disorder.

Drouet, Valerie; Lesage, Suzanne

2014-01-01

202

Amplification of Chromosome 1q Genes Encoding the Phosphoinositide Signalling Enzymes PI4KB, AKT3, PIP5K1A and PI3KC2B in Breast Cancer  

PubMed Central

Little is known about the possible oncogenic roles of genes encoding for the phosphatidylinositol 4-kinases, a family of enzymes that regulate an early step in phosphoinositide signalling. To address this issue, the mutational status of all four human phosphatidylinositol 4-kinases genes was analyzed across 852 breast cancer samples using the COSMIC data resource. Point mutations in the phosphatidylinositol 4-kinase genes were uncommon and appeared in less than 1% of the patient samples however, 62% of the tumours had increases in gene copy number for PI4KB which encodes the phosphatidylinositol 4-kinase IIIbeta isozyme. Extending this analysis to subsequent enzymes in the phosphoinositide signalling cascades revealed that the only PIP5K1A, PI3KC2B and AKT3 genes exhibited similar patterns of gene copy number variation. By comparison, gene copy number increases for established oncogenes such as EGFR and HER2/Neu were only evident in 20% of the samples. The PI4KB, PIP5K1A, PI3KC2B and AKT3 genes are related in that they all localize to chromosome 1q which is often structurally and numerically abnormal in breast cancer. These results demonstrate that a gene quartet encoding a potential phosphoinositide signalling pathway is amplified in a subset of breast cancers. PMID:25368680

Waugh, Mark G.

2014-01-01

203

Modulatory effects of NMDA on phosphoinositide responses evoked by the metabotropic glutamate receptor agonist 1S,3R-ACPD in neonatal rat cerebral cortex.  

PubMed Central

1. The effect of NMDA-receptor stimulation on phosphoinositide signalling in response to the metabotropic glutamate receptor agonist 1-aminocyclopentane-1S,3R-dicarboxylic acid (1S,3R-ACPD) has been examined in neonatal rat cerebral cortex slices. 2. Total [3H]-inositol phosphate ([3H]-InsPx) accumulation, in the presence of 5 mM LiCl, in [3H]-inositol pre-labelled slices was concentration-dependently increased by 1S,3R-ACPD (EC50 16.6 microM) and, at a maximally effective concentration, 1S,3R-ACPD (300 microM) increased [3H]-InsPx accumulation by 12.8 fold over basal values. 3. [3H]-InsPx accumulation stimulated by 1S,1R-ACPD was enhanced by low concentrations of NMDA (3-30 microM), but not by higher concentrations (> 30 microM). [3H]-InsPx accumulations stimulated by 1S,3R-ACPD in the absence or presence of 10 microM NMDA were linear with time, at least over the 15 min period examined; however, in the presence of 100 microM NMDA the initial enhancement of 1S,3R-ACPD-stimulated phosphoinositide hydrolysis progressively decreased with time. 4. In the presence of a maximal enhancing concentration of NMDA (10 microM), the response to 1S,3R-ACPD (300 microM) was increased 1.9 fold and the EC50 for agonist-stimulated [3H]-InsPx accumulation decreased about 4 fold. The enhanced response to the metabotropic agonist was concentration-dependently inhibited by competitive and uncompetitive antagonists of NMDA-receptor activation. 5. 1S,3R-ACPD also stimulated inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) mass accumulation with an initial peak response (5-6 fold over basal) at 15 s decaying to a smaller (2 fold), but persistent elevated accumulation (1-10 min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7913380

Challiss, R. A.; Mistry, R.; Gray, D. W.; Nahorski, S. R.

1994-01-01

204

LY294002, an inhibitor of phosphoinositide 3-kinase given into rat hippocampus impairs acquisition, consolidation and retrieval of memory for one-trial step-down inhibitory avoidance.  

PubMed

Adult male Wistar rats were bilaterally implanted with indwelling cannulae in the CA1 region of the dorsal hippocampus. Once recovered from surgery, animals were submitted to one session of step-down inhibitory avoidance training (3.0 s, 0.4 mA footshock). Animals received a 0.5-microl infusion of saline, or of LY294002 (5, 50 or 500 microM), an inhibitor of the phosphoinositide 3-kinase (PI 3-K) family. Infusions were given 10 min before training, immediately post-training or 10 min prior to a 24-h retention test. In the pre- and post-training groups, the animals were tested twice: at 1.5 and 24 h after training, for short- (STM) and long-term memory (LTM), respectively. Pre- and post-training infusion of the drug inhibited both STM and LTM. Pre-test infusions impaired LTM retrieval. The effects can not be attributed to influences on locomotor, exploratory, pro- or anti-conflict behaviour, since LY294002 had no influence on elevated plus-maze behaviour. The results suggest that hippocampal PI 3-K is necessary for memory acquisition, consolidation and retrieval of the consolidation of step-down inhibitory avoidance in rats. This could be due to an interaction with the N-methyl-d-aspartate (NMDA) receptor complex or with activity of the extracellularly regulated protein kinase (ERK)-Ras signalling pathway. PMID:11856900

Barros, D M; Mello e Souza, T; de Souza, M M; Choi, H; DeDavid e Silva, T; Lenz, G; Medina, J H; Izquierdo, I

2001-12-01

205

p110gamma and p110delta phosphoinositide 3-kinase signaling pathways synergize to control development and functions of murine NK cells.  

PubMed

Phosphoinositide 3-kinases (PI-3Ks) are key enzymes for cell development, activation, and survival. Here we showed that PI-3K class IB and class IA catalytic subunits, p110gamma and p110delta, played a crucial role in the development and functions of murine NK cells. p110gamma deficiency and impairment of G protein-coupled receptor (GPRC) signaling prevented full NK cell maturation. Concomitant loss of p110gamma and p110delta exacerbated this defect, resulting in a very small population of NK cells with a highly immature phenotype in the bone marrow and periphery. Moreover, combined p110gamma and p110delta signals were required for cytotoxicity and activation of the kinase ERK during NK cell-target cell interaction. p110gamma played a major role in receptor-induced interferon-gamma (IFN-gamma) production through a pathway that involved the kinase ERK and 5-Lipoxigenase, which most likely generates lipid mediators activating GPRCs. Conversely, PI3Ks negatively regulated interleukin-12 (IL-12) and IL-18-induced IFN-gamma by modulating p38 kinase activation. Our data shed light on the multiple intersecting pathways through which PI3Ks control NK cell-mediated innate responses. PMID:17723215

Tassi, Ilaria; Cella, Marina; Gilfillan, Susan; Turnbull, Isaiah; Diacovo, Thomas G; Penninger, Josef M; Colonna, Marco

2007-08-01

206

T cell receptor-induced phosphoinositide-3-kinase p110? activity is required for T cell localization to antigenic tissue in mice  

PubMed Central

The establishment of T cell–mediated inflammation requires the migration of primed T lymphocytes from the blood stream and their retention in antigenic sites. While naive T lymphocyte recirculation in the lymph and blood is constitutively regulated and occurs in the absence of inflammation, the recruitment of primed T cells to nonlymphoid tissue and their retention at the site are enhanced by various inflammatory signals, including TCR engagement by antigen-displaying endothelium and resident antigen-presenting cells. In this study, we investigated whether signals downstream of TCR ligation mediated by the phosphoinositide-3-kinase (PI3K) subunit p110? contributed to the regulation of these events. T lymphocytes from mice expressing catalytically inactive p110? displayed normal constitutive trafficking and migratory responses to nonspecific stimuli. However, these cells lost susceptibility to TCR-induced migration and failed to localize efficiently to antigenic tissue. Importantly, we showed that antigen-induced T cell trafficking and subsequent inflammation was abrogated by selective pharmacological inhibition of PI3K p110? activity. These observations suggest that pharmacological targeting of p110? activity is a viable strategy for the therapy of T cell–mediated pathology. PMID:18259608

Jarmin, Sarah J.; David, Rachel; Ma, Liang; Chai, Jan-Guo; Dewchand, Hamlata; Takesono, Aya; Ridley, Anne J.; Okkenhaug, Klaus; Marelli-Berg, Federica M.

2008-01-01

207

Phosphatidic acid and phosphoinositides facilitate liposome association of Yas3p and potentiate derepression of ARE1 (alkane-responsive element one)-mediated transcription control.  

PubMed

In the n-alkane assimilating yeast Yarrowia lipolytica, the expression of ALK1, encoding a cytochrome P450 that catalyzes terminal mono-oxygenation of n-alkanes, is induced by n-alkanes. The transcription of ALK1 is regulated by a heterocomplex that comprises the basic helix-loop-helix transcription activators, Yas1p and Yas2p, and binds to alkane-responsive element 1 (ARE1) in the ALK1 promoter. An Opi1 family transcription repressor, Yas3p, represses transcription by binding to Yas2p. Yas3p localizes in the nucleus when Y. lipolytica is grown on glucose but localizes to the endoplasmic reticulum (ER) upon the addition of n-alkanes. In this study, we showed that recombinant Yas3p binds to the acidic phospholipids, phosphatidic acid (PA) and phosphoinositides (PIPs), in vitro. The ARE1-mediated transcription was enhanced in vivo in mutants defective in an ortholog of the Saccharomyces cerevisiae gene PAH1, encoding PA phosphatase, and in an ortholog of SAC1, encoding PIP phosphatase in the ER. Truncation mutation analyses for Yas3p revealed two regions that bound to PA and PIPs. These results suggest that the interaction with acidic phospholipids is important for the n-alkane-induced association of Yas3p with the ER membrane. PMID:24120453

Kobayashi, Satoshi; Hirakawa, Kiyoshi; Horiuchi, Hiroyuki; Fukuda, Ryouichi; Ohta, Akinori

2013-12-01

208

United at last: the tuberous sclerosis complex gene products connect the phosphoinositide 3-kinase/Akt pathway to mammalian target of rapamycin (mTOR) signalling.  

PubMed

The molecular interplay between the phosphoinositide 3-kinase (PI3K) pathway and mammalian target of rapamycin (mTOR) signalling in the control of cell growth and proliferation has been the subject of much interest and debate amongst cell biologists. A recent escalation of research in this area has come from the discovery of the tuberous sclerosis complex gene products, tuberin and hamartin, as central regulators of mTOR activation. The PI3K effector Akt/protein kinase B has been found to directly phosphorylate tuberin and is thereby thought to activate mTOR through inhibition of the tuberin-hamartin complex. The many recent studies aimed at defining the molecular nature of this revamped PI3K/Akt/mTOR pathway are reviewed here. The collective data discussed have laid the groundwork for important new insights into the many cancers caused by aberrant PI3K activation and the clinically challenging tuberous sclerosis complex disease and have suggested a possible means of treatment for both. PMID:12773158

Manning, B D; Cantley, L C

2003-06-01

209

Recruitment of OCRL and Inpp5B to phagosomes by Rab5 and APPL1 depletes phosphoinositides and attenuates Akt signaling  

PubMed Central

Sealing of phagosomes is accompanied by the disappearance of phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P2) from their cytoplasmic leaflet. Elimination of PtdIns(4,5)P2, which is required for actin remodeling during phagosome formation, has been attributed to hydrolysis by phospholipase C and phosphorylation by phosphatidylinositol 3-kinase. We found that two inositol 5-phosphatases, OCRL and Inpp5B, become associated with nascent phagosomes. Both phosphatases, which are Rab5 effectors, associate with the adaptor protein APPL1, which is recruited to the phagosomes by active Rab5. Knockdown of APPL1 or inhibition of Rab5 impairs association of OCRL and Inpp5B with phagosomes and prolongs the presence of PtdIns(4,5)P2 and actin on their membranes. Even though APPL1 can serve as an anchor for Akt, its depletion accentuated the activation of the kinase, likely by increasing the amount of PtdIns(4,5)P2 available to generate phosphatidylinositol (3,4,5)-trisphosphate. Thus, inositol 5-phosphatases are important contributors to the phosphoinositide remodeling and signaling that are pivotal for phagocytosis. PMID:22072788

Bohdanowicz, Michal; Balkin, Daniel M.; De Camilli, Pietro; Grinstein, Sergio

2012-01-01

210

Recruitment of OCRL and Inpp5B to phagosomes by Rab5 and APPL1 depletes phosphoinositides and attenuates Akt signaling.  

PubMed

Sealing of phagosomes is accompanied by the disappearance of phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P(2)) from their cytoplasmic leaflet. Elimination of PtdIns(4,5)P(2), which is required for actin remodeling during phagosome formation, has been attributed to hydrolysis by phospholipase C and phosphorylation by phosphatidylinositol 3-kinase. We found that two inositol 5-phosphatases, OCRL and Inpp5B, become associated with nascent phagosomes. Both phosphatases, which are Rab5 effectors, associate with the adaptor protein APPL1, which is recruited to the phagosomes by active Rab5. Knockdown of APPL1 or inhibition of Rab5 impairs association of OCRL and Inpp5B with phagosomes and prolongs the presence of PtdIns(4,5)P(2) and actin on their membranes. Even though APPL1 can serve as an anchor for Akt, its depletion accentuated the activation of the kinase, likely by increasing the amount of PtdIns(4,5)P(2) available to generate phosphatidylinositol (3,4,5)-trisphosphate. Thus, inositol 5-phosphatases are important contributors to the phosphoinositide remodeling and signaling that are pivotal for phagocytosis. PMID:22072788

Bohdanowicz, Michal; Balkin, Daniel M; De Camilli, Pietro; Grinstein, Sergio

2012-01-01

211

Blue light-induced chloroplast reorientations in Lemna trisulca L. (duckweed) are controlled by two separable cellular mechanisms as suggested by different sensitivity to wortmannin.  

PubMed

Chloroplast reorientations within mesophyll cells are among the most rapid physiological responses of higher plants to blue light. At light intensities below the saturation point of photosynthesis, chloroplasts move to the cell walls perpendicular to the direction of light and maximize light absorption (low-fluence rate response [LFR]). At light intensities above the saturation point of photosynthesis, chloroplasts redistribute to cell walls parallel to the direction of light (high-fluence rate response [HFR]). The actin-based mechanism is responsible for the light-induced chloroplast movements. We have found that an inhibitor of phosphoinositide-3-kinases, wortmannin, potently and irreversibly inhibited LFR and HFR chloroplast responses to blue light in Lemna trisulca L. mesophyll cells. Microscopic observations and photometric measurement indicated that 100 nM wortmannin specifically inhibited LFR in Lemna, whereas HFR displayed no sensitivity to the inhibitor at this concentration. A complete inhibition of the HFR could be obtained by 1 microM wortmannin. These data indicate that LFR is more sensitive to wortmannin than HFR and suggest that these two responses may be under the control of different cellular mechanisms. Our results suggest that phosphoinositide kinases and other phosphoinositide cycle enzymes may play a role in the transduction of the light signal to the actin cytoskeleton in Lemna as factors specifying the direction of chloroplast movements. A hypothetical model assuming three signaling pathways regulating light-induced chloroplast reorientations in mesophyll cells is proposed. PMID:15137511

Grabalska, Magdalena; Malec, Przemys?aw

2004-04-01

212

Thrombin-induced events in non-platelet cells are mediated by the unique proteolytic mechanism established for the cloned platelet thrombin receptor  

E-print Network

Abstract. We recently isolated a cDNA clone encoding a functional platelet thrombin receptor that defined a unique mechanism of receptor activation. Thrombin cleaves its receptor's extracellular amino terminal extension, unmasking a new amino terminus that functions as a tethered peptide ligand and activates the receptor. A novel peptide mimicking this new amino terminus was a full agonist for platelet secretion and aggregation, suggesting that this unusual mechanism accounts for platelet activation by thrombin. Does this mechanism also mediate thrombin's assorted actions on non-platelet cells? We now report that the novel thrombin receptor agonist peptide reproduces thrombininduced events (specifically, phosphoinositide hydrolysis and mitogenesis) in CCL-39 hamster lung fibroblasts,

David T. Hung; Thien-khaih Vu; T Nicolas A Nelken; Shaun R Coughlinfl

1992-01-01

213

Phosphoinositide 3-kinase Akt signaling pathway interacts with protein kinase C?2 in the regulation of physiologic developmental hypertrophy and heart function  

PubMed Central

The phosphoinositide 3-kinase (PI3-kinase)-protein kinase B (Akt) signaling pathway is essential in the induction of physiological cardiac hypertrophy. In contrast, protein kinase C ?2 (PKC?2) is implicated in the development of pathological cardiac hypertrophy and heart failure. Thus far, no clear association has been demonstrated between these two pathways. In this study, we examined the potential interaction between the PI3-kinase and PKC?2 pathways by crossing transgenic mice with cardiac specific expression of PKC?2, constitutively active (ca) PI3-kinase, and dominant-negative (dn) PI3-kinase. In caPI3-kinase/PKC?2 and dnPI3-kinase/PKC?2 double-transgenic mice, the heart weight-to-body weight ratios and cardiomyocyte sizes were similar to those observed in caPI3-kinase and dnPI3-kinase transgenic mice, respectively, suggesting that the regulation of physiological developmental hypertrophy via modulation of cardiomyocyte size proceeds through the PI3-kinase pathway. In addition, we observed that caPI3-kinase/PKC?2 mice showed improved cardiac function while the function of dnPI3-kinase/PKC?2 mice was similar to that of the PKC?2 group. PKC?2 protein levels in both dnPI3-kinase/PKC?2 and PKC?2 mice were significantly upregulated. Interestingly, however, PKC?2 protein expression was significantly attenuated in caPI3-kinase/PKC?2 mice. PI3-kinase activity measured by Akt phosphorylation was not affected by PKC?2 overexpression. These data suggest a potential interaction between these two pathways in the heart, where PI3-kinase is predominantly responsible for the regulation of physiological developmental hypertrophy and may act as an upstream modulator of PKC?2 with the potential for rescuing the pathological cardiac dysfunction induced by overexpression of PKC?. PMID:19122165

Rigor, Debra L.; Bodyak, Natalya; Bae, Soochan; Choi, Jun H.; Zhang, Li; Ter-Ovanesyan, Dmitry; He, Zhiheng; McMullen, Julie R.; Shioi, Tetsuo; Izumo, Seigo; King, George L.; Kang, Peter M.

2009-01-01

214

Overexpression of phospholipase D prevents actinomycin D-induced apoptosis through potentiation of phosphoinositide 3-kinase signalling pathways in Chinese-hamster ovary cells.  

PubMed

To examine the roles of PLD (phospholipase D) in the regulation of the apoptotic process, PLD1 and PLD2 were stably overexpressed in S1P3-CHO cells [CHO (Chinese-hamster ovary) cells expressing the S1P (sphingosine 1-phosphate) receptor S1P3]. Treatment of S1P3-CHO cells with ActD (actinomycin D) induced apoptosis, as shown by the occurrence of nuclear fragmentation and the caspase-dependent proteolytic cleavage of PARP [poly(ADP-ribose) polymerase] and protein kinase Cd. Overexpression of either PLD1 or PLD2 protected S1P3-CHO cells from ActD-induced apoptosis, as demonstrated by an increased number of viable cells and inhibition of PARP and protein kinase Cd cleavage. However, in the early phase of apoptosis, ActD induced an increase in PLD activity and activation of key factors in the cell-survival signalling pathways, such as PI3K (phosphoinositide 3-kinase), Akt, p70S6K (p70 S6 kinase) and ERK (extracellular-signal-regulated kinase). Furthermore, the ActD-induced activation of these survival signalling enzymes was potentiated by overexpression of either PLD1 or PLD2. The PI3K inhibitor LY294002 inhibited the ActD-induced activation of Akt and p70S6K, and completely abolished the effects of PLD1 or PLD2, whereas inhibition of ERK activity by the MEK inhibitor U0126 had a milder effect. The ActD-induced activation of p70S6K and ERKs was blocked by 1-butanol, but not by t-butanol; similar to S1P, exogenous PLD suppressed the ActD-induced events in the apoptosis signalling pathways. These results show that, in S1P3-CHO cells, increased expression of PLDs prevents ActD-induced apoptosis by enhanced activation of the PI3K signalling pathways. PMID:14640974

Yamada, Momoko; Banno, Yoshiko; Takuwa, Yoh; Koda, Masahiro; Hara, Akira; Nozawa, Yoshinori

2004-03-01

215

The phosphoinositide PI(3,5)P2 mediates activation of mammalian but not plant TPC proteins: functional expression of endolysosomal channels in yeast and plant cells.  

PubMed

Two-pore channel proteins (TPC) encode intracellular ion channels in both animals and plants. In mammalian cells, the two isoforms (TPC1 and TPC2) localize to the endo-lysosomal compartment, whereas the plant TPC1 protein is targeted to the membrane surrounding the large lytic vacuole. Although it is well established that plant TPC1 channels activate in a voltage- and calcium-dependent manner in vitro, there is still debate on their activation under physiological conditions. Likewise, the mode of animal TPC activation is heavily disputed between two camps favoring as activator either nicotinic acid adenine dinucleotide phosphate (NAADP) or the phosphoinositide PI(3,5)P2. Here, we investigated TPC current responses to either of these second messengers by whole-vacuole patch-clamp experiments on isolated vacuoles of Arabidopsis thaliana. After expression in mesophyll protoplasts from Arabidopsis tpc1 knock-out plants, we detected the Arabidopsis TPC1-EGFP and human TPC2-EGFP fusion proteins at the membrane of the large central vacuole. Bath (cytosolic) application of either NAADP or PI(3,5)P2 did not affect the voltage- and calcium-dependent characteristics of AtTPC1-EGFP. By contrast, PI(3,5)P2 elicited large sodium currents in hTPC2-EGFP-containing vacuoles, while NAADP had no such effect. Analogous results were obtained when PI(3,5)P2 was applied to hTPC2 expressed in baker's yeast giant vacuoles. Our results underscore the fundamental differences in the mode of current activation and ion selectivity between animal and plant TPC proteins and corroborate the PI(3,5)P2-mediated activation and Na(+) selectivity of mammalian TPC2. PMID:24770793

Boccaccio, Anna; Scholz-Starke, Joachim; Hamamoto, Shin; Larisch, Nina; Festa, Margherita; Gutla, Paul Vijay Kanth; Costa, Alex; Dietrich, Petra; Uozumi, Nobuyuki; Carpaneto, Armando

2014-11-01

216

Non-redundant roles of phosphoinositide 3-kinase isoforms alpha and beta in glycoprotein VI-induced platelet signaling and thrombus formation.  

PubMed

Platelets are activated by adhesion to vascular collagen via the immunoglobulin receptor, glycoprotein VI (GPVI). This causes potent signaling toward activation of phospholipase Cgamma2, which bears similarity to the signaling pathway evoked by T- and B-cell receptors. Phosphoinositide 3-kinase (PI3K) plays an important role in collagen-induced platelet activation, because this activity modulates the autocrine effects of secreted ADP. Here, we identified the PI3K isoforms directly downstream of GPVI in human and mouse platelets and determined their role in GPVI-dependent thrombus formation. The targeting of platelet PI3Kalpha or -beta strongly and selectively suppressed GPVI-induced Ca(2+) mobilization and inositol 1,4,5-triphosphate production, thus demonstrating enhancement of phospholipase Cgamma2 by PI3Kalpha/beta. That PI3Kalpha and -beta have a non-redundant function in GPVI-induced platelet activation and thrombus formation was concluded from measurements of: (i) serine phosphorylation of Akt, (ii) dense granule secretion, (iii) intracellular Ca(2+) increases and surface expression of phosphatidylserine under flow, and (iv) thrombus formation, under conditions where PI3Kalpha/beta was blocked or p85alpha was deficient. In contrast, GPVI-induced platelet activation was insensitive to inhibition or deficiency of PI3Kdelta or -gamma. Furthermore, PI3Kalpha/beta, but not PI3Kgamma, contributed to GPVI-induced Rap1b activation and, surprisingly, also to Rap1b-independent platelet activation via GPVI. Together, these findings demonstrate that both PI3Kalpha and -beta isoforms are required for full GPVI-dependent platelet Ca(2+) signaling and thrombus formation, partly independently of Rap1b. This provides a new mechanistic explanation for the anti-thrombotic effect of PI3K inhibition and makes PI3Kalpha an interesting new target for anti-platelet therapy. PMID:19815551

Gilio, Karen; Munnix, Imke C A; Mangin, Pierre; Cosemans, Judith M E M; Feijge, Marion A H; van der Meijden, Paola E J; Olieslagers, Servé; Chrzanowska-Wodnicka, Magdalena B; Lillian, Rivka; Schoenwaelder, Simone; Koyasu, Shigeo; Sage, Stewart O; Jackson, Shaun P; Heemskerk, Johan W M

2009-12-01

217

Inhibitory Effects of Isoquinoline Alkaloid Berberine on Ischemia-Induced Apoptosis via Activation of Phosphoinositide 3-Kinase/Protein Kinase B Signaling Pathway  

PubMed Central

Purpose Berberine is a type of isoquinoline alkaloid that has been used to treat various diseases. A neuroprotective effect of berberine against cerebral ischemia has been reported; however, the effects of berberine on apoptosis in relation to reactive astrogliosis and microglia activation under ischemic conditions have not yet been fully evaluated. In the present study, we investigated the effects of berberine on global ischemia-induced apoptosis, and focused on the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in the hippocampus using gerbils. Methods Gerbils received berberine orally once a day for 14 consecutive days, starting one day after surgery. In this study, a step-down avoidance task was used to assess short-term memory. Furthermore, we employed the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to evaluate DNA fragmentation, immunohistochemistry to investigate glial fibriallary acidic protein, CD11b, and caspase-3, and western blot to assess PI3K, Akt, Bax, Bcl-2, and cytochrome c. Results Our results revealed that berberine treatment alleviated ischemia-induced short-term memory impairment. Treatment with berbeine also attenuated ischemia-induced apoptosis and inhibited reactive astrogliosis and microglia activation. Furthermore, berberine enhanced phospho-PI3K and phospho-Akt expression in the hippocampus of ischemic gerbils. Conclusions Berberine exerted a neuroprotective effect against ischemic insult by inhibiting neuronal apoptosis via activation of the PI3K/Akt signaling pathway. The antiapoptotic effect of berberine was achieved through inhibition of reactive astrogliosis and microglia activation. Berberine may therefore serve as a therapeutic agent for stroke-induced neurourological problems. PMID:25279238

Kim, Mia; Shin, Mal Soon; Lee, Jae Min; Cho, Han Sam; Kim, Chang Ju; Kim, Young Joon; Choi, Hey Ran

2014-01-01

218

miR-375 Targets 3?-Phosphoinositide-Dependent Protein Kinase-1 and Regulates Glucose-Induced Biological Responses in Pancreatic ?-Cells  

PubMed Central

OBJECTIVE—MicroRNAs are short, noncoding RNAs that regulate gene expression. We hypothesized that the phosphatidylinositol 3-kinase (PI 3-kinase) cascade known to be important in ?-cell physiology could be regulated by microRNAs. Here, we focused on the pancreas-specific miR-375 as a potential regulator of its predicted target 3?-phosphoinositide–dependent protein kinase-1 (PDK1), and we analyzed its implication in the response of insulin-producing cells to elevation of glucose levels. RESEARCH DESIGN AND METHODS—We used insulinoma-1E cells to analyze the effects of miR-375 on PDK1 protein level and downstream signaling using Western blotting, glucose-induced insulin gene expression using quantitative RT-PCR, and DNA synthesis by measuring thymidine incorporation. Moreover, we analyzed the effect of glucose on miR-375 expression in both INS-1E cells and primary rat islets. Finally, miR-375 expression in isolated islets was analyzed in diabetic Goto-Kakizaki (GK) rats. RESULTS—We found that miR-375 directly targets PDK1 and reduces its protein level, resulting in decreased glucose-stimulatory action on insulin gene expression and DNA synthesis. Furthermore, glucose leads to a decrease in miR-375 precursor level and a concomitant increase in PDK1 protein. Importantly, regulation of miR-375 expression by glucose occurs in primary rat islets as well. Finally, miR-375 expression was found to be decreased in fed diabetic GK rat islets. CONCLUSIONS—Our findings provide evidence for a role of a pancreatic-specific microRNA, miR-375, in the regulation of PDK1, a key molecule in PI 3-kinase signaling in pancreatic ?-cells. The effects of glucose on miR-375 are compatible with the idea that miR-375 is involved in glucose regulation of insulin gene expression and ?-cell growth. PMID:18591395

El Ouaamari, Abdelfattah; Baroukh, Nadine; Martens, Geert A.; Lebrun, Patricia; Pipeleers, Daniel; van Obberghen, Emmanuel

2008-01-01

219

Inhibition by islet-activating protein, pertussis toxin, of P2-purinergic receptor-mediated iodide efflux and phosphoinositide turnover in FRTL-5 cells  

SciTech Connect

Exposure of FRTL-5 thyroid cells to ATP (1 microM to 1 mM) resulted in the stimulation of I- efflux in association with the induction of inositol trisphosphate production and intracellular Ca2+ mobilization. Nonhydrolyzable ATP derivatives, ADP and GTP, were also as effective in magnitude as ATP, whereas neither AMP nor adenosine exerted significant effect on I- efflux, suggesting a P2-purinergic receptor-mediated activation of I- efflux. Treatment of the cells with the islet-activating protein (IAP) pertussis toxin, which ADP-ribosylated a 41,000 mol wt membrane protein, effectively suppressed the phosphoinositide response to ATP in addition to ATP-dependent I- efflux at agonist concentrations below 10 microM. In contrast, the I- efflux stimulated by TSH, A23187, or phorbol myristate acetate was insusceptible to IAP. The IAP substrate, probably GTP-binding protein, is hence proposed to mediate the activation of P2-purinergic receptor-linked phospholipase-C in FRTL-5 cells. However, the responses to ATP, its nonhydrolyzable derivatives, or ADP at the higher agonist concentrations, especially above 100 microM, were only partially inhibited by IAP, even though the IAP substrate was totally ADP ribosylated by the toxin. The responses to GTP in the whole concentration range tested were not influenced by IAP treatment. Thus, signals arising from the P2-receptor might be transduced to phospholipase-C by two different pathways, i.e. IAP-sensitive and insensitive ones, and result in the stimulation of I- efflux.

Okajima, F.; Sho, K.; Kondo, Y.

1988-08-01

220

Arabidopsis thaliana Phosphoinositide-Specific Phospholipase C Isoform 3 (AtPLC3) and AtPLC9 have an Additive Effect on Thermotolerance.  

PubMed

The heat stress response is an important adaptation, enabling plants to survive challenging environmental conditions. Our previous work demonstrated that Arabidopsis thaliana Phosphoinositide-Specific Phospholipase C Isoform 9 (AtPLC9) plays an important role in thermotolerance. During prolonged heat treatment, mutants of AtPLC3 showed decreased heat resistance. We observed no obvious phenotypic differences between plc3 mutants and wild type (WT) seedlings under normal growth conditions, but after heat shock, the plc3 seedlings displayed a decline in thermotolerance compared with WT, and also showed a 40-50% decrease in survival rate and chlorophyll contents. Expression of AtPLC3 in plc3 mutants rescued the heat-sensitive phenotype; the AtPLC3-overexpressing lines also exhibited much higher heat resistance than WT and vector-only controls. The double mutants of plc3 and plc9 displayed increased sensitivity to heat stress, compared with either single mutant. In transgenic lines containing a AtPLC3:GUS promoter fusion, GUS staining showed that AtPLC3 expresses in all tissues, except anthers and young root tips. Using the Ca(2+)-sensitive fluorescent probe Fluo-3/AM and aequorin reconstitution, we showed that plc3 mutants show a reduction in the heat-induced Ca(2+) increase. The expression of HSP genes (HSP18.2, HSP25.3, HSP70-1 and HSP83) was down-regulated in plc3 mutants and up-regulated in AtPLC3-overexpressing lines after heat shock. These results indicated that AtPLC3 also plays a role in thermotolerance in Arabidopsis, and that AtPLC3 and AtPLC9 function additionally to each other. PMID:25149227

Gao, Kang; Liu, Yu-Liang; Li, Bing; Zhou, Ren-Gang; Sun, Da-Ye; Zheng, Shu-Zhi

2014-11-01

221

Platelet activation by bacterial phospholipase C involves phosphoinositide turnover and phosphorylation of 47,000 dalton but not 20,000 dalton protein  

SciTech Connect

This study was conducted to examine the role of phosphoinositides (PIns) and phosphorylation of 47,000 dalton (P47) and 20,000 dalton (P20) proteins in platelet activation by bacterial phospholipase C (PLC). PLC induced serotonin secretion (SS) and platelet aggregation (PA) in a concentration dependent manner. PLC (0.02 U/ml) caused phosphorylation of P47 in a time dependent manner (27% at 0.5 min to 378% at 7 min). PLC did not induce more than 15% phosphorylation of P20 by 7 min. Aspirin (500 ..mu..M) blocked phosphorylation of P20 but did not inhibit SS, PA or phosphorylation of P47. PLC (0.04 U/ml) decreased radioactivity (cpm) in /sup 32/P labeled phosphatidylinositol (PI), PI-4,5-bis-PO4 (PIP2) and PI-4-PO4 (PIP) by 20%, 12% and 7.5% respectively at 15 sec. The level of PI but not that of PIP2 returned to base line in 3 min. PIP level increased above control values within one min. PLC increased phosphatidic acid level (75% at 0.5 min. to 1545% at 3 min). In other experiments PLC produced diacylglycerol (DAG) in a time and concentration dependent manner. However, no DAG was detectable in the first 60 sec. These data suggest that: (a) PIns turnover and phosphorylation of P47 but not that of P20 is involved in platelet activation by PLC; and (b) DAG production from outer membrane phospholipids is not a prerequisite for platelet activation by PLC.

Huzoor-Akbar; Anwer, K.

1986-05-01

222

Dual Phosphoinositide 3-Kinase/Mammalian Target of Rapamycin Inhibitor NVP-BEZ235 Has a Therapeutic Potential and Sensitizes Cisplatin in Nasopharyngeal Carcinoma  

PubMed Central

Phosphoinositide 3-kinase (PI3K)/AKT/mammalian target of rapamycin inhibitor (mTOR) pathway is often constitutively activated in human tumor cells and thus has been considered as a promising drug target. To ascertain a therapeutical approach of nasopharyngeal carcinoma (NPC), we hypothesized NVP-BEZ235, a novel and potent imidazo[4,5-c] quinolone derivative, that dually inhibits both PI3K and mTOR kinases activities, had antitumor activity in NPC. Expectedly, we found that NVP-BEZ235 selectively inhibited proliferation of NPC cells rather than normal nasopharyngeal cells using MTT assay. In NPC cell lines, with the extended exposure, NVP-BEZ235 selectively inhibited proliferation of NPC cells harboring PIK3CA mutation, compared to cells with wild-type PIK3CA. Furthermore, exposure of NPC cells to NVP-BEZ235 resulted in G1 growth arrest by Propidium iodide uptake assay, reduction of cyclin D1and CDK4, and increased levels of P27 and P21 by Western blotting, but negligible apoptosis. Moreover, we found that cisplatin (CDDP) activated PI3K/AKT and mTORC1 pathways and NVP-BEZ235 alleviated the activation by CDDP through dually targeting PI3K and mTOR kinases. Also, NVP-BEZ235 combining with CDDP synergistically inhibited proliferation and induced apoptosis in NPC cells. In CNE2 and HONE1 nude mice xenograft models, orally NVP-BEZ235 efficiently attenuated tumor growth with no obvious toxicity. In combination with NVP-BEZ235 and CDDP, there was dramatic synergy in shrinking tumor volumes and inducing apoptosis through increasing Noxa, Bax and decreasing Mcl-1, Bcl-2. Based on the above results, NVP-BEZ235, which has entered phase I/II clinical trials in patients with advanced solid tumors, has a potential as a monotherapy or in combination with CDDP for NPC treatment. PMID:23533654

Deng, Rong; Wu, Xiao-Qi; Qin, Juan; Feng, Gong-Kan; Zhu, Xiao-Feng

2013-01-01

223

Hydrogen sulfide regulates Na+/H+ exchanger activity via stimulation of phosphoinositide 3-kinase/Akt and protein kinase G pathways.  

PubMed

Intracellular pH (pH(i)) is an important endogenous modulator of cardiac function. Inhibition of Na(+)/H(+) exchanger-1 (NHE-1) protects the heart by preventing Ca(2+) overload during ischemia/reperfusion. Hydrogen sulfide (H(2)S) has been reported to produce cardioprotection. The present study was designed to investigate the pH regulatory effect of H(2)S in rat cardiac myocytes and evaluate its contribution to cardioprotection. It was found that sodium hydrosulfide (NaHS), at a concentration range of 10 to 1000 ?M, produced sustained decreases in pH(i) in the rat myocytes in a concentration-dependent manner. NaHS also abolished the intracellular alkalinization caused by trans-(±)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl]benzeneacetamide methane-sulfonate hydrate (U50,488H), which activates NHEs. Moreover, when measured with an NHCl(4) prepulse method, NaHS was found to significantly suppress NHE-1 activity. Both NaHS and cariporide or [5-(2-methyl-5-fluorophenyl)furan-2-ylcarbonyl]guanidine (KR-32568), two NHE inhibitors, protected the myocytes against ischemia/reperfusion injury. However, coadministration of NaHS with KR-32568 did not produce any synergistic effect. Functional study showed that perfusion with NaHS significantly improved postischemic contractile function in isolated rat hearts subjected to ischemia/reperfusion. Blockade of phosphoinositide 3-kinase (PI3K) with 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), Akt with Akt VIII, or protein kinase G (PKG) with (9S,10R,12R)-2,3,9,10,11,12-hexahydro-10-methoxy-2,9-dimethyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4-i][1,6

Hu, Li-Fang; Li, Yu; Neo, Kay Li; Yong, Qian Chen; Lee, Shiau Wei; Tan, Benny Kwong Huat; Bian, Jin-Song

2011-11-01

224

Endosomal Targeting of the Phosphoinositide 3-Phosphatase MTMR2 Is Regulated by an N-terminal Phosphorylation Site*  

PubMed Central

MTMR2 is a member of the myotubularin family of inositol lipid phosphatases, a large protein-tyrosine phosphatase subgroup that is conserved from yeast to humans. Furthermore, the peripheral neuromuscular disease Charcot-Marie Tooth disease type 4B has been attributed to mutations in the mtmr2 gene. Because the molecular mechanisms regulating MTMR2 have been poorly defined, we investigated whether reversible phosphorylation might regulate MTMR2 function. We used mass spectrometry-based methods to identify a high stoichiometry phosphorylation site on serine 58 of MTMR2. Phosphorylation at Ser58, or a phosphomimetic S58E mutation, markedly decreased MTMR2 localization to endocytic vesicular structures. In contrast, a phosphorylation-deficient MTMR2 mutant (S58A) displayed constitutive localization to early endocytic structures. This localization pattern was accompanied by displacement of a PI(3)P-specific sensor protein and an increase in signal transduction pathways. Thus, MTMR2 phosphorylation is likely to be a critical mechanism by which MTMR2 access to its lipid substrate(s) is temporally and spatially regulated, thereby contributing to the control of downstream endosome maturation events. PMID:21372139

Franklin, Norah E.; Taylor, Gregory S.; Vacratsis, Panayiotis O.

2011-01-01

225

Phosphoinositide 3-kinase induced activation and cytoskeletal translocation of protein kinase CK2 in protease activated receptor 1-stimulated platelets.  

PubMed

CK2 is a highly conserved protein kinase involved in several cellular events. CK2 is expressed in platelets but its role in platelet activation remains poorly understood. In the present study, we tested the hypothesis that CK2 plays a role in platelet activation, particularly in the PAR1-dependent signal transduction pathway. The effect of CK2 and PI 3-kinase inhibitors on aggregation of platelets, activation of GPIIb/IIIa, activation and translocation of CK2 was examined. Platelets were incubated with the cell permeable CK2 inhibitors, DRB, DMAT and TBB and stimulated with the PAR1-AP (SFLLRNP). CK2 inhibitors showed the specific inhibitory pattern of platelet aggregation, characterized by a primary phase of aggregation followed by progressive disaggregation. CK2 inhibitors suppressed the activation of GPIIb/IIIa. PAR1-AP induced two-fold increase in CK2 activity and stimulated the translocation of CK2 from Triton X-100-soluble to -insoluble fraction. Preincubation of platelets with the PI 3-kinase inhibitor, wortmannin or LY294002, impaired PAR1-AP-induced aggregation of platelets. PAR1-AP-induced increase in CK2 activity and translocation of CK2 were inhibited by these treatments. Taken together, the present study demonstrated, for the first time, that PI 3-kinase-CK2 pathway plays an important role in the mechanism of PAR1-dependent platelet aggregation. PMID:21055793

Nakanishi, Kyoichi; Toyoda, Hidemi; Tanaka, Sigeki; Yamamoto, Hatsumi; Komada, Yoshihiro; Gabazza, Esteban C; Hayashi, Tatsuya; Suzuki, Koji; Ido, Masaru

2010-12-01

226

Phosphoinositide [PI(3,5)P2] lipid-dependent regulation of the general transcriptional regulator Tup1  

PubMed Central

Transcriptional activity of a gene is governed by transcriptional regulatory complexes that assemble/disassemble on the gene and control the chromatin architecture. How cytoplasmic components influence the assembly/disassembly of transcriptional regulatory complexes is poorly understood. Here we report that the budding yeast Saccharomyces cerevisiae has a chromatin architecture-modulating mechanism that is dependent on the endosomal lipid PI(3,5)P2. We identified Tup1 and Cti6 as new, highly specific PI(3,5)P2 interactors. Tup1—which associates with multiple transcriptional regulators, including the HDAC (histone deacetylase) and SAGA complexes—plays a crucial role in determining an activated or repressed chromatin state of numerous genes, including GAL1. We show that, in the context that the Gal4 activation pathway is compromised, PI(3,5)P2 plays an essential role in converting the Tup1-driven repressed chromatin structure into a SAGA-containing activated chromatin structure at the GAL1 promoter. Biochemical and cell biological experiments suggest that PI(3,5)P2 recruits Cti6 and the Cyc8–Tup1 corepressor complex to the late endosomal/vacuolar membrane and mediates the assembly of a Cti6–Cyc8–Tup1 coactivator complex that functions to recruit the SAGA complex to the GAL1 promoter. Our findings provide important insights toward understanding how the chromatin architecture and epigenetic status of a gene are regulated by cytoplasmic components. PMID:21536737

Han, Bong-Kwan; Emr, Scott D.

2011-01-01

227

Alpha 1-adrenergic receptor-mediated phosphoinositide hydrolysis and prostaglandin E2 formation in Madin-Darby canine kidney cells. Possible parallel activation of phospholipase C and phospholipase A2  

SciTech Connect

alpha 1-Adrenergic receptors mediate two effects on phospholipid metabolism in Madin-Darby canine kidney (MDCK-D1) cells: hydrolysis of phosphoinositides and arachidonic acid release with generation of prostaglandin E2 (PGE2). The similarity in concentration dependence for the agonist (-)-epinephrine in eliciting these two responses implies that they are mediated by a single population of alpha 1-adrenergic receptors. However, we find that the kinetics of the two responses are quite different, PGE2 production occurring more rapidly and transiently than the hydrolysis of phosphoinositides. The antibiotic neomycin selectively decreases alpha 1-receptor-mediated phosphatidylinositol 4,5-bisphosphate hydrolysis without decreasing alpha 1-receptor-mediated arachidonic acid release and PGE2 generation. In addition, receptor-mediated inositol trisphosphate formation is independent of extracellular calcium, whereas release of labeled arachidonic acid is largely calcium-dependent. Moreover, based on studies obtained with labeled arachidonic acid, receptor-mediated generation of arachidonic acid cannot be accounted for by breakdown of phosphatidylinositol monophosphate, phosphatidylinositol bisphosphate, or phosphatidic acid. Further studies indicate that epinephrine produces changes in formation or turnover of several classes of membrane phospholipids in MDCK cells. We conclude that alpha 1-adrenergic receptors in MDCK cells appear to regulate phospholipid metabolism by the parallel activation of phospholipase C and phospholipase A2. This parallel activation of phospholipases contrasts with models described in other systems which imply sequential activation of phospholipase C and diacylglycerol lipase or phospholipase A2.

Slivka, S.R.; Insel, P.A.

1987-03-25

228

Mechanism of cellular uptake of highly fluorescent conjugated polymer nanoparticles.  

PubMed

Conjugated polymer nanoparticles are formed by precipitation of highly fluorescent conjugated polymers to form small nanoparticles with extremely bright fluorescence. We characterized cellular uptake and cytotoxicity of 18 ± 5 nm PFBT conjugated polymer nanoparticles in J774A.1 cells. Significant nanoparticle uptake was observed, indicating efficient nanoparticle entry into cells, even for short (1 h) incubations. The high fluorescence of these nanoparticles allows extremely low loading concentrations; PFBT nanoparticle fluorescence in cells could be detected with loading concentrations of 155 pM (270 ppb). Cellular uptake slows at low temperature, consistent with endocytic entry. Nanoparticles colocalize with Texas Red dextran and are trafficked to lysosomes, as demonstrated by the location of nanoparticle fluorescence in perinuclear organelles that also stain with an anti-LAMP-1 antibody. Inhibition of uptake by phosphoinositide 3-kinase inhibitors implicates macropinocytosis as the operative endocytic mechanism. No significant cytotoxic or inflammatory effects could be observed, making PFBT nanoparticles attractive probes for live cell imaging. PMID:20863132

Fernando, Lawrence P; Kandel, Prakash K; Yu, Jiangbo; McNeill, Jason; Ackroyd, P Christine; Christensen, Kenneth A

2010-10-11

229

Physical process Mechanical mechanisms  

E-print Network

1 Physical process Generation · Mechanical mechanisms F = m·a · Electric/Magnetic mechanisms F = B·i·l · Fluid dynamic/Hydraulic mechanisms q, p, ij · Thermal/Optical #12;2 Source unit and source mechanisms ­ Monopoles......volume fluctuations ­ Dipoles ......pressure fluctuations

Berlin,Technische Universität

230

Arfs, phosphoinositides and membrane traffic.  

PubMed

Arf (ADP-ribosylation factor) GTP-binding proteins function in cells to regulate membrane traffic and structure. Arfs accomplish this task through modification of membrane lipids and the recruitment of proteins, including coat proteins and actin, to membrane surfaces. Arf1 and Arf6 are the most divergent and most studied human Arf proteins that localize predominantly to the Golgi complex and plasma membrane respectively. We have been studying the targeting of Arf1 and Arf6 to these specific compartments and the common and divergent activities that they exert on these membranes. We have found that Arf6 acts through activation of type I phosphatidylinositol 4-phosphate 5-kinases to generate phosphatidylinositol 4,5-bisphosphate and that this activity is instrumental in facilitating the actin cytoskeletal rearrangements and alterations in endosomal membrane trafficking observed with increased Arf6 activation. Arf1 can also stimulate the activity of phosphatidylinositol kinases and recruit coat proteins and actin cytoskeletal elements to the Golgi complex. PMID:16246097

Donaldson, J G

2005-12-01

231

MECHANICAL ENGINEERING What is Mechanical  

E-print Network

MECHANICAL ENGINEERING What is Mechanical Engineering? Mechanical engineering is one of the broadest engineering fields. Mechanical engineers are found in virtually all productive industries, from aircraft and automotive to consumer products and building equipment. In these jobs, mechanical engineers

232

An increase in phosphoinositide-specific phospholipase C activity precedes induction of C4 phosphoenolpyruvate carboxylase phosphorylation in illuminated and NH4Cl-treated protoplasts from Digitaria sanguinalis.  

PubMed

A Ca2+-dependent phosphoinositide-specific phospholipase C (PI-PLC) activity has been characterized in the microsomal fraction of Digitaria sanguinalis mesophyll cell protoplasts. Microsomal PI-PLC was found to be inhibited in vitro by a mammalian anti-PLC-delta1 antibody and by the aminosteroide U-73122, an inhibitor of PI-PLC activity in animal cells. In Western blot experiments, the antibody recognized an 85 kDa protein in both microsomal protein extracts from mesophyll protoplasts and rat brain protein extracts containing the authentic enzyme. The involvement of the microsomal PI-PLC in the light-dependent transduction pathway leading to the phosphorylation of C4 phosphoenolpyruvate carboxylase (PEPC) was investigated in D. sanguinalis protoplasts. A transient increase in the PI-PLC reaction product inositol-1,4,5-trisphosphate (Ins(1,4, 5)P3) was observed in situ during early induction of the C4 PEPC phosphorylation cascade. U-73122, but not the inactive analogue U-73343, efficiently blocked the transient accumulation of Ins(1,4, 5)P3, and both the increase in C4 PEPC kinase activity and C4 PEPC phosphorylation in illuminated and weak base-treated protoplasts. Taken together, these data suggest that PI-PLC-based signalling is a committed step in the cascade controlling the regulation of C4 PEPC phosphorylation in C4 leaves. PMID:10972876

Coursol, S; Giglioli-Guivarc'h, N; Vidal, J; Pierre, J N

2000-08-01

233

AMEAerospace & Mechanical  

E-print Network

AMEAerospace & Mechanical Engineering #12;Aerospace and Mechanical Engineers design complex mechanical, thermal, fluidic, acousti- cal, optical, and electronic systems, with char- acteristic sizes space. Aerospace and Mechanical Engineering (AME) students conduct basic and applied research within

Wang, Hai

234

Phosphoinositide-specific Phospholipase C ? 1b (PI-PLC?1b) Interactome: Affinity Purification-Mass Spectrometry Analysis of PI-PLC?1b with Nuclear Protein*  

PubMed Central

Two isoforms of inositide-dependent phospholipase C ?1 (PI-PLC?1) are generated by alternative splicing (PLC?1a and PLC?1b). Both isoforms are present within the nucleus, but in contrast to PLC?1a, the vast majority of PLC?1b is nuclear. In mouse erythroid leukemia cells, PI-PLC?1 is involved in the regulation of cell division and the balance between cell proliferation and differentiation. It has been demonstrated that nuclear localization is crucial for the enzymatic function of PI-PLC?1, although the mechanism by which this nuclear import occurs has never been fully characterized. The aim of this study was to characterize both the mechanism of nuclear localization and the molecular function of nuclear PI-PLC?1 by identifying its interactome in Friend's erythroleukemia isolated nuclei, utilizing a procedure that coupled immuno-affinity purification with tandem mass spectrometry analysis. Using this procedure, 160 proteins were demonstrated to be in association with PI-PLC?1b, some of which have been previously characterized, such as the splicing factor SRp20 (Srsf3) and Lamin B (Lmnb1). Co-immunoprecipitation analysis of selected proteins confirmed the data obtained via mass spectrometry. Of particular interest was the identification of the nuclear import proteins Kpna2, Kpna4, Kpnb1, Ran, and Rangap1, as well as factors involved in hematological malignancies and several anti-apoptotic proteins. These data give new insight into possible mechanisms of nuclear trafficking and functioning of this critical signaling molecule. PMID:23665500

Piazzi, Manuela; Blalock, William L.; Bavelloni, Alberto; Faenza, Irene; D'Angelo, Antonietta; Maraldi, Nadir M.; Cocco, Lucio

2013-01-01

235

Characterization of VPS34-IN1, a selective inhibitor of Vps34, reveals that the phosphatidylinositol 3-phosphate-binding SGK3 protein kinase is a downstream target of class III phosphoinositide 3-kinase  

PubMed Central

The Vps34 (vacuolar protein sorting 34) class III PI3K (phosphoinositide 3-kinase) phosphorylates PtdIns (phosphatidylinositol) at endosomal membranes to generate PtdIns(3)P that regulates membrane trafficking processes via its ability to recruit a subset of proteins possessing PtdIns(3)P-binding PX (phox homology) and FYVE domains. In the present study, we describe a highly selective and potent inhibitor of Vps34, termed VPS34-IN1, that inhibits Vps34 with 25 nM IC50 in vitro, but does not significantly inhibit the activity of 340 protein kinases or 25 lipid kinases tested that include all isoforms of class I as well as class II PI3Ks. Administration of VPS34-IN1 to cells induces a rapid dose-dependent dispersal of a specific PtdIns(3)P-binding probe from endosome membranes, within 1 min, without affecting the ability of class I PI3K to regulate Akt. Moreover, we explored whether SGK3 (serum- and glucocorticoid-regulated kinase-3), the only protein kinase known to interact specifically with PtdIns(3)P via its N-terminal PX domain, might be controlled by Vps34. Mutations disrupting PtdIns(3)P binding ablated SGK3 kinase activity by suppressing phosphorylation of the T-loop [PDK1 (phosphoinositide-dependent kinase 1) site] and hydrophobic motif (mammalian target of rapamycin site) residues. VPS34-IN1 induced a rapid ~50–60% loss of SGK3 phosphorylation within 1 min. VPS34-IN1 did not inhibit activity of the SGK2 isoform that does not possess a PtdIns(3)P-binding PX domain. Furthermore, class I PI3K inhibitors (GDC-0941 and BKM120) that do not inhibit Vps34 suppressed SGK3 activity by ~40%. Combining VPS34-IN1 and GDC-0941 reduced SGK3 activity ~80–90%. These data suggest SGK3 phosphorylation and hence activity is controlled by two pools of PtdIns(3)P. The first is produced through phosphorylation of PtdIns by Vps34 at the endosome. The second is due to the conversion of class I PI3K product, PtdIns(3,4,5)P3 into PtdIns(3)P, via the sequential actions of the PtdIns 5-phosphatases [SHIP1/2 (Src homology 2-domain-containing inositol phosphatase 1/2)] and PtdIns 4-phosphatase [INPP4B (inositol polyphosphate 4-phosphatase type II)]. VPS34-IN1 will be a useful probe to delineate physiological roles of the Vps34. Monitoring SGK3 phosphorylation and activity could be employed as a biomarker of Vps34 activity, in an analogous manner by which Akt is used to probe cellular class I PI3K activity. Combining class I (GDC-0941) and class III (VPS34-IN1) PI3K inhibitors could be used as a strategy to better analyse the roles and regulation of the elusive class II PI3K. PMID:25177796

Bago, Ruzica; Malik, Nazma; Munson, Michael J.; Prescott, Alan R.; Davies, Paul; Sommer, Eeva; Shpiro, Natalia; Ward, Richard; Cross, Darren; Ganley, Ian G.; Alessi, Dario R.

2014-01-01

236

Puzzling Mechanisms  

ERIC Educational Resources Information Center

The basis of a good mechanical puzzle is often a puzzling mechanism. This article will introduce some new puzzling mechanisms, like two knots that engage like gears, a chain whose links can be interchanged, and flat gears that do not come apart. It illustrates how puzzling mechanisms can be transformed into real mechanical puzzles, e.g., by…

van Deventer, M. Oskar

2009-01-01

237

Bohmian Mechanics  

E-print Network

Bohmian mechanics is a theory about point particles moving along trajectories. It has the property that in a world governed by Bohmian mechanics, observers see the same statistics for experimental results as predicted by quantum mechanics. Bohmian mechanics thus provides an explanation of quantum mechanics. Moreover, the Bohmian trajectories are defined in a non-conspiratorial way by a few simple laws.

Detlef Duerr; Sheldon Goldstein; Roderich Tumulka; Nino Zanghi

2009-03-15

238

Selective activation of the c-Jun N-terminal kinase (JNK) pathway fails to elicit Bax activation or apoptosis unless the phosphoinositide 3'-kinase (PI3K) pathway is inhibited.  

PubMed

c-Jun N-terminal kinase (JNK) is activated when cells are exposed to noxious stimuli. The role of JNK in apoptosis is subject to considerable debate; for example, JNK activation may promote or inhibit apoptosis depending on the cell type and stimulus involved. These conflicting results have arisen in part because few studies have successfully separated JNK activation from the primary stress-induced damage or from other stress-induced signalling pathways. Here we describe a conditional mutant, deltaMEKK1:ER*, which allows selective activation of the JNK cascade in the absence of any cellular stress. Activation of deltaMEKK1:ER* in CC139 fibroblasts resulted in the rapid and sustained activation of JNK without activating ERK or p38 or promoting IkappaBalpha phosphorylation. Activation of deltaMEKK1:ER* caused a reversible halt in cell growth but failed to induce apoptosis. In contrast, treatment of cells with LY294002, to inhibit phosphoinositide 3-kinase (PI3K), caused downregulation of Bcl-2 and Mcl-1 and allowed deltaMEKK1:ER* to elicit a robust apoptotic response characterized by activation of Bax and caspases. This PI3K-inhibitable, JNK-induced death response was not impeded, but actually accelerated, by cycloheximide. This suggests that JNK-induced activation of Bax and cell death does not require the upregulation of pro-death genes such as Bim or FasL, but rather proceeds through pre-existing components. However, if the PI3K cell survival pathway is not inhibited, even sustained activation of JNK exerts no overt proapoptotic effect in CC139 cells. PMID:12879014

Molton, Sarah A; Todd, Daniel E; Cook, Simon J

2003-07-24

239

The gene for a novel protein, a member of the protein disulphide isomerase/form I phosphoinositide-specific phospholipase C family, is amplified in hydroxyurea-resistant cells.  

PubMed Central

Cell lines selected in multiple steps for increasing resistance to hydroxyurea have been shown to have corresponding increases in ribonucleotide reductase activity. We have isolated a number of cDNA clones from a cDNA library constructed from a highly hydroxyurea-resistant hamster cell line, 600H, in which the activity of ribonucleotide reductase is elevated more than 80-fold. These clones correspond to genomic DNA sequences amplified in the 600H cell line compared with the V79 parental line. One of these cDNA clones, termed P5, codes for a 50 kDa protein detected by in vitro translation of poly(A)+ RNA isolated by hybridization/selection. The cDNA sequence contains a single open reading frame of 1317 nucleotides which encodes a polypeptide of 439 amino acids. The amino acid sequence deduced from the cDNA insert contains two copies of the 11-amino-acid sequence Val-Glu-Phe-Tyr-Ala-Pro-Trp-Cys-Gly-His-Cys. Duplicate copies of this sequence also occur in the active site of rat and human protein disulphide isomerase (also known as the beta-subunit of human prolyl 4-hydroxylase, tri-iodothyronine-binding protein) and in Form I phosphoinositide-specific phospholipase C, indicating that P5 falls into this newly defined superfamily of proteins. Genomic sequences similar to the cDNA clone are amplified 10-20-fold in hamster cells selected for resistance to increasing concentrations of hydroxyurea, a phenomenon observed earlier with cDNA clones for the M2 subunit of ribonucleotide reductase and ornithine decarboxylase. RNA blots probed with P5 cDNA show two poly(A)+ RNA species which are elevated in hydroxyurea-resistant cells. Images Fig. 1. Fig. 4. Fig. 5. Fig. 6. PMID:1311171

Chaudhuri, M M; Tonin, P N; Lewis, W H; Srinivasan, P R

1992-01-01

240

The Toll-like Receptor 9 Ligand, CpG Oligodeoxynucleotide, Attenuates Cardiac Dysfunction in Polymicrobial Sepsis, Involving Activation of Both Phosphoinositide 3 Kinase/Akt and Extracellular-Signal-Related Kinase Signaling  

PubMed Central

Background.?Toll-like receptors (TLRs) play a role in the pathophysiology of sepsis and multiple organ failure. This study examined the effect of CpG oligodeoxynucleotide (CpG-ODN), the TLR9 ligand, on polymicrobial sepsis–induced cardiac dysfunction. Methods.?Male C57BL/6 mice were treated with CpG-ODN, control CpG-ODN (control-ODN), or inhibitory CpG-ODN (iCpG-ODN) 1 hour prior to cecal ligation and puncture (CLP)–induced sepsis. Mice that underwent sham surgery served as sham controls. Cardiac function was examined by echocardiography before and 6 hours after CLP. Results.?Cardiac function was significantly decreased 6 hours after CLP. CpG-ODN prevented CLP-induced cardiac dysfunction, as evidenced by maintenance of the ejection fraction and fractional shortening. Control-ODN or iCpG-ODN did not alter CLP-induced cardiac dysfunction. CpG-ODN significantly attenuated CLP-induced myocardial apoptosis and increased myocardial Akt and extracellular-signal-related kinase (ERK) phosphorylation levels following CLP. In vitro experiments demonstrated that CpG-ODN promotes an association between TLR9 and Ras, resulting in Akt and ERK phosphorylation. Inhibition of phosphoinositide 3-kinase (PI3K) by Ly294002 or inhibition of ERK by U0126 in vivo abolished CpG-ODN attenuation of CLP-induced cardiac dysfunction. Conclusions.?CpG-ODN prevents CLP-induced cardiac dysfunction, in part through activation of PI3K/Akt and ERK signaling. Modulation of TLR9 could be an effective approach for treatment of cardiovascular dysfunction in patients with sepsis or septic shock. PMID:23359590

Gao, Ming; Ha, Tuanzhu; Zhang, Xia; Wang, Xiaohui; Liu, Li; Kalbfleisch, John; Singh, Krishna; Williams, David; Li, Chuanfu

2013-01-01

241

CD28 Costimulation: A Source of Vav-1 for TCR Signaling with the Help of SLP-76?  

NSDL National Science Digital Library

T cells require dual stimulation to become activated. When T cells encounter antigen-presenting cells, both the T cell receptor (TCR) and the CD28 coreceptor are ligated and activated. Michel and Acuto discuss how the adaptor SLP-76, which is recruited to the activated TCR complex, and the Rho family guanosine triphosphatase exchanger Vav-1, which is recruited by the CD28 receptor and TCR, may form a macromolecular complex that results in T cells activation. Vav-1 may serve as a central integrator between CD28 signaling and TCR signaling through its indirect effects on phosphoinositide 3-kinase-dependent signaling.

Frederique Michel (Institut Pasteur;Molecular Immunology Unit, Department of Immunology REV); Oreste Acuto (Institut Pasteur;Molecular Immunology Unit, Department of Immunology REV)

2002-08-06

242

Bohmian mechanics contradicts quantum mechanics  

E-print Network

Bohmian mechanics contradicts quantum mechanics Arnold Neumaier Institut fur Mathematik, Universit and quantum mechanics predict values of opposite sign for certain time correlations. The discrepancy can no loophole for claiming that Bohmian mechanics reproduces all predictions of quantum mechanics exactly

Neumaier, Arnold

243

Mechanical Systems  

NASA Technical Reports Server (NTRS)

The presentation provides an overview of requirement and interpretation letters, mechanical systems safety interpretation letter, design and verification provisions, and mechanical systems verification plan.

Davis, Robert E.

2002-01-01

244

Moving Forward: Mechanisms of Chemoattractant Gradient Sensing  

NSDL National Science Digital Library

Cells use an internal compass to sense the direction of chemoattractant gradients. This is used to bias pseudopod extension at the front of the cell and to orient cell polarization. Recent studies have highlighted the important roles played by phosphoinositide-3,4,5-triphosphate and small G proteins, but many questions remain.

PhD Jonathan Franca-Koh (Johns Hopkins University School of Medicine Department of Cell Biology); PhD Peter N. Devreotes (Johns Hopkins University School of Medicine Department of Cell Biology)

2004-10-01

245

Involvement of phospholipase D in insulin-like growth factor-I-induced activation of extracellular signal-regulated kinase, but not phosphoinositide 3-kinase or Akt, in Chinese hamster ovary cells.  

PubMed

Available evidence suggests the involvement of phospholipase D (PLD) in cell proliferation and survival. Phosphoinositide 3-kinase (PI 3-kinase)/Akt and extracellular signal-regulated kinases (ERKs) are signalling molecules that have essential roles in cell proliferation and survival. We previously demonstrated that sphingosine 1-phosphate (S1P)-induced PLD activation via the G-protein-coupled receptor endothelial differentiation gene (EDG) 3/S1P(3) was involved in S1P-induced stimulation of PI 3-kinase and Akt. In the present study, we examined the involvement of two PLD isozymes, PLD1 and PLD2, in insulin-like growth factor (IGF)-I receptor tyrosine kinase-mediated stimulation of PI 3-kinase/Akt and ERKs. IGF-I and to a lesser degree S1P stimulated PI 3-kinase activity in Chinese hamster ovary cells overexpressing EDG3/S1P(3). IGF-I-induced ERK phosphorylation was suppressed by butan-1-ol, but not butan-2-ol, whereas no effect of butanol was observed in IGF-I-induced Akt activation in S1P(3)-overexpressing Chinese hamster ovary cells. Overexpression of wild-type PLD1 and PLD2 substantially potentiated S1P-, but not IGF-I-, induced activation of PI 3-kinase and Akt, whereas overexpression of the catalytically inactive mutant of PLD1 or PLD2 did not affect the responses to either agonist. On the other hand, overexpression of wild-type PLD1 and PLD2 potentiated IGF-I- and, to much smaller extents, S1P-induced ERK stimulation. ERK activation by IGF-I as well as S1P was dependent on Ras, but Akt activation by IGF-I was not dependent on Ras. These results suggest that PLDs are involved in growth factor regulation of at least two signalling pathways, PI 3-kinase/Akt and ERKs, depending on the class of cell-surface receptors. PMID:12385647

Banno, Yoshiko; Takuwa, Yoh; Yamada, Momoko; Takuwa, Noriko; Ohguchi, Kenji; Hara, Akira; Nozawa, Yoshinori

2003-01-15

246

Classical Mechanics  

NSDL National Science Digital Library

This textbook, a standard in advanced classical mechanics, covers the breadth of the subject. This latest edition contains sections in contemporary classical mechanics, applications, and computer and mathematical techniques for solving systems.

Goldstein, Herbert; Poole Jr., Charles P.; Safko, John L.

2004-06-17

247

Geometric Mechanics  

Microsoft Academic Search

Mechanics for the nonmathematician-a modern approach For physicists, mechanics is quite obviously geometric, yet the classical approach typically emphasizes abstract, mathematical formalism. Setting out to make mechanics both accessible and interesting for nonmathematicians, Richard Talman uses geometric methods to reveal qualitative aspects of the theory. He introduces concepts from differential geometry, differential forms, and tensor analysis, then applies them to

Richard Talman

1999-01-01

248

Plasma membrane nanoporation as a possible mechanism behind infrared excitation of cells.  

PubMed

Objective. Short infrared (IR) laser pulses have been used to stimulate action potentials in neurons both in vivo and in vitro. However, the mechanism(s) underlying this phenomenon has remained elusive. In vitro studies have found that pulsed IR exposure generates a nearly instant change in capacitance in the plasma membrane, characterized by inward rectification, a common feature in pore-forming exposures, such as electrical pulses and acoustic shock waves. Based on this similarity, we hypothesize that the mechanism of IR stimulation is the formation of short-lived nanopores in the plasma membrane. These transient, small-diameter pores allow the influx of extracellular ions that lead to action potential generation, possibly through activation of secondary messenger pathways or depolarization of the cell membrane resulting in activation of voltage-gated ion channels. Approach. A variety of fluorescent markers are used to observe the cell response to IR stimulation to monitor for effects indicative of nanoporation in other modalities. Main results. We observe rapid, transient rises in intracellular Ca(2+), influx of YO-PRO-1 and propidium iodide into the cell signifying membrane permeabilization, cellular blebbing and swelling, and activation of the intracellular phosphoinositides lipid signaling pathway. Significance. This conclusion better explains the experimental observations and limitations of IR-induced neurological stimulation and represents a distinct theoretical shift in the understanding of the mechanism of IR-induced stimulation. PMID:25340253

Beier, Hope T; Tolstykh, Gleb P; Musick, Joshua D; Thomas, Robert J; Ibey, Bennett L

2014-12-01

249

Quantum Mechanics  

NASA Astrophysics Data System (ADS)

Preface; 1. Introduction; 2. Mathematical preliminaries; 3. The rules of quantum mechanics; 4. The connection between the fundamental rules and wave mechanics; 5. Further illustrations of the rules of quantum mechanics; 6. Further developments in one-dimensional wave mechanics; 7. The theory of angular momentum; 8. Wave mechanics in three dimensions: hydrogenic atoms; 9. Time-independent approximations for bound state problems; 10. Applications of static perturbation theory; 11. Identical particles; 12. Atomic structure; 13. Molecules; 14. The stability of matter; 15. Photons; 16. Interaction of non-relativistic charged particles and radiation; 17. Further topics in perturbation theory; 18. Scattering; 19. Special relativity and quantum mechanics: the Klein–Gordon equation; 20. The Dirac equation; 21. Interaction of a relativistic spin 1/2 particle with an external electromagnetic field; 22. The Dirac field; 23. Interaction between relativistic electrons, positrons, and photons; 24. The quantum mechanics of weak interactions; 25. The quantum measurement problem; Appendix A: useful inequalities for quantum mechanics; Appendix B: Bell's inequality; Appendix C: spin of the photon: vector spherical waves; Works cited; Bibliography; Index.

Commins, Eugene D.

2014-10-01

250

Fluid Mechanics  

Microsoft Academic Search

This text is intended for the study of fluid mechanics at an intermediate level. The presentation starts with basic concepts, in order to form a sound conceptual structure that can support engineering applications and encourage further learning. The presentation is exact, incorporating both the mathematics involved and the physics needed to understand the various phenomena in fluid mechanics. Where a

David Pnueli; Chaim Gutfinger

1997-01-01

251

Automotive Mechanics.  

ERIC Educational Resources Information Center

This curriculum guide, which was validated by vocational teachers and mechanics in the field, describes the competencies needed by entry-level automotive mechanics. This guide lists 15 competencies; for each competency, various tasks with their performance objective, student learning experiences, suggested instructional techniques, instructional…

Linder, Ralph C.; And Others

252

Geometric Mechanics  

NASA Astrophysics Data System (ADS)

Mechanics for the nonmathematician-a modern approach For physicists, mechanics is quite obviously geometric, yet the classical approach typically emphasizes abstract, mathematical formalism. Setting out to make mechanics both accessible and interesting for nonmathematicians, Richard Talman uses geometric methods to reveal qualitative aspects of the theory. He introduces concepts from differential geometry, differential forms, and tensor analysis, then applies them to areas of classical mechanics as well as other areas of physics, including optics, crystal diffraction, electromagnetism, relativity, and quantum mechanics. For easy reference, Dr. Talman treats separately Lagrangian, Hamiltonian, and Newtonian mechanics-exploring their geometric structure through vector fields, symplectic geometry, and gauge invariance respectively. Practical perturbative methods of approximation are also developed. Geometric Mechanics features illustrative examples and assumes only basic knowledge of Lagrangian mechanics. Of related interest . . . APPLIED DYNAMICS With Applications to Multibody and Mechatronic Systems Francis C. Moon A contemporary look at dynamics at an intermediate level, including nonlinear and chaotic dynamics. 1998 (0-471-13828-2) 504 pp. MATHEMATICAL PHYSICS Applied Mathematics for Scientists and Engineers Bruce Kusse and Erik Westwig A comprehensive treatment of the mathematical methods used to solve practical problems in physics and engineering. 1998 (0-471-15431-8) 680 pp.

Talman, Richard

1999-10-01

253

Mechanical memory  

DOEpatents

A first-in-first-out (FIFO) microelectromechanical memory apparatus (also termed a mechanical memory) is disclosed. The mechanical memory utilizes a plurality of memory cells, with each memory cell having a beam which can be bowed in either of two directions of curvature to indicate two different logic states for that memory cell. The memory cells can be arranged around a wheel which operates as a clocking actuator to serially shift data from one memory cell to the next. The mechanical memory can be formed using conventional surface micromachining, and can be formed as either a nonvolatile memory or as a volatile memory.

Gilkey, Jeffrey C. (Albuquerque, NM); Duesterhaus, Michelle A. (Albuquerque, NM); Peter, Frank J. (Albuquerque, NM); Renn, Rosemarie A. (Alburquerque, NM); Baker, Michael S. (Albuquerque, NM)

2006-08-15

254

Mechanical memory  

DOEpatents

A first-in-first-out (FIFO) microelectromechanical memory apparatus (also termed a mechanical memory) is disclosed. The mechanical memory utilizes a plurality of memory cells, with each memory cell having a beam which can be bowed in either of two directions of curvature to indicate two different logic states for that memory cell. The memory cells can be arranged around a wheel which operates as a clocking actuator to serially shift data from one memory cell to the next. The mechanical memory can be formed using conventional surface micromachining, and can be formed as either a nonvolatile memory or as a volatile memory.

Gilkey, Jeffrey C. (Albuquerque, NM); Duesterhaus, Michelle A. (Albuquerque, NM); Peter, Frank J. (Albuquerque, NM); Renn, Rosemarie A. (Albuquerque, NM); Baker, Michael S. (Albuquerque, NM)

2006-05-16

255

Mechanical Drives  

NSDL National Science Digital Library

This is a web page with learning objects on Mechanical Drives with lessons in: Belt & Chain Drives, and Shaft Couplings, Clutches & Brakes. Each lesson links to an interactive resource which helps to further explain the concepts.

2012-12-10

256

Fault mechanics  

SciTech Connect

Recent observational, experimental, and theoretical modeling studies of fault mechanics are discussed in a critical review of U.S. research from the period 1987-1990. Topics examined include interseismic strain accumulation, coseismic deformation, postseismic deformation, and the earthquake cycle; long-term deformation; fault friction and the instability mechanism; pore pressure and normal stress effects; instability models; strain measurements prior to earthquakes; stochastic modeling of earthquakes; and deep-focus earthquakes. Maps, graphs, and a comprehensive bibliography are provided. 220 refs.

Segall, P. (USAF, Geophysics Laboratory, Hanscom AFB, MA (United States))

1991-01-01

257

Statistical Mechanics  

Microsoft Academic Search

We review and further develop a mathematical framework for non-equilibrium quantum statistical mechanics recently proposed in (JP4, JP5, JP6, Ru3, Ru4, Ru5, Ru6). In the alge- braic formalism of quantum statistical mechanics we introduce notions of non-equilibrium steady states, entropy production and heat fluxes, and study their properties. Our basic paradigm is a model of a small (finite) quantum system

V. Jaksi ´; C.-A. Pillet

1937-01-01

258

Fluid Mechanics  

NASA Astrophysics Data System (ADS)

This text is intended for the study of fluid mechanics at an intermediate level. The presentation starts with basic concepts, in order to form a sound conceptual structure that can support engineering applications and encourage further learning. The presentation is exact, incorporating both the mathematics involved and the physics needed to understand the various phenomena in fluid mechanics. Where a didactical choice must be made between the two, the physics prevails. Throughout the book the authors have tried to reach a balance between exact presentation, intuitive grasp of new ideas, and creative applications of concepts. This approach is reflected in the examples presented in the text and in the exercises given at the end of each chapter. Subjects treated are hydrostatics, viscous flow, similitude and order of magnitude, creeping flow, potential flow, boundary layer flow, turbulent flow, compressible flow, and non-Newtonian flows. This book is ideal for advanced undergraduate students in mechanical, chemical, aerospace, and civil engineering. Solutions manual available.

Pnueli, David; Gutfinger, Chaim

1997-01-01

259

Cellular Mechanisms of Gravitropic Response in Higher Plants  

NASA Astrophysics Data System (ADS)

The evolutionary success of land plants in adaptation to the vectorial environmental factors was based mainly on the development of polarity systems. In result, normal plant ontogenesis is based on the positional information. Polarity is a tool by which the developing plant organs and tissues are mapped and the specific three-dimensional structure of the organism is created. It is due to their polar organization plants are able to orient themselves relative to the gravity vector and different vectorial cues, and to respond adequately to various stimuli. Gravitation is one of the most important polarized environmental factor that guides the development of plant organisms in space. Every plant can "estimate" its position relative to the gravity vector and correct it, if necessary, by means of polarized growth. The direction and the magnitude of gravitational stimulus are constant during the whole plant ontogenesis. The key plant response to the action of gravity is gravitropism, i.e. the directed growth of organs with respect to the gravity vector. This response is a very convenient model to study the mechanisms of plant orientation in space. The present report is focused on the main cellular mechanisms responsible for graviropic bending in higher plants. These mechanisms and structures include electric polarization of plant cells, Ca ({2+) }gradients, cytoskeleton, G-proteins, phosphoinositides and the machinery responsible for asymmetric auxin distribution. Those mechanisms tightly interact demonstrating some hierarchy and multiple feedbacks. The Ca (2+) gradients provide the primary physiological basis of polarity in plant cells. Calcium ions influence on the bioelectric potentials, the organization of actin cytoskeleton, the activity of Ca (2+) -binding proteins and Ca (2+) -dependent protein kinases. Protein kinases modulate transcription factors activity thereby regulating the gene expression and switching the developmental programs. Actin cytoskeleton affects the molecular machinery of polar auxin transport. It results in the changes of auxin gradients in plant organs and tissues, which modulate all cellular mechanisms of polarity via multiple feedback loops. The understanding of the mechanisms of plant organism orientation relative to the gravity vector will allow us to develop efficient technologies for plant growing in microgravity conditions at orbital space stations and during long piloted space flights. This work was supported by the grant of Russian Foundation for Basic Research (N 14-04-01-624) and by the grant of St.-Petersburg State University (N 1.38.233.2014).

Medvedev, Sergei; Smolikova, Galina; Pozhvanov, Gregory; Suslov, Dmitry

260

Quantum Mechanics  

NSDL National Science Digital Library

This website contains a number of descriptions of quantum mechanical phenomena, using 3D animations to illustrate the physics. The goal is to introduce basic concepts and phenomena using simulations rather than complex mathematics. The time-dependence of quantum systems is a focus of this material.

De Raedt, Hans; Michielsen, Kristel

2010-03-25

261

& Mechanical Engineering  

E-print Network

knowledge about the earth's oceans and atmospheres. Other e orts advance our understanding of such areas from microns to tens of kilometers. Such systems are used everywhere, from the depths of the ocean and far underground, to near-Earth, planetary, interplanetary and galactic space. Aerospace and Mechanical

Zhou, Chongwu

262

Mechanical Technician.  

ERIC Educational Resources Information Center

This document contains 33 units to consider for use in a tech prep competency profile for the occupation of mechanical technician. All the units listed will not necessarily apply to every situation or tech prep consortium, nor will all the competencies within each unit be appropriate. Several units appear within each specific occupation and would…

Ohio State Univ., Columbus. Center on Education and Training for Employment.

263

Cratering mechanics  

NASA Technical Reports Server (NTRS)

Main concepts and theoretical models which are used for studying the mechanics of cratering are discussed. Numerical two-dimensional calculations are made of explosions near a surface and high-speed impact. Models are given for the motion of a medium during cratering. Data from laboratory modeling are given. The effect of gravitational force and scales of cratering phenomena is analyzed.

Ivanov, B. A.

1986-01-01

264

Fluid Mechanics.  

ERIC Educational Resources Information Center

Outlines the contents of Volume II of "Principia" by Sir Isaac Newton. Reviews the contributions of subsequent scientists to the physics of fluid dynamics. Discusses the treatment of fluid mechanics in physics curricula. Highlights a few of the problems of modern research in fluid dynamics. Shows that problems still remain. (CW)

Drazin, Philip

1987-01-01

265

Mural propagation of descending vasa recta responses to mechanical stimulation.  

PubMed

To investigate the responses of descending vasa recta (DVR) to deformation of the abluminal surface, we devised an automated method that controls duration and frequency of stimulation by utilizing a stream of buffer from a micropipette. During stimulation at one end of the vessel, fluorescent responses from fluo4 or bis[1,3-dibutylbarbituric acid-(5)] trimethineoxonol [DiBAC?(3)], indicating cytoplasmic calcium ([Ca²?]CYT) or membrane potential, respectively, were recorded from distant cells. Alternately, membrane potential was recorded from DVR pericytes by nystatin whole cell patch-clamp. Mechanical stimulation elicited reversible [Ca²?)]CYT responses that increased with frequency. Individual pericyte responses along the vessel were initiated within a fraction of a second of one another. Those responses were inhibited by gap junction blockade with 18 ?-glycyrrhetinic acid (100 ?M) or phosphoinositide 3 kinase inhibition with 2-morpholin-4-yl-8-phenylchromen-4-one (50 ?M). [Ca²?]CYT responses were blocked by removal of extracellular Ca²? or L-type voltage-gated channel blockade with nifedipine (10 ?M). At concentrations selective for the T-type channel blockade, mibefradil (100 nM) was ineffective. During mechanostimulation, pericytes rapidly depolarized, as documented with either DiBAC4(3) fluorescence or patch-clamp recording. Single stimuli yielded depolarizations of 22.5 ± 2.2 mV while repetitive stimuli at 0.1 Hz depolarized pericytes by 44.2 ± 4.0 mV. We conclude that DVR are mechanosensitive and that rapid transmission of signals along the vessel axis requires participation of gap junctions, L-type Ca²? channels, and pericyte depolarization. PMID:23698119

Zhang, Zhong; Payne, Kristie; Cao, Chunhua; Pallone, Thomas L

2013-08-01

266

Mechanical Weathering  

NSDL National Science Digital Library

This activity was designed to give students an opportunity to realize that all rocks weather mechanically and each specific rock type has its own particular rate of weathering. Students discover that mechanical weathering is the process of breaking down bedrock into smaller fragments by physical as opposed to chemical means and that rock weathering, although it seems to occur slowly in human terms, is an extremely significant part of the rock cycle. They will learn that weathered rock materials are called sediments and are the structural basis for soils and can also be compacted into sedimentary rock. Students will realize that rock weathering rates vary widely depending on mineral content, texture, rock type, and climate and that differential weathering (varying weathering rates for two or more rock types in physical contact with each other) has given rise to some of the world's most breathtaking scenery.

267

Mechanical capacitor  

NASA Technical Reports Server (NTRS)

A new energy storage system (the mechanical capacitor), using a spokeless magnetically levitated composite ring rotor, is described and design formulas for sizing the components are presented. This new system is configured around a permanent magnet (flux biased) suspension which has active servo control in the radial direction and passive control in the axial direction. The storage ring is used as a moving rotor and electronic commutation of the stationary armature coils is proposed. There is no mechanical contact with the rotating spokeless ring; therefore, long life and near zero rundown losses are projected. A 7-kW h system is sized to demonstrate feasibility. A literature review of flywheel energy storage systems is also presented and general formulas are developed for comparing rotor geometries.

Kirk, J. A.; Studer, P. A.; Evans, H. E.

1976-01-01

268

Mechanical Systems  

Microsoft Academic Search

\\u000a In this chapter, several different control applications for mechanical systems are examined. The first system discussed is\\u000a an autobalancing application. A perfectly balanced rotating object (i.e., the center of geometry and center of mass are coincident)\\u000a will usually not undergo any vibration. However, due to the errors associated with geometric dimensions and the nonhomogeneity\\u000a of the raw material, the construction

Warren E. Dixon; Aman Behal; Darren M. Dawson; Siddharth P. Nagarkatti

269

Biofluid Mechanics  

NASA Astrophysics Data System (ADS)

In contrast to the topics discussed in previous chapters, biofluid mechanics is concerned with flows that are influenced by flexible biological surfaces. We distinguish between flows past living bodies in air or in water, such as bird flight or the swimming of fish, and internal flows, such as the closed blood circulation of living beings. In the previous millions of years, evolution has developed crawling, running, swimming, gliding, and flying as methods of motion of living beings, depending on their size and weight.

Oertel, Herbert

270

Impact Mechanics  

NASA Astrophysics Data System (ADS)

Impact mechanics is concerned with the reaction forces that develop during a collision and the dynamic response of structures to these reaction forces. The subject has a wide range of engineering applications, from designing sports equipment to improving the crashworthiness of automobiles. This book develops several different methodologies for analysing collisions between structures. These range from rigid body theory for structures that are stiff and compact, to vibration and wave analyses for flexible structures. The emphasis is on low-speed impact where damage is local to the small region of contact between the colliding bodies. The analytical methods presented give results that are more robust or less sensitive to initial conditions than have been achieved hitherto. As a text, Impact Mechanics builds upon foundation courses in dynamics and strength of materials. It includes numerous industrially relevant examples and end-of-chapter homework problems drawn from industry and sports. Practising engineers will also find the methods presented in this book useful in calculating the response of a mechanical system to impact.

Stronge, W. J.

2004-03-01

271

The mechanism of mechanical alloying  

Microsoft Academic Search

The mechanical alloying process is a new method for producing composite metal powders with controlled microstructures. It\\u000a is unique in that it is an entirely solid state process, permitting dispersion of insoluble phases such as refractory oxides\\u000a and addition of reactive alloying elements such as aluminum and titanium. Interdispersion of the ingredients occurs by repeated\\u000a cold welding and fracture of

J. S. Benjamin; T. E. Volin

1974-01-01

272

Amphiregulin and PTEN evoke a multimodal mechanism of acquired resistance to PI3K inhibition  

PubMed Central

Phosphoinositide-3 kinase (PI3K) signaling pathway alterations occur broadly in cancer and PI3K is a promising therapeutic target. Here, we investigated acquired resistance to GDC-0941, a PI3K inhibitor in clinical trials. Colorectal cancer (CRC) cells made to be resistant to GDC-0941 were discovered to secrete amphiregulin, which resulted in increased EGFR/MAPK signaling. Moreover, prolonged PI3K pathway inhibition in cultured cells over a period of months led to a secondary loss of PTEN in 40% of the CRC lines with acquired resistance to PI3K inhibition. In the absence of PI3K inhibitor, these PTEN-null PI3K inhibitor-resistant clones had elevated PI3K pathway signaling and decreased sensitivity to MAPK pathway inhibitors. Importantly, PTEN loss was not able to induce resistance to PI3K inhibitors in the absence of amphiregulin, indicating a multimodal mechanism of acquired resistance. The combination of PI3K and MAPK pathway inhibitors overcame acquired resistance in vitro and in vivo. PMID:25053989

Edgar, Kyle A.; Crocker, Lisa; Cheng, Eric; Wagle, Marie-Claire; Wongchenko, Matthew; Yan, Yibing; Wilson, Timothy R.; Dompe, Nicholas; Neve, Richard M.; Belvin, Marcia; Sampath, Deepak; Friedman, Lori S.; Wallin, Jeffrey J.

2014-01-01

273

quantum mechanics  

PubMed Central

-symmetric quantum mechanics (PTQM) has become a hot area of research and investigation. Since its beginnings in 1998, there have been over 1000 published papers and more than 15 international conferences entirely devoted to this research topic. Originally, PTQM was studied at a highly mathematical level and the techniques of complex variables, asymptotics, differential equations and perturbation theory were used to understand the subtleties associated with the analytic continuation of eigenvalue problems. However, as experiments on -symmetric physical systems have been performed, a simple and beautiful physical picture has emerged, and a -symmetric system can be understood as one that has a balanced loss and gain. Furthermore, the phase transition can now be understood intuitively without resorting to sophisticated mathe- matics. Research on PTQM is following two different paths: at a fundamental level, physicists are attempting to understand the underlying mathematical structure of these theories with the long-range objective of applying the techniques of PTQM to understanding some of the outstanding problems in physics today, such as the nature of the Higgs particle, the properties of dark matter, the matter–antimatter asymmetry in the universe, neutrino oscillations and the cosmological constant; at an applied level, new kinds of -synthetic materials are being developed, and the phase transition is being observed in many physical contexts, such as lasers, optical wave guides, microwave cavities, superconducting wires and electronic circuits. The purpose of this Theme Issue is to acquaint the reader with the latest developments in PTQM. The articles in this volume are written in the style of mini-reviews and address diverse areas of the emerging and exciting new area of -symmetric quantum mechanics. PMID:23509390

Bender, Carl M; DeKieviet, Maarten; Klevansky, S. P.

2013-01-01

274

Anticipating mechanisms of resistance to PI3K inhibition in breast cancer: a challenge in the era of precision medicine.  

PubMed

Frequent subversion of the PI3K (phosphoinositide 3-kinase) pathway during neoplastic transformation contributes to several hallmarks of cancer that result in a competitive advantage for cancer cells. Deregulation of this pathway can be the result of genomic alterations such as PIK3CA mutation, PTEN (phosphatase and tensin homologue deleted on chromosome 10) loss or the activation of upstream protein tyrosine kinases. Not surprisingly, the PI3K signalling pathway has become an attractive therapeutic target, and numerous inhibitors are in clinical trials. Unfortunately, current therapies for advanced cancers that target PI3K often lead to the development of resistance and relapse of the disease. It is therefore important to establish the molecular mechanisms of resistance to PI3K-targeted therapy. With the focus on breast cancer, in the present article, we summarize the different ways of targeting PI3K, review potential mechanisms of resistance to PI3K inhibition and discuss the rationale of combination treatments to reach a balance between efficacy and toxicity. PMID:25109950

Leroy, Cedric; Amante, Romain J; Bentires-Alj, Mohamed

2014-08-01

275

Department of Mechanical Engineering-  

E-print Network

Department of Mechanical Engineering- Engineering Mechanics Presidential Council of Alumnae Click #12;The Department of Mechanical Engineering ­ Engineering Mechanics honors its outstanding women Bachelor's of Science Degree in Mechanical Engineering in 1978. She earned her Master's of Science Degree

Endres. William J.

276

Gelation Mechanisms  

NASA Astrophysics Data System (ADS)

In this paper, we survey the gelation mechanisms for various polymeric systems which are classified by the type and the strength of the cross-linkages. These are the "irreversible" gels that are cross-linked chemically by covalent bonds and the "reversible" gels that are cross-linked physically by hydrogen or ionic bonds and by the physical entanglement of polymer chains. Some of the natural polymer gels fall into the class of physical gels, among which the red algae that has attracted attention for various applications is discussed in detail. Various composite gels, formed from mixture of physical and chemical gels are also discussed in the last section of the article. Theoretical models describe the gelation as a process of random linking of subunits to larger and larger molecules by formation of an infinite network, where no matter what type of objects are linked, there is always a critical "gel point" at which the system behaves neither as a liquid nor as a solid on any length scale. The Flory-Stockmayer theory and percolation theory provide bases for modeling this sol-gel phase transition. The experimental techniques for measuring the critical exponents for sol-gel phase transitions in different polymeric systems are introduced and the validation of various theoretical predictions are surveyed.

Pekcan, Önder; Kara, Selim

2012-10-01

277

Umbilical mechanism  

NASA Technical Reports Server (NTRS)

Apparatus is described for automatically mating a pair of connectors and protecting them prior to mating, which minimizes weight and uses relatively simple and reliable mechanisms. Lower and upper connectors (24, 26) are held in lower and upper parts (14, 16) of a housing, with the upper connector mounted on a carrier (32) that is motor driven to move down and mate the connectors. A pair of movable members (36, 38) serve as shields, as coarse alignment aids, and as force transmitters. The movable members are pivotally mounted at the bottom of the upper housing, and as the carrier moves down it pivots the members out of the way. The movable members have socket elements (116) that closely receive pin elements (120) on the lower housing part, to coarsely align the connectors and to react mating and unmating forces between the housings. The carrier has a pair of plate portions (60, 62) with slots (64), and the movable members have cam followers engaged with the slot walls, to move the members with precision. The carrier plate-like portions engage follower members (82) that pivot open lower shield parts (44, 46) covering the lower connector, which is mounted on four stacks of Belleville washers (142).

Barron, Daniel R. (Inventor); Jasulaitis, Vytas (Inventor); Morrill, Brion F. (Inventor)

1995-01-01

278

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 5 problems LAST NAME FIRST NAME #12 with the effective electron mass at the band edges. #12;Applied quantum mechanics 3 (c) Write a computer program

Levi, Anthony F. J.

279

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 1 problems LAST NAME FIRST NAME #12 happens to the beat frequency if the airplane moves in an arc? #12;Applied quantum mechanics 3 Problem 1

Levi, Anthony F. J.

280

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 8 problems LAST NAME FIRST NAME #12;Applied quantum mechanics 3 (b) If the electron is in a semiconductor and has an effective mass m * 0.07 m

Levi, Anthony F. J.

281

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 10 problems LAST NAME FIRST NAME #12 ­( ) L/( )= L/ #12;Applied quantum mechanics 3 (d) Use the results of (b) an (c) to draw the electron

Levi, Anthony F. J.

282

MECHANICAL ENGINEERING The Department of Mechanical Engineer-  

E-print Network

, Solid Mechanics, and Thermal Sciences. Departmental brochures that provide a more detailed description machine, I-DEAS, NX, and AutoCAD. Solid Mechanics The mechanical behavior of advanced materials and applied to solid mechanics problems such as fracture, wave propagation, metal forming, vibration

Ge, Qiaode Jeff

283

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 6 problems LAST NAME FIRST NAME #12 --- and that for a Poisson distribution of such photons #12; 1 2 n ---------------- Applied quantum mechanics 3 (c) Apply conditions is the quantum mechanical result m t 2 2 d d x xd d V x ­= the same Newton's second law in which

Levi, Anthony F. J.

284

MECHANICAL ENGINEERING Ross Schlueter  

E-print Network

MECHANICAL ENGINEERING Ross Schlueter Engineering Deputy For Mechanical Engineering Russ Wells Mechanical Engineering Department Deputy ELECTRONICS, SOFTWARE & INSTRUMENTATION ENGINEERING Henrik von Der Sen Mechanical Admin. Assist. Joan Wolter Electronics Admin. Assist. Marilyn Wong Division Admin

285

Mechanical Engineer Company Description  

E-print Network

Mechanical Engineer Company Description Control Solutions Inc. is a small, dynamic, and rapidly. Position Description The Mechanical Engineer is responsible for all aspects associated with the mechanical enclosures, brackets, cabling assemblies among others. Systems include mechanisms, sensors, hydraulics, among

Kostic, Milivoje M.

286

DEPARTMENT OF MECHANICAL ENGINEERING  

E-print Network

DEPARTMENT OF MECHANICAL ENGINEERING ENGINEERING MECHANICS Research Brochure Message from Message from the Department Chair Dr. William W. Predebon Department of Mechanical Engineering engineering programs in the U.S. In particular, our ABET accredited Mechanical Engineering degree program has

Endres. William J.

287

S-Adenosylmethionine and Methylthioadenosine Inhibit ?-Catenin Signaling by Multiple Mechanisms in Liver and Colon Cancer.  

PubMed

S-Adenosylmethionine (SAMe), the principal methyl donor that is available as a nutritional supplement, and its metabolite methylthioadenosine (MTA) exert chemopreventive properties against liver and colon cancer in experimental models. Both agents reduced ?-catenin expression on immunohistochemistry in a murine colitis-associated colon cancer model. In this study, we examined the molecular mechanisms involved. SAMe or MTA treatment in the colitis-associated cancer model lowered total ?-catenin protein levels by 47 and 78%, respectively. In an orthotopic liver cancer model, increasing SAMe levels by overexpressing methionine adenosyltransferase 1A also reduced total ?-catenin levels by 68%. In both cases, lower cyclin D1 and c-Myc expression correlated with lower ?-catenin levels. In liver (HepG2) and colon (SW480, HCT116) cancer cells with constitutively active ?-catenin signaling, SAMe and MTA treatment inhibited ?-catenin activity by excluding it from the nuclear compartment. However, in liver (Huh-7) and colon (RKO) cancer cells expressing wild-type Wnt/?-catenin, SAMe and MTA accelerated ?-catenin degradation by a glycogen synthase kinase 3-?-dependent mechanism. Both agents lowered protein kinase B activity, but this was not mediated by inhibiting phosphoinositide 3-kinase. Instead, both agents increased the activity of protein phosphatase 2A, which inactivates protein kinase B. The effect of MTA on lowering ?-catenin is direct and not mediated by its conversion to SAMe, as blocking this conversion had no influence. In conclusion, SAMe and MTA inhibit Wnt/?-catenin signaling in colon and liver cancer cells regardless of whether this pathway is aberrantly induced, making them ideal candidates for chemoprevention and/or chemotherapy in these cancers. PMID:25338671

Li, Tony W H; Peng, Hui; Yang, Heping; Kurniawidjaja, Steven; Panthaki, Parizad; Zheng, Yuhua; Mato, José M; Lu, Shelly C

2015-01-01

288

The Department of Mechanical Engineering --Engineering Mechanics  

E-print Network

EYUI? The Department of Mechanical Engineering -- Engineering Mechanics Proudly Presents Professor innovations. In addition, I will discuss cross-validation of fuel- cell measurements from dc 1-V sweep and ac

Endres. William J.

289

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 6 problems LAST NAME FIRST NAME #12 of the system. (b) Find . (c) Find and show that . Under what conditions is the quantum mechanical result( ) td d A t( ) t A td d A /= A B i 2 --- A^ B^,[ ] A^ B^ Et 2 --- n n 1 2 --- #12;Applied quantum

Levi, Anthony F. J.

290

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 9 problems LAST NAME FIRST NAME #12 10 cm 1­ = 2 1� 0.2� µm 3 2 µm 100 µA #12;Applied quantum mechanics 3 Problem 9.4 Modify the computer

Levi, Anthony F. J.

291

Mechanical & Biomedical Engineering  

E-print Network

Mechanical & Biomedical Engineering Department BACHELOR OF SCIENCE IN MECHANICAL ENGINEERING COURSE 105 Mechanical Engineering Graphics 3 CHEM 111L College Chemistry Lab (DLN) 1 ENGL 102 English PHYS 211 Mechanics, Waves & Heat (DLN) 4 UF 100 Intellectual Foundations 3 PHYS 211L Mechanics, Waves

Barrash, Warren

292

Mechanical & Industrial Engineering  

E-print Network

Mechanical & Industrial Engineering 1 Welcome MIE Industrial Advisory Board October 15, 2010 #12;Mechanical & Industrial Engineering 2 MIE Dorothy Adams Undergraduate/Graduate Secretary David Schmidt Associate Professor & Graduate Program Director #12;Mechanical & Industrial Engineering 3 MIE James Rinderle

Mountziaris, T. J.

293

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Katia Bertoldi Harvard University Soft materials in response to diverse stimuli. While the mechanical attributes - such as energy absorption, stiffness and switchable functionalities. Katia Bertoldi is an Assistant Professor of Applied Mechanics at Harvard

294

Mechanical Engineering Graduate Studies  

E-print Network

Mechanical Engineering Graduate Studies Carnegie Mellon University College of Engineering #12;Welcome to Mechanical Engineering! Thank you for your interest in graduate study in the Department of Mechanical Engineering at Carnegie Mellon University. We offer students an excellent educational experience

McGaughey, Alan

295

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Robert J. Hannemann The Gordon Institute and the Department of Mechanical Engineering Tufts University Retooling Our Energy Ecosystem: challenges and Chair of the Tufts Department of Mechanical Engineering. His technical and academic interests

296

Department of Mechanical & Aerospace  

E-print Network

program centers around three general tracks: controls and dynamics, solid mechanics and thermofluids research in the general areas of solid mechanics, dynamics and controls, and thermofluids, with emphases

Acton, Scott

297

Phospholipase A{sub 2} is involved in the mechanism of activation of neutrophils by polychlorinated biphenyls  

SciTech Connect

Aroclor 1242, a mixture of polychlorinated biphenyls (PCBs), activates neutrophils to produce superoxide anion (O{sub 2}{sup {minus}}) by a mechanism that involves phospholipase C-dependent hydrolysis of membrane phosphoinositides; however, subsequent signal transduction mechanisms are unknown. This study determines whether phospholipase A{sub 2}-dependent release of arachidonic acid is involved in PCB-induced O{sub 2}{sup {minus}} production. O{sub 2}{sup {minus}} production was measured in vitro in glycogen-elicited, rat neutrophils in the presence and absence of the inhibitors of phospholipase A{sub 2}: quinacrine, 4-bromophenacyl bromide (BPB), and manoalide. All three agents significantly decreased the amount of O{sub 2}{sup {minus}} detected during stimulation of neutrophils with Aroclor 1242. Similar inhibition occurred when neutrophils were activated with the classical stimuli, formyl-methionyl-leucyl-phenylalanine (fMLP) or phorbol myristate acetate. The effects of BPB and manoalide were not a result of cytotoxicity or other nonspecific effects. Significant release of {sup 3}H-arachidonic acid preceded O{sub 2}{sup {minus}} production in neutrophils stimulated with Aroclor 1242 or fMLP. Manoalide, at a concentration that abolished O{sub 2}{sup {minus}} production, also inhibited the release of {sup 3}H-arachidonate. Aspirin, zileuton, or WEB 2086 did not affect Aroclor 1242-induced O{sub 2}{sup {minus}} production, suggesting that eicosanoids and platelet-activating factor are not needed for neutrophil activation by PCBs. Activation of phos-pholipase A{sub 2} and O{sub 2}{sup {minus}} production do not appear to involve the Ah receptor. These data suggest that Aroclor 1242 stimulates neutrophils to produce O{sub 2}{sup {minus}} by a mechanism that involves phospholipase A{sub 2}-dependent release of arachiodonic acid. 49 refs., 6 figs., 2 tabs.

Tithof, P.K.; Schiamberg, E.; Ganey, P.E. [Univ. of Michigan, Ann Arbor, MI (United States); Peters-Golden, M. [Michigan State Univ., East Lansing, MI (United States)

1996-01-01

298

Applied quantum mechanics 1 Applied Quantum Mechanics  

E-print Network

Applied quantum mechanics 1 Applied Quantum Mechanics Chapter 5 problems LAST NAME FIRST NAME #12 + --------------------------------------------- k = t 10/= t 1­= Ek 2t kxL( ) 2t 2kxL( )cos+cos= t 10/= t 1­= t 0.2­= #12;Applied quantum mechanics 3 (c) Write a computer program to plot the electron density of states for a square lat- tice

Levi, Anthony F. J.

299

Cell mechanical modeling and mechanical properties characterization.  

E-print Network

???Cell biomechanics regulate cellular physiological functions, such as locomotion, cell division, proliferation, mechanotransduction, and cell death. Current research reports that alterations of the mechanical properties… (more)

Tan, Youhua (???)

2010-01-01

300

Signaling mechanisms mediating muscarinic enhancement of GABAergic synaptic transmission in the spinal cord.  

PubMed

Activation of muscarinic acetylcholine receptors (mAChRs) inhibits spinal nociceptive transmission by potentiation of GABAergic tone through M(2), M(3), and M(4) subtypes. To study the signaling mechanisms involved in this unique mAChR action, GABAergic spontaneous inhibitory postsynaptic currents (sIPSCs) of lamina II neurons were recorded using whole-cell patch clamp techniques in rat spinal cord slices. The mAChR agonist oxotremorine-M caused a profound increase in the frequency of GABAergic sIPSCs, which was abolished in the Ca(2+)-free solution. Inhibition of voltage-gated Ca(2+) channels with Cd(2+) and Ni(2+) largely reduced the effect of oxotremorine-M on sIPSCs. Blocking nonselective cation channels (NSCCs) with SKF96365 or 2-APB also largely attenuated the effect of oxotremorine-M. However, the KCNQ channel blocker XE991 and the adenylyl cyclase inhibitor MDL12330A had no significant effect on oxotremorine-M-induced increases in sIPSCs. Furthermore, the phosphoinositide-3-kinase (PI3K) inhibitor wortmannin or LY294002 significantly reduced the potentiating effect of oxotremorine-M on sIPSCs. In the spinal cord in which the M(3) subtype was specifically knocked down by intrathecal small interfering RNA (siRNA) treatment, SKF96365 and wortmannin still significantly attenuated the effect of oxotremorine-M. In contrast, SKF96365 and wortmannin both failed to alter the effect of oxotremorine-M on sIPSCs when the M(2)/M(4) mAChRs were blocked. Therefore, our study provides new evidence that activation of mAChRs increases synaptic GABA release through Ca(2+) influx and voltage-gated Ca(2+) channels. The PI3K-NSCC signaling cascade is primarily involved in the excitation of GABAergic interneurons by the M(2)/M(4) mAChRs in the spinal dorsal horn. PMID:19110040

Zhang, H-M; Chen, S-R; Cai, Y-Q; Richardson, T E; Driver, L C; Lopez-Berestein, G; Pan, H-L

2009-02-18

301

Mechanical & Industrial Engineering  

E-print Network

Mechanical & Industrial Engineering 1 Welcome MIE Industrial Advisory Board May 5th, 2011 #12;Mechanical & Industrial Engineering 2 IAB 2010-2011 · David K. Anderson ­ Alden Research Laboratory, Inc went on for three weeks Mechanical & Industrial Engineering 6 #12;Reza Shahbazian Yassar Mechanical

Mountziaris, T. J.

302

Defense Mechanisms: A Bibliography.  

ERIC Educational Resources Information Center

This bibliography includes studies of defense mechanisms, in general, and studies of multiple mechanisms. Defense mechanisms, briefly and simply defined, are the unconscious ego defendants against unpleasure, threat, or anxiety. Sigmund Freud deserves the clinical credit for studying many mechanisms and introducing them in professional literature.…

Pedrini, D. T.; Pedrini, Bonnie C.

303

Mechanical Engineering Graduate Student  

E-print Network

Mechanical Engineering Graduate Student Handbook January 2014 Department of Mechanical Engineering University of Wisconsin-Madison #12;Mechanical Engineering Web Page: http://www.engr.wisc.edu/me Graduate & Terrace Chairs) and Samantha Stepp (ME Building) 1 #12;Department of Mechanical Engineering AGUIDE

Wisconsin at Madison, University of

304

Mechanical Engineering & Thermal Group  

E-print Network

Mechanical Engineering & Thermal Group The Mechanical Engineering (ME) & Thermal Group at LASP has, and ground- based mechanical systems. Instrument Design Building on decades of design experience that has evolved with the complexity of instrument design demands, LASP mechanical engineers develop advanced

Mojzsis, Stephen J.

305

Mechanical Engineering Undergraduate  

E-print Network

Mechanical Engineering Department Undergraduate Advising Manual for Bachelor of Science Degrees in Mechanical Engineering and Engineering Mechanics 2011-2012 - Updated April 15, 2012 #12;Johns Hopkins University ­ Department of Mechanical Engineering 2011-2012 Undergraduate Student Advising Manual Page 2

Ghosh, Somnath

306

Mechanical Engineering Undergraduate  

E-print Network

Mechanical Engineering Department Undergraduate Advising Manual for Bachelor of Science Degrees in Mechanical Engineering and Engineering Mechanics 2012-2013 - Updated July 14, 2013 #12;Johns Hopkins University ­ Department of Mechanical Engineering 2012-2013 Undergraduate Student Advising Manual Page 2

Ghosh, Somnath

307

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar James Bird Department of Mechanical Engineering Boston ­ are discussed. James Bird is an Assistant Professor in the Mechanical Engineering Department at Boston completed post-doctoral research at MIT. His research interests include experimental fluid mechanics

308

Mechanics rules cell biology  

Microsoft Academic Search

Cells in the musculoskeletal system are subjected to various mechanical forces in vivo. Years of research have shown that these mechanical forces, including tension and compression, greatly influence various cellular functions such as gene expression, cell proliferation and differentiation, and secretion of matrix proteins. Cells also use mechanotransduction mechanisms to convert mechanical signals into a cascade of cellular and molecular

James HC Wang; Bin Li

2010-01-01

309

INTRODUCTION TO THE MECHANICS  

E-print Network

INTRODUCTION TO THE MECHANICS OF A CONTINUOUS MEDIUM Lawrence E. Malvern Professor of Mechanics princi- ples common to all branches of solid and fluid mechanics, designed to appeal to the intuition science. The book arose from the need to provide a general preparation in contin- uum mechanics

Kaminski, Edouard

310

Mechanical Engineering Course  

Microsoft Academic Search

Carnegie Mellon University offers a first-year course titled Funda- mentals of Mechanical Engineering to introduce undergraduate students to the discipline of mechanical engineering. The goals of the course are to excite students about the field of mechanical engi- neering early in their careers, introduce basic mechanical engineer- ing concepts in an integrated way, provide a link to the basic physics

SUSAN A. AMBROSE; CRISTINA H. AMON

311

Quantum Mechanics + Open Systems  

E-print Network

Quantum Mechanics + Open Systems = Thermodynamics ? Jochen Gemmer T¨ubingen, 09.02.2006 #12., World Scientific) #12;Fundamental Law or Emergent Description? Quantum Mechanics i t = (- 2 2m + V or Emergent Description? Quantum Mechanics i t = (- 2 2m + V ) "Heisenberg Cut" Classical Mechanics: m d2

Steinhoff, Heinz-Jürgen

312

Engineering. Mechanical Engineering.  

E-print Network

Department Of Mechanical Engineering. Studying Mechanical Engineering. www.sheffield.ac.uk/mecheng #12;from the Head of Department A degree in mechanical engineering is the key to an exciting course is an MEng or BEng in Mechanical Engineering. You can also learn a language and study abroad

Stevenson, Mark

313

graduate programs mechanical engineering  

E-print Network

graduate programs mechanical engineering engineering.asu.edu/graduate/mech Arizona State University offers the following graduate degrees in mechanical engineering: GRADUATE DEGREE PROGRAMS Master of Science ­ Mechanical Engineering (Thesis; M.S.) Master of Science in Engineering ­ Mechanical Engineering

Zhang, Junshan

314

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Yoed Rabin Department of Mechanical Engineering Carnegie at Carnegie Mellon University. Yoed Rabin is a Professor of Mechanical Engineering at Carnegie Mellon University. He received his DSc in Mechanical Engineering in 1994 from the Technion ­ Israel Institute

315

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Virginia Ferguson Department of Mechanical Engineering engineering of this interface. Virginia Ferguson (Ph.D. Mechanical Engineering, 2001) is Assistant Professor of Mechanical Engineering at the Univer- sity of Colorado, Boulder since 2006. She held a joint postdoctoral

316

ENGINEERING MECHANICS SEMINARSENGINEERING MECHANICS SEMINARS THINK COMPOSITE  

E-print Network

. The first part presents the concept of nano-composite interlayers introduced/integrated into the traditional(St-co-GMA) and P(St-co-GMA)/MWCNT electrospun nano-fibers are applied on conventional carbon/epoxy prepreg pliesENGINEERING MECHANICS SEMINARSENGINEERING MECHANICS SEMINARS THINK COMPOSITE "THINK outside the box

Ponce, V. Miguel

317

Minocycline ameliorates LPS-induced inflammation in human monocytes by novel mechanisms including LOX-1, Nur77 and LITAF inhibition  

PubMed Central

Background Minocycline exhibits anti-inflammatory properties independent of its antibiotic activity, ameliorating inflammatory responses in monocytes and macrophages. However, the mechanisms of minocycline anti-inflammatory effects are only partially understood. Methods Human circulating monocytes were cultured in the presence of lipopolysaccharide (LPS), 50 ng/ml, and minocycline (10–40 µM). Gene expression was determined by RT-PCR, cytokine and prostaglandin E2 (PGE2) release by ELISA, protein expression, phosphorylation and nuclear translocation by Western blotting. Results Minocycline significantly reduced the inflammatory response in LPS-challenged monocytes, decreasing LPS-induced transcription of pro-inflammatory tumor-necrosis factor alpha (TNF-?), interleukin-1 beta, interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2), and the LPS-stimulated TNF-?, IL-6 and PGE2 release. Minocycline inhibited LPS-induced activation of the lectin-like oxidized low density lipoprotein receptor-1 (LOX-1), NF-?B, LPS-induced TNF-? factor (LITAF) and the Nur77 nuclear receptor. Mechanisms involved in the anti-inflammatory effects of minocycline include a reduction of LPS-stimulated p38 mitogen-activated protein kinase (p38 MAPK) activation and stimulation of the phosphoinositide 3-kinase (PI3K)/Akt pathway. Conclusions We provide novel evidence demonstrating that the anti-inflammatory effects of minocycline in human monocytes include, in addition to decreased NF-?B activation, abrogation of the LPS-stimulated LOX-1, LITAF, Nur77 pathways, p38 MAPK inhibition and PI3K/Akt activation. Our results reveal that minocycline inhibits points of convergence of distinct and interacting signaling pathways mediating multiple inflammatory signals which may influence monocyte activation, traffic and recruitment into the brain. General significance Our results in primary human monocytes contribute to explain the profound anti-inflammatory and protective effects of minocycline in cardiovascular and neurological diseases and may have direct translational relevance. PMID:22306153

Pang, Tao; Wang, Juan; Benicky, Julius; Saavedra, Juan M.

2012-01-01

318

Inhibition of Ca2+-independent phospholipase A2 decreases prostate cancer cell growth by p53-dependent and independent mechanisms.  

PubMed

The mechanisms by which Ca(2+)-independent phospholipase A(2) (iPLA(2)) mediates cell growth in p53-positive LNCaP and p53-negative PC-3 prostate cancer cell lines were studied. Exposure of cells to the iPLA(2) selective inhibitor bromoenol lactone (BEL; 0-20 microM) induced concentration- and time-dependent decreases in cell growth based on 3-(4, dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide staining and cell number. Decreased cell growth was not caused by cell death as BEL exposure did not alter nuclear morphology or increase annexin V (apoptotic cell marker) or propidium iodide (necrotic cell marker) staining after 48 h. Decreased growth correlated to a G(1)/G(0) arrest in LNCaP cells and aG(2)/M arrest in PC-3 cells. In LNCaP cells, G(1) arrest was preceded by time- (0-48 h) and concentration-dependent (0-10 microM) increases in the expression of the tumor suppressor protein p53 and the cyclin-dependent kinase inhibitor p21. Increases in p53 expression preceded increases in p21 expression by 8 h. In LNCaP cells, BEL treatment decreased the expression of the p53 antagonist Mdm2, while increasing Akt phosphorylation. BEL treatment also increased Akt phosphorylation in PC-3 cells, but Mdm2 was not detected. The ability of BEL to increase Akt phosphorylation was inhibited by the phosphoinositide 3-kinase inhibitor LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one]. BEL treatment also decreased agonist-induced activation of the epidermal growth factor receptor. These data suggest that inhibition of iPLA(2) decreases prostate cancer cell growth by p53-dependent and independent mechanisms. Furthermore, alterations in Mdm2 and epidermal growth factor receptor activation following BEL exposure suggest novel roles for iPLA(2) in prostate cancer cell signaling. PMID:18441250

Sun, Bin; Zhang, Xiaoling; Talathi, Sonia; Cummings, Brian S

2008-07-01

319

Central neural mechanisms governing postural cardiovascular mechanisms  

NASA Technical Reports Server (NTRS)

The results of the vestibular apparatus and cerebellum in orthostatic reflex control are summarized. Mechanisms within the brain which govern circulation reflexes and the consequences of disturbances in their function are also included.

Reis, D. J.

1976-01-01

320

External Resource: Mechanical Weathering  

NSDL National Science Digital Library

A student activity with teacher's sheet, to give the students an opportunity to realize that all rocks weather mechanically and each specific rock type has its own particular rate of weathering. Mechanical weathering is the process of breaking down bedroc

1900-01-01

321

Astrophysics QuantumMechanics  

E-print Network

Astrophysics Geometry QuantumMechanics Stochasticanalysis DifferentialEquations A N N U A L R E P O report 2010 6 Geometry 6 Stochastic analysis 8 Differential Equations 9 Astrophysics 11 Quantum Mechanics

Johansen, Tom Henning

322

STUDENT HANDBOOK MECHANICAL ENGINEERING  

E-print Network

2012-2013 STUDENT HANDBOOK MECHANICAL ENGINEERING AEROSPACE ENGINEERING ENVIRONMENTAL ENGINEERING accredited programs) Aerospace and Mechanical Engineering: · An ability to apply knowledge of mathematics to have successful careers as engineers, technology leaders and innovators. To prepare our students

Krstic, Miroslav

323

Programmable Mechanical Metamaterials  

E-print Network

We create mechanical metamaterials whose response to uniaxial compression can be programmed by lateral confinement, allowing monotonic, non-monotonic and hysteretic behavior. These functionalities arise from a broken rotational symmetry which causes highly nonlinear coupling of deformations along the two primary axes of these metamaterials. We introduce a soft mechanism model which captures the programmable mechanics, and outline a general design strategy for confined mechanical metamaterials. Finally, we show how inhomogeneous confinement can be explored to create multi stability and giant hysteresis.

Bastiaan Florijn; Corentin Coulais; Martin van Hecke

2014-07-16

324

Mechanical & Aerospace Engineering  

E-print Network

/Dissertation................................................................................................ 36 Classroom Protocol ........................................................................................................... 44 Departmental Safety Regulations, Manufacture & Engineering Management; Electronic & Electrical Engineering; Mechanical & Aerospace Engineering

Strathclyde, University of

325

Mechanical & Aerospace Engineering  

E-print Network

/Dissertation................................................................................................ 35 Classroom Protocol ........................................................................................................... 43 Departmental Safety Regulations, Manufacture & Engineering Management; Electronic & Electrical Engineering; Mechanical & Aerospace Engineering

Strathclyde, University of

326

Programmable Mechanical Metamaterials  

NASA Astrophysics Data System (ADS)

We create mechanical metamaterials whose response to uniaxial compression can be programmed by lateral confinement, allowing monotonic, nonmonotonic, and hysteretic behavior. These functionalities arise from a broken rotational symmetry which causes highly nonlinear coupling of deformations along the two primary axes of these metamaterials. We introduce a soft mechanism model which captures the programmable mechanics, and outline a general design strategy for confined mechanical metamaterials. Finally, we show how inhomogeneous confinement can be explored to create multistability and giant hysteresis.

Florijn, Bastiaan; Coulais, Corentin; van Hecke, Martin

2014-10-01

327

Programmable mechanical metamaterials.  

PubMed

We create mechanical metamaterials whose response to uniaxial compression can be programmed by lateral confinement, allowing monotonic, nonmonotonic, and hysteretic behavior. These functionalities arise from a broken rotational symmetry which causes highly nonlinear coupling of deformations along the two primary axes of these metamaterials. We introduce a soft mechanism model which captures the programmable mechanics, and outline a general design strategy for confined mechanical metamaterials. Finally, we show how inhomogeneous confinement can be explored to create multistability and giant hysteresis. PMID:25379923

Florijn, Bastiaan; Coulais, Corentin; van Hecke, Martin

2014-10-24

328

Reputation Mechanisms Elodie Fourquet  

E-print Network

Reputation Mechanisms Elodie Fourquet efourque@cgl.uwaterloo.ca University of Waterloo Abstract to be created. The purpose of a reputation mechanism is to maintain trust among participants in order to produce more social welfare. We analyse why current reputation mechanisms are and are not successful

Waterloo, University of

329

Mechanical & Industrial Engineering  

E-print Network

Mechanical & Industrial Engineering Mario A. Rotea Professor and Department Head #12;2Mechanical & Industrial Engineering Outline · Undergraduate Degree Programs · Graduate Degree Programs · The Faculty · The Research · Summary #12;3Mechanical & Industrial Engineering Undergraduate Programs ­ BSME & BSIE 0 20 40 60

Mountziaris, T. J.

330

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Shuodao Wang Postdoctoral Fellow University of Illinois, Urbana-Champaign Mechanical Design and Fabrication Techniques for Bio-Electronic Systems 11:00 AM Friday that bridge this gap in mechanics and form will create new opportunities in bio-inspired and bio

Lin, Xi

331

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Maureen Lynch Postdoctoral Fellow Cornell University Mechanical Loading Decreases Osteolysis and Tumor Formation via Effects on Bone Remodeling 11:00 AM Friday to mechanical stimuli in the skeleton, yet the role of biomechanical loading remains poorly characterized

Lin, Xi

332

Missing Mechanism Information  

ERIC Educational Resources Information Center

The first recommendation Kazdin made for advancing the psychotherapy research knowledge base, improving patient care, and reducing the gulf between research and practice was to study the mechanisms of therapeutic change. He noted, "The study of mechanisms of change has received the least attention even though understanding mechanisms may well be…

Tryon, Warren W.

2009-01-01

333

Fractional quantum mechanics  

Microsoft Academic Search

A path integral approach to quantum physics has been developed. Fractional path integrals over the paths of the Lévy flights are defined. It is shown that if the fractality of the Brownian trajectories leads to standard quantum and statistical mechanics, then the fractality of the Lévy paths leads to fractional quantum mechanics and fractional statistical mechanics. The fractional quantum and

Nikolai Laskin

2000-01-01

334

Statistical Mechanics its applications  

E-print Network

Statistical Mechanics and its applications Dr A. Alavi Part II Chemistry 2007-2008 1 #12;I. INTRODUCTION Statistical Mechanics is concerned with how to describe the behaviour of systems containing large, and how this is done, as well as its theoretical justification, is the subject of Statistical Mechanics. A

Alavi, Ali

335

Giovanni Gallavotti STATISTICAL MECHANICS  

E-print Network

Giovanni Gallavotti STATISTICAL MECHANICS Short Treatise Roma 1999 #12;. #12;Short treatise of Statistical Mechanics Giovanni Gallavotti Dipartimento di Fisica Universit`a di Roma La Sapienza 00185 Roma that started with my involvement as Coordinator of the Statistical Mechanics section of the Italian Encyclo

Roma "La Sapienza", Università di

336

Giovanni Gallavotti STATISTICAL MECHANICS  

E-print Network

Giovanni Gallavotti STATISTICAL MECHANICS Short Treatise Roma 1999 #12; . #12; Short treatise of Statistical Mechanics Giovanni Gallavotti Dipartimento di Fisica Universitâ??a di Roma La Sapienza 00185 Roma that started with my involvement as Coordinator of the Statistical Mechanics section of the Italian Encyclo

Roma "La Sapienza", Università di

337

Basic Engineer Equipment Mechanic.  

ERIC Educational Resources Information Center

This student guide, one of a series of correspondence training courses designed to improve the job performance of members of the Marine Corps, deals with the skills needed by basic engineer equipment mechanics. Addressed in the four individual units of the course are the following topics: mechanics and their tools (mechanics, hand tools, and power…

Marine Corps Inst., Washington, DC.

338

Introduction to Quantum Mechanics  

E-print Network

The purpose of this contribution is to give a very brief introduction to Quantum Mechanics for an audience of mathematicians. I will follow Segal's approach to Quantum Mechanics paying special attention to algebraic issues. The usual representation of Quantum Mechanics on Hilbert spaces is also discussed.

Eduardo J. S. Villaseñor

2008-04-23

339

Mechanical & Aerospace Engineering  

E-print Network

Mechanical & Aerospace Engineering It is a new beginning for innovative fundamental and applied Stability and Mechanical Properties Mark Tschopp Materials Engineer U.S. Army Research Laboratory September and consolidation of bulk nanocrystalline materials using mechanical alloying, the alloy development and synthesis

340

Department of Mechanical Engineering  

E-print Network

Department of Mechanical Engineering Undergraduate Program 2006-2007 Northern Illinois University DeKalb, IL #12;DEPARTMENT OF MECHANICAL ENGINEERING NORTHERN ILLINOIS UNIVERSITY Web: http://www.ceet.niu.edu/depts/me/ Phone: 815-753-9979 The mission of the Department of Mechanical Engineering is to provide an up

Karonis, Nicholas T.

341

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar William P. King Professor University of Illinois. William P. King is the College of Engineering Bliss Professor in the Department of Mechanical Science and Engineering and Electrical and Computer Engineering. He received the Ph.D. degree in mechanical engineering

342

Department of Mechanical Engineering  

E-print Network

Department of Mechanical Engineering Undergraduate Program 2007-2008 Northern Illinois University DeKalb, IL #12;DEPARTMENT OF MECHANICAL ENGINEERING NORTHERN ILLINOIS UNIVERSITY Web: http://www.ceet.niu.edu/depts/me/ Phone: 815-753-9979 The mission of the Department of Mechanical Engineering is to provide an up

Karonis, Nicholas T.

343

Triple seesaw mechanism  

E-print Network

On fitting the type II seesaw mechanism into the type I seesaw mechanism, we obtain a formula to the neutrino masses which get suppressed by high-scale $M^3$ in its denominator. As a result, light neutrinos are naturally obtained with new physics at TeV scale. As interesting consequence, the mechanism may be directly probed at the LHC by directly producing the TeV states intrinsic of the mechanism. We show that the 3-3-1 model with right-handed neutrinos realizes naturally such seesaw mechanism.

D. Cogollo; H. Diniz; C. A. de S. Pires

2010-02-09

344

Phosphoinositide isoforms determine compartment-specific ion channel activity  

E-print Network

pathways. To determine whether intracellularly localized channels are simi- larly "inactivated" at the PM, we studied PIP2 modulation of in- tracellular TRPML1 channels. TRPML1 channels are primarily in a common PIP2-interacting domain. Thus, PI(4,5)P2 may serve as a negative cofactor for intracellular

Xu, Haoxing

345

Research article Expression and evolution of the phosphoinositide-specific  

E-print Network

, Ames, IA 50011, USA c Department of Plant Pathology, Iowa State University, Ames, IA 50011, USA. Plant PI-PLCs are structurally close to the mammalian PI-PLC-z isoform. The Arabidopsis genome contains environmental stimuli were studied by applying a quantitative RTePCR approach. Multiple members of the gene

Brendel, Volker

346

Phosphoinositides and membrane traffic in health and disease  

Microsoft Academic Search

The phosphorylated derivatives of phosphatidylinositol (PtdIns) are collectively known as the polyphosphoinositides (PIs) and they were originally identified as precursors of second messengers. In particular, PtdIns 4,5-bisphosphate (PtdIns45P2), initially the most studied of the PIs, was shown to be the substrate of a PI-specific phospholipase C (PI-PLC), which upon agonist stimulation generates the water-soluble inositol 1,4,5-trisphosphate and the membrane-bound diacylglycerol.

Anna Godi; Antonella Di Campli; Maria Antonietta De Matteis

347

Cytosolic phospholipase A2 is coupled to muscarinic receptors in the human astrocytoma cell line 1321N1: characterization of the transducing mechanism.  

PubMed Central

The cholinergic agonist carbachol induced the release of arachidonic acid in the 1321N1 astrocytoma cell line, and this was blocked by atropine, suggesting the involvement of muscarinic receptors. To assess the mechanisms of signalling involved in the response to carbachol, a set of compounds characterized by eliciting responses through different mechanisms was tested. A combination of 4beta-phorbol 12beta-myristate 13alpha-acetate and thapsigargin, an inhibitor of endomembrane Ca2+-ATPase that induces a prolonged elevation of cytosolic Ca2+ concentration, induced an optimal response, suggesting at first glance that both protein kinase C (PKC) and Ca2+ mobilization were involved in the response. This was consistent with the observation that carbachol elicited Ca2+ mobilization and PKC-dependent phosphorylation of cytosolic phospholipase A2 (cPLA2; phosphatide sn-2-acylhydrolase, EC 3.1.1.4) as measured by a decrease in electrophoretic mobility. Nevertheless, the release of arachidonate induced by carbachol was unaltered in media containing decreased concentrations of Ca2+ or in the presence of neomycin, a potent inhibitor of phospholipase C which blocks phosphoinositide turnover and Ca2+ mobilization. Guanosine 5'-[gamma-thio]triphosphate added to the cell-free homogenate induced both [3H]arachidonate release and cPLA2 translocation to the cell membrane fraction in the absence of Ca2+, thus suggesting the existence of an alternative mechanism of cPLA2 translocation dependent on G-proteins and independent of Ca2+ mobilization. From the combination of experiments utilizing biochemical and immunological tools the involvement of cPLA2 was ascertained. In summary, these data indicate the existence in the astrocytoma cell line 1321N1 of a pathway involving the cPLA2 which couples the release of arachidonate to the occupancy of receptors for a neurotransmitter, requires PKC activity and G-proteins and might operate in the absence of Ca2+ mobilization. PMID:9173894

Bayon, Y; Hernandez, M; Alonso, A; Nunez, L; Garcia-Sancho, J; Leslie, C; Sanchez Crespo, M; Nieto, M L

1997-01-01

348

MechanicalEngineering The Department of Mechanical Engineering presents  

E-print Network

Colloquium MechanicalEngineering The Department of Mechanical Engineering presents: Dr. Julia R is the development of innovative experimental approaches to as- sess mechanical properties, microstructure evolution

349

Space Mechanisms Technology Workshop  

NASA Technical Reports Server (NTRS)

The Mechanical Components Branch at NASA Glenn Research Center hosted a workshop on Tuesday, May 14, 2002, to discuss space mechanisms technology. The theme for this workshop was 'Working in the Cold,' a focus on space mechanisms that must operate at low temperatures. We define 'cold' as below -60C (210 K), such as would be found near the equator of Mars. However, we are also concerned with much colder temperatures such as in permanently dark craters of the Moon (about 40 K).

Oswald, Fred B. (Editor)

2002-01-01

350

Mechanical devices: A compilation  

NASA Technical Reports Server (NTRS)

A collection of new technology items that should be of interest to mechanical engineers, machinists, and others who design or work with mechanical devices was described. Section 1 contains articles on several new or modified tools, Section 2 describes a number of specialized mechanical systems, and the last section is devoted to valves, bearings, and other parts that might be used with larger systems. The last patent information available is also given.

1976-01-01

351

Positioning Mechanism For Hoisting  

NASA Technical Reports Server (NTRS)

Mechanism positions large, heavy objects in container for lifting out by hoist, crane, or winch. Handles objects gently and ensures they are lifted cleanly away in vertical direction without bumping container. Developed for lifting offset pieces of solid-propellant core out of rocket motor through its propellant port. Similar specialized mechanisms can be developed to lift other specially shaped, specially contained heavy objects. Track in base of mechanism guides each trunnion and piece to which attached to middle as hydraulic rods extend. When mechanism lifted, tilted pieces swing inward and come to rest on energy-absorbing paddle.

Marlin, John D., III; Moore, Barry J.; Myers, Robert I.

1992-01-01

352

More on Atonic Mechanics  

NASA Astrophysics Data System (ADS)

We have shown that crystal based experiments, such as those of Davisson and Germer, do not empirically verify de Broglie's matter-wave hypothesis. We named this theory Atonic Mechanics. This mechanics has also been used to accurately calculate the hundreds of helium atom energy levels tabulated by NIST. We have joined Atonic Mechanics with Einstein's General Relativity. Now we show how fractional values of h-bar arise for the angular momentum of the helium atom in this theory. We now also discuss energy and momentum in Atonic Mechanics.

Phillips, Alfred

2007-03-01

353

Mechanisms for space applications  

NASA Astrophysics Data System (ADS)

All space instruments contain mechanisms or moving mechanical assemblies that must move (sliding, rolling, rotating, or spinning) and their successful operation is usually mission-critical. Generally, mechanisms are not redundant and therefore represent potential single point failure modes. Several space missions have suffered anomalies or failures due to problems in applying space mechanisms technology. Mechanisms require a specific qualification through a dedicated test campaign. This paper covers the design, development, testing, production, and in-flight experience of the PICARD/SODISM mechanisms. PICARD is a space mission dedicated to the study of the Sun. The PICARD Satellite was successfully launched, on June 15, 2010 on a DNEPR launcher from Dombarovskiy Cosmodrome, near Yasny (Russia). SODISM (SOlar Diameter Imager and Surface Mapper) is a 11 cm Ritchey-Chretien imaging telescope, taking solar images at five wavelengths. SODISM uses several mechanisms (a system to unlock the door at the entrance of the instrument, a system to open/closed the door using a stepper motor, two filters wheels using a stepper motor, and a mechanical shutter). For the fine pointing, SODISM uses three piezoelectric devices acting on the primary mirror of the telescope. The success of the mission depends on the robustness of the mechanisms used and their life.

Meftah, M.; Irbah, A.; Le Letty, R.; Barré, M.; Pasquarella, S.; Bokaie, M.; Bataille, A.; Poiet, G.

2012-06-01

354

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Heather Clark Associate Professor Northeastern University of Pharmaceutical Sciences at Northeastern University. Dr. Clark obtained her graduate degree in Chemistry from

Lin, Xi

355

Photon Wave Mechanics  

E-print Network

In contrast to wave functions in nonrelativistic quantum mechanics interpreted as probability amplitudes, wave functions in relativistic quantum mechanics have generalized meanings such as charge-density amplitudes, energy-density amplitudes as well as particle-number density amplitudes, etc. Applying electromagnetic field intensities we construct a photon wave function, it corresponds to the (1,0)+(0,1) spinor representation of the electromagnetic field, and can be interpreted as the energy-density amplitude of photons outside a source. In terms of photon wave functions we develop photon wave mechanics, which provides us with a new quantum-mechanical description for photons outside a source.

Zhi-Yong Wang; Cai-Dong Xiong; Ole Keller

2005-11-18

356

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Nicholas Zabaras Professor Cornell University Uncertainty media requires innovations in mathematical and computational thinking. While multiscale approaches have

357

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Sameer Sonkusale Associate Professor Tufts Univeristy of single chip sensors for chemical sensing. First, an innovative approach termed "CMOS for Nanoas- sembly

Lin, Xi

358

Experimental unsaturated soil mechanics  

Microsoft Academic Search

In this general report, experimental systems and procedures of investigating the hydro-mechanical behaviour of unsaturated soils are presented. The water retention properties of unsaturated soils are commented and linked to various physical parameters and properties of the soils. Techniques of controlling suction are described together with their adaptation in various laboratory testing devices. Some typical features of the mechanical behaviour

Pierre Delage

2008-01-01

359

Safety Critical Mechanisms  

NASA Technical Reports Server (NTRS)

Spaceflight mechanisms have a reputation for being difficult to develop and operate successfully. This reputation is well earned. Many circumstances conspire to make this so: the environments in which the mechanisms are used are extremely severe, there is usually limited or no maintenance opportunity available during operation due to this environment, the environments are difficult to replicate accurately on the ground, the expense of the mechanism development makes it impractical to build and test many units for long periods of time before use, mechanisms tend to be highly specialized and not prone to interchangeability or off-the-shelf use, they can generate and store a lot of energy, and the nature of mechanisms themselves, as a combination of structures, electronics, etc. designed to accomplish specific dynamic performance, makes them very complex and subject to many unpredictable interactions of many types. In addition to their complexities, mechanism are often counted upon to provide critical vehicle functions that can result in catastrophic events should the functions not be performed. It is for this reason that mechanisms are frequently subjected to special scrutiny in safety processes. However, a failure tolerant approach, along with good design and development practices and detailed design reviews, can be developed to allow such notoriously troublesome mechanisms to be utilized confidently in safety-critical applications.

Robertson, Brandan

2008-01-01

360

Mechanics of Composite Materials  

Microsoft Academic Search

The mechanical behavior of composites is traditionally evaluated on both microscopic and macroscopic scale to take into account inhomogeneity. Micromechanics attempts to quantify the interactions of fiber and matrix (reinforcement and resin) on a microscopic scale on par with the diameter of a single fiber. Macromechanics treats composites as homogeneous materials, with mechanical properties representative of the laminate as a

Robert M. Jones

1999-01-01

361

Mechanical Booster Pump  

NSDL National Science Digital Library

This resource, hosted by Maricopa Advanced Technology Education Center (MATEC), is an animation of a mechanical booster (rotary lobe) pump. The objective is to describe the components and operation of a mechanical booster pump (Roots Blower). This simulation is from Module 99 of the vacuum technology and gas control cluster of the MATEC Module Library (MML).

2011-06-27

362

Consider mechanical recompression evaporation  

Microsoft Academic Search

Mechanical recompression is a technique for improving latent heat management by recycling the latent heat of evaporation. The product vapor is isotropically compressed to a higher temperature and pressure so that it can be used as the heating medium that drives the evaporation. The mechanical recompression evaporator typically requires no main process steam, cooling water, or process steam condenser. The

1994-01-01

363

Time in quantum mechanics  

E-print Network

The role of time in quantum mechanics has been and is still very controversial. The purpose of this paper was to explore the historical interpretation of time in quantum mechanics, to determine the current status of this problem-L and to investigate...

Chapin, Kimberly R.

2012-06-07

364

Quantum Mechanics Measurements, Mutually  

E-print Network

Quantum Mechanics Measurements, Mutually Unbiased Bases and Finite Geometry Or why six is the first) #12;Quantum Mechanics for Dummies Finite dimensional quantum states are represented by trace one,1 -icS1,1[ ] #12;Quantum systems evolve and are measured. The evolution of a quantum system using

Gruner, Daniel S.

365

Advanced Visual Quantum Mechanics  

NSDL National Science Digital Library

This page provides links to a range of teaching materials for use in an upper-level undergraduate quantum mechanics course. These are developed from some of the concepts of the Visual Quantum Mechanics course for high school and introductory college classes. Materials inlcude tutorial activities in concepts of energy diagrams, probability, and wavefunctions, and some computer activities.

Axmann, Wally; Group, Kansas S.

2004-04-04

366

A mechanics baseline test  

NSDL National Science Digital Library

Reports the design of the "Mechanics Baseline Test," an instrument to assess students' understandings about concepts in mechanics. Discusses how comparisons of test results with extensive baseline data can be used to evaluate instruction at all levels. Includes a copy of the instrument.

Hestenes, David; Wells, Malcolm

2005-10-27

367

What is quantum mechanics?  

Microsoft Academic Search

We discuss the arguments for suspecting that there exists a classical, i.e. deterministic theory underlying quantum mechanics. A difficulty is that an explanation must be found of the fact that the Hamiltonian, which is defined to be the operator that generates evolution in time, is bounded from below. The mechanism that can produce exactly such a constraint is identified in

Gerard't Hooft

2007-01-01

368

Engineer Equipment Mechanic.  

ERIC Educational Resources Information Center

Developed as part of the Marine Corps Institute (MCI) correspondence training program, this course on engineer equipment mechanics is designed to advance the professional competence of privates through sergeants as equipment mechanics, Military Occupation Specialty 1341, and is adaptable for nonmilitary instruction. Introductory materials include…

Marine Corps Inst., Washington, DC.

369

Yale University Mechanical Engineering  

E-print Network

) ­ #92474A029 (4x) #12;OpenHand Yale University Mechanical Engineering 3D Printer Requirements · Current · Majority of parts are designed to not require support material · Authors do not know how well alternate 3D printers will produce adequate components #12;OpenHand Yale University Mechanical Engineering Finger

Dollar, Aaron M.

370

Discrete dark matter mechanism  

E-print Network

We present the Discrete Dark Matter mechanism (DDM) which consist on the stability of the dark matter from a spontaneous breaking of a flavor symmetry group into one of its subgroups. Here we present the inclusion of the quarks on this mechanism promoting the quarks to transform non-trivial under the flavor group.

E. Peinado

2012-10-25

371

Deformation Mechanisms and Microstructures  

NSDL National Science Digital Library

In this exercise, students match microstructures to the deformation mechanisms by which they form; compare pairs of photomicrographs chosen to highlight key differences between some common microstructures; and complete a self-quiz in which they identify microstructures and infer deformation mechanisms from photomicrographs.

Ormand, Carol

372

INJECTION WELL MECHANICAL INTEGRITY  

EPA Science Inventory

EPA's underground injection control regulations require that all injection wells demonstrate mechanical integrity. The regulations state that an injection well has mechanical integrity if (1) there is no significant leak in the casing, tubing or packer and (2) there is no signifi...

373

The Clementine mechanisms  

NASA Technical Reports Server (NTRS)

The Clementine spacecraft was developed under the 'faster, better, cheaper' theme. The constraints of a low budget coupled with an unusually tight schedule forced many departures from the normal spacecraft development methods. This paper discusses technical lessons learned about several of the mechanisms on the Clementine spacecraft as well as managerial lessons learned for the entire mechanisms subsystem. A quick overview of the Clementine mission is included; the mission schedule and environment during the mechanisms releases and deployment are highlighted. This paper then describes the entire mechanisms subsystem. The design and test approach and key philosophies for a fast-track program are discussed during the description of the mechanisms subsystem. The mechanism subsystem included a marman clamp separation system, a separation nut separation system, a solar panel deployment and pointing system, a high gain antenna feed deployment system, and two separate sensor cover systems. Each mechanism is briefly discussed. Additional technical discussion is given on the marman clamp design, the sensor cover designs, and the design and testing practices for systems driven by heated actuators (specifically paraffin actuators and frangibolts). All of the other mechanisms were of conventional designs and will receive less emphasis. Lessons learned are discussed throughout the paper as they applied to the systems being discussed. Since there is information on many different systems, this paper is organized so that information on a particular topic can be quickly referenced.

Purdy, William; Hurley, Michael

1995-01-01

374

Local mucociliary defence mechanisms  

Microsoft Academic Search

The lung is continually at risk of exposure to noxious environmental agents and respiratory pathogens. An elaborate series of defence mechanisms have been developed to protect the airways from these insults. The lower respiratory tract is protected by local mucociliary mechanisms that involve the integration of the ciliated epithelium, periciliary fluid and mucus. Mucus acts as a physical and chemical

M. A. Chilvers; C. O’Callaghan

2000-01-01

375

Biomimetic mechanically adaptive nanocomposites  

Microsoft Academic Search

The development of a new class of mechanically adaptive nanocomposites has been inspired by biological creatures such as sea cucumbers, which have the ability to reversibly change the stiffness of their dermis. Several recent studies have related this dynamic mechanical behaviour to the distinctive nanocomposite architecture of the collagenous tissue, in which interactions among rigid collagen fibrils, embedded in a

Kadhiravan Shanmuganathan; Jeffrey R. Capadona; Stuart J. Rowan; Christoph Weder

2010-01-01

376

Geometrization of Quantum Mechanics  

E-print Network

We show that it is possible to represent various descriptions of Quantum Mechanics in geometrical terms. In particular we start with the space of observables and use the momentum map associated with the unitary group to provide an unified geometrical description for the different pictures of Quantum Mechanics. This construction provides an alternative to the usual GNS construction for pure states.

J. F. Carinena; J. Clemente-Gallardo; G. Marmo

2007-01-19

377

Bacteriophage resistance mechanisms  

Microsoft Academic Search

Phages are now acknowledged as the most abundant microorganisms on the planet and are also possibly the most diversified. This diversity is mostly driven by their dynamic adaptation when facing selective pressure such as phage resistance mechanisms, which are widespread in bacterial hosts. When infecting bacterial cells, phages face a range of antiviral mechanisms, and they have evolved multiple tactics

Simon J. Labrie; Julie E. Samson; Sylvain Moineau

2010-01-01

378

Mechanisms (session summary)  

Microsoft Academic Search

Takuya Katayama lead the discussion on Mechanisms for Software Process Description. He classified software process views as reflected in the position papers into functional, behavioral and enactional, described some generally desirable properties of software process mechanisms, and listed those projects that seemed to have obtained non-toy experience.He made a plea for each of these projects to answer a questionnaire that,

Gail E. Kaiser

1990-01-01

379

Mechanisms and Psychological Explanation  

Microsoft Academic Search

As much as assumptions about mechanisms and mechanistic explanation have deeply affected psychology, they have received disproportionately little analysis in philosophy. After a historical survey of the influences of mechanistic approaches to explanation of psychological phenomena, we specify the nature of mechanisms and mechanistic explanation. Contrary to some treatments of mechanistic explanation, we maintain that explanation is an epistemic activity

Cory Wright; William Bechtel

380

Morphological Evolution: Epigenetic Mechanisms  

E-print Network

and mechanically excitable materials, and later other conditional form-generating processes, such as tissue been generated by the highly integrated developmental programmes charac- teristic of modern. These mechanisms are responsible for continued generation of morpho- logical novelty, and are ultimately involved

Müller, Gerd B.

381

Mechanics: Ideas, problems, applications  

Microsoft Academic Search

The book contains the published articles and reports by academician Ishlinskii which deal with the concepts and ideas of modern mechanics, its role in providing a general understanding of the natural phenomena, and its applications to various problems in science and engineering. Attention is given to the methodological aspects of mechanics, to the history of the theories of plasticity, friction,

A. Iu. Ishlinskii

1985-01-01

382

Mechanical engineering Department Seminar  

E-print Network

Mechanical engineering Department Seminar Alexis Sauer-Budge Senior Research Scientist, Fraunhofer of mechanical engineering and life sciences 11:00 AM Friday, April 27th, 2012 Room 245, 110 Cummington Street CMI Adjunct Research Assistant Professor, BME, Boston University Innovations at the intersection

383

Seesaw mechanism and leptogenesis  

E-print Network

A brief overview of the phenomenology related to the seesaw mechanism and the baryogenesis via leptogenesis is presented. In particular, it is explained how large but not maximal lepton mixing can be achieved within the type II seesaw mechanism. Moreover, the consequences for leptogenesis are explored, including flavor effects.

D. Falcone

2006-12-05

384

The Mechanization of Work.  

ERIC Educational Resources Information Center

Discusses how mechanization of work has been treated by economists, what its effect has been on the past U.S. economy, and what its future effect is likely to be. Emphasizes the impact of mechanization on the shifting structure and character of the labor force and evolution of the work environment. (Author/JN)

Ginzberg, Eli

1982-01-01

385

Mechanisms in biomedical ontology  

PubMed Central

The concept of a mechanism has become a standard proposal for explanations in biology. It has been claimed that mechanistic explanations are appropriate for systems biology, because they occupy a middle ground between strict reductionism and holism. Because of their importance in the field a formal ontological description of mechanisms is desirable. The standard philosophical accounts of mechanisms are often ambiguous and lack the clarity that can be provided by a formal-ontological framework. The goal of this paper is to clarify some of these ambiguities and suggest such a framework for mechanisms. Taking some hints from an "ontology of devices" I suggest as a general approach for this task the introduction of functional kinds and functional parts by which the particular relations between a mechanism and its components can be captured. PMID:23046727

2012-01-01

386

Membrane Quantum Mechanics  

E-print Network

We consider the multiple M2-branes wrapped on a compact Riemann surface and study the arising quantum mechanics by taking the limit where the size of the Riemann surface goes to zero. The IR quantum mechanical models resulting from the BLG-model and the ABJM-model compactified on a torus are N = 16 and N = 12 superconformal gauged quantum mechanics. After integrating out the auxiliary gauge fields we find OSp(16|2) and SU(1,1|6) quantum mechanics from the reduced systems. The curved Riemann surface is taken as a holomorphic curve in a Calabi-Yau space to preserve supersymmetry and we present a prescription of the topological twisting. We find the N = 8 superconformal gauged quantum mechanics that may describe the motion of two wrapped M2-branes in a K3 surface.

Okazaki, Tadashi

2014-01-01

387

Structural Mechanics & Solid Mechanics A finite element toolbox to MATLAB  

E-print Network

Structural Mechanics & Solid Mechanics Department of Mechanics and Materials CALFEM A finite 1999 c Copyright 1992­99 by the Division of Structural Mechanics and the Department of Solid Mechanics The Department of Solid Mechanics Lund University PO Box 118 S­221 00 Lund SWEDEN Phone: +46 46 222 0000 Fax: +46

Ehrhardt, Matthias

388

MECHANICAL ENGINEERING Program of Study  

E-print Network

MECHANICAL ENGINEERING Program of Study Correspondence The Department of Mechanical Engineering offers graduate programs in the fields of thermal science and engineering mechanics. Current areas of research activity include Biomedical Engineering, Biomimetics, Composite Materials, Computational Mechanics

Thomas, Andrew

389

Integrated Mechanical & Electrical Engineering (IMEE)  

E-print Network

Integrated Mechanical & Electrical Engineering (IMEE) Department of Electronic & Electrical Engineering and Department of Mechanical Engineering #12;Graduates able to work in both mechanical of Mechanical Engineers (IMechE) n Develop essential engineering skills through extensive project work n Enhance

Burton, Geoffrey R.

390

MECHANICAL ENGINEERING UNDERGRADUATE HONORS THESIS  

E-print Network

MECHANICAL ENGINEERING UNDERGRADUATE HONORS THESIS The Department of Mechanical Engineering offers a program leading to a Bachelor of Science in Mechanical Engineering with Honors. This program provides a unique opportunity for qualified mechanical engineering majors to conduct independent study

Prinz, Friedrich B.

391

Statistical Mechanics with focus on  

E-print Network

Statistical Mechanics with focus on Liquids, Solutions and Colloidal Systems Course contents A. Foundations of statistical mechanics Classical dynamics ­ Hamilton's and Liouville's equations The concept thermodynamics and statistical mechanics. B. Liquid state theory; Equilibrium statistical mechanics for liquids

Johannesson, Henrik

392

Quantum Mechanics From the Cradle?  

ERIC Educational Resources Information Center

States that the major problem in learning quantum mechanics is often the student's ignorance of classical mechanics and that one conceptual hurdle in quantum mechanics is its statistical nature, in contrast to the determinism of classical mechanics. (MLH)

Martin, John L.

1974-01-01

393

Overview of Bohmian Mechanics  

E-print Network

This chapter provides a comprehensive overview of the Bohmian formulation of quantum mechanics. It starts with a historical review of the difficulties found by Louis de Broglie, David Bohm, and John S. Bell to convince the scientific community about the validity and utility of Bohmian mechanics. Then, a formal explanation of Bohmian mechanics for nonrelativistic, single-particle quantum systems is presented. The generalization to many-particle systems, where the exchange interaction and the spin play an important role, is also presented. After that, the measurement process in Bohmian mechanics is discussed. It is emphasized that Bohmian mechanics exactly reproduces the mean value and temporal and spatial correlations obtained from the standard, that is the Copenhagen or orthodox, formulation. The ontological characteristics of Bohmian mechanics provide a description of measurements as another type of interaction without the need for introducing the wave function collapse. Several solved problems are presented at the end of the chapter, giving additional mathematical support to some particular issues. A detailed description of computational algorithms to obtain Bohmian trajectories from the numerical solution of the Schrodinger or the Hamilton-Jacobi equations are presented in an appendix. The motivation of this chapter is twofold: first, as a didactic introduction to Bohmian formalism, which is used in the subsequent chapters, and second, as a self-contained summary for any newcomer interested in using Bohmian mechanics in his or her daily research activity.

Xavier Oriols; Jordi Mompart

2012-06-05

394

Tethered satellite control mechanism  

NASA Technical Reports Server (NTRS)

The tethered satellite control mechanisms consist of four major subsystems. The reel drive mechanism stores the tether. It is motor driven and includes a level wind to uniformly feed the tether to the reel. The lower boom mechanism serves two primary functions: (1) it measures tether length and velocity as the tether runs through the mechanism, and (2) it reads the tether tension at the reel. It also provides change the direction for the tether from the reel to the upper boom mechanism. The deployment boom positions the upper boom mechanism with satellite out of the cargo bay. The deployment function places the 500-kg satellite 20 m away from the Space Shuttle (producing a small natural gravity gradient force), impacts an initial velocity to the satellite for deployment, and allows for satellite docking at a safe distance from the body of the Space Shuttle. The upper boom mechanism (UBM) services three functions: (1) it provides tether control to the satellite as the satellite swings in and out of plane; (2) it reads tether tension in the low range during the early deployment and final retrieval parts of the mission; and (3) it produces additional tether tension at the reel when tether tension to the satellite is in the low range.

Kyrias, G. M.

1983-01-01

395

The Mechanisms of Involuntary Attention  

ERIC Educational Resources Information Center

We tested 3 mechanisms of involuntary attention: (1) a perceptual enhancement mechanism, (2) a response-decision mechanism, and (3) a serial-search mechanism. Experiment 1 used a response deadline technique to compare the perceptual enhancement and the decision mechanisms and found evidence consistent with the decision mechanism. Experiment 2 used…

Prinzmetal, William; Ha, Ruby; Khani, Aniss

2010-01-01

396

07SCHOOL OF MECHANICAL ENGINEERING  

E-print Network

07SCHOOL OF MECHANICAL ENGINEERING UNDERGRADUATE DEGREES School of Mechanical Engineering FACULTY OF ENGINEERING Undergraduate Degrees 2015 #12;www.engineering.leeds.ac.uk/mechanical UNDERGRADUATE DEGREES SCHOOL OF MECHANICAL ENGINEERING The School of Mechanical Engineering offers both a broad mechanical engineering degree

Dimitrova, Vania

397

Is quantum mechanics exact?  

SciTech Connect

We formulate physically motivated axioms for a physical theory which for systems with a finite number of degrees of freedom uniquely lead to quantum mechanics as the only nontrivial consistent theory. Complex numbers and the existence of the Planck constant common to all systems arise naturally in this approach. The axioms are divided into two groups covering kinematics and basic measurement theory, respectively. We show that even if the second group of axioms is dropped, there are no deformations of quantum mechanics which preserve the kinematic axioms. Thus, any theory going beyond quantum mechanics must represent a radical departure from the usual a priori assumptions about the laws of nature.

Kapustin, Anton [California Institute of Technology, Pasadena, California 91125 (United States)] [California Institute of Technology, Pasadena, California 91125 (United States)

2013-06-15

398

Rotary mechanical latch  

DOEpatents

A rotary mechanical latch for positive latching and unlatching of a rotary device with a latchable rotating assembly having a latching gear that can be driven to latched and unlatched states by a drive mechanism such as an electric motor. A cam arm affixed to the latching gear interfaces with leading and trailing latch cams affixed to a flange within the drive mechanism. The interaction of the cam arm with leading and trailing latch cams prevents rotation of the rotating assembly by external forces such as those due to vibration or tampering.

Spletzer, Barry L.; Martinez, Michael A.; Marron, Lisa C.

2012-11-13

399

Advanced Mechanics. Mathematical Introduction  

E-print Network

Classical non-relativistic mechanics in a general setting of time-dependent transformations and reference frame changes is formulated in the terms of fibre bundles over the time-axis R. Connections on fibre bundles are the main ingredient in this formulation of mechanics which thus is covariant under reference frame transformations. The basic notions of a non-relativistic reference frame, a relative velocity, a free motion equation, a relative acceleration, an external force are formulated. Newtonian, Lagrangian, Hamiltonian mechanical systems and the relations between them are defined. Lagrangian and Hamiltonian conservation laws are considered.

G. Giachetta; L. Mangiarotti; G. Sardanashvily

2009-11-03

400

Mechanics: Ideas, problems, applications  

NASA Astrophysics Data System (ADS)

The book contains the published articles and reports by academician Ishlinskii which deal with the concepts and ideas of modern mechanics, its role in providing a general understanding of the natural phenomena, and its applications to various problems in science and engineering. Attention is given to the methodological aspects of mechanics, to the history of the theories of plasticity, friction, gyroscopic and inertial systems, and inertial navigation, and to mathematical methods in mechanics. The book also contains essays on some famous scientists and engineers.

Ishlinskii, A. Iu.

401

Mechanism of Gravity Impulse  

E-print Network

It is well-known that energy-momentum is the source of gravitational field. For a long time, it is generally believed that only stars with huge masses can generate strong gravitational field. Based on the unified theory of gravitational interactions and electromagnetic interactions, a new mechanism of the generation of gravitational field is studied. According to this mechanism, in some special conditions, electromagnetic energy can be directly converted into gravitational energy, and strong gravitational field can be generated without massive stars. Gravity impulse found in experiments is generated by this mechanism.

Ning Wu

2005-10-01

402

In Vitro Antitumor Mechanism of (E)-N-(2-methoxy-5-(((2,4,6-trimethoxystyryl)sulfonyl)methyl)pyridin-3-yl)methanesulfonamide.  

PubMed

ON01910.Na [sodium (E)-2-(2-methoxy-5-((2,4,6-trimethoxystyrylsulfonyl)methyl)phenylamino)acetate; Rigosertib, Estybon], a styryl benzylsulfone, is a phase III stage anticancer agent. This non-ATP competitive kinase inhibitor has multitargeted activity, promoting mitotic arrest and apoptosis. Extensive phase I/II studies with ON01910.Na, conducted in patients with solid tumors and hematologic cancers, demonstrate excellent efficacy. However, issues remain affecting its development. These include incomplete understanding of antitumor mechanisms, low oral bioavailability, and unpredictable pharmacokinetics. We have identified a novel (E)-styrylsulfonyl methylpyridine [(E)-N-(2-methoxy-5-((2,4,6-trimethoxystyrylsulfonyl)methyl)pyridin-3-yl)methanesulfonamide (TL-77)] which has shown improved oral bioavailability compared with ON01910.Na. Here, we present detailed cellular mechanisms of TL-77 in comparison with ON01910.Na. TL-77 displays potent growth inhibitory activity in vitro (GI50 < 1?M against HCT-116 cells), demonstrating 3- to 10-fold greater potency against tumor cell lines when compared with normal cells. Cell-cycle analyses reveal that TL-77 causes significant G2/M arrest in cancer cells, followed by the onset of apoptosis. In cell-free conditions, TL-77 potently inhibits tubulin polymerization. Mitotically arrested cells display multipolar spindles and misalignment of chromosomes, indicating that TL-77 interferes with mitotic spindle assembly in cancer cells. These effects are accompanied by induction of DNA damage, inhibition of Cdc25C phosphorylation [indicative of Plk1 inhibition], and downstream inhibition of cyclin B1. However, kinase assays failed to confirm inhibition of Plk1. Nonsignificant effects on phosphoinositide 3-kinase/Akt signal transduction were observed after TL-77 treatment. Analysis of apoptotic signaling pathways reveals that TL-77 downregulates expression of B-cell lymphoma 2 family proteins (Bid, Bcl-xl, and Mcl-1) and stimulates caspase activation. Taken together, TL-77 represents a promising anticancer agent worthy of further evaluation. PMID:25316768

Lu, Tiangong; Laughton, Charles A; Wang, Shudong; Bradshaw, Tracey D

2015-01-01

403

The kinase DYRK1A phosphorylates the transcription factor FKHR at Ser329 in vitro, a novel in vivo phosphorylation site.  

PubMed Central

Forkhead in rhabdomyosarcoma (FKHR) is a transcription factor that has been implicated in the control of gene expression by insulin, as well as the regulation of apoptosis by survival factors. These signals trigger the protein kinase B (PKB)-catalysed phosphorylation of FKHR at three residues (Thr(24), Ser(256) and Ser(319)) by a phosphoinositide 3-kinase-dependent pathway that results in the nuclear exit and inactivation of this transcription factor. Here, we have identified a conserved residue (Ser(329)) as a novel in vivo phosphorylation site on FKHR. Ser(329) phosphorylation also decreases the ability of FKHR to stimulate gene transactivation and reduces the proportion of FKHR present in the nucleus. However, unlike the residues targetted by PKB, Ser(329) is phosphorylated in unstimulated HEK-293 cells, and phosphorylation is not increased by stimulation with insulin-like growth factor-1 or by transfection with 3-phosphoinositide-dependent protein kinase-1. We have also purified a protein kinase to near homogeneity from rabbit skeletal muscle that phosphorylates FKHR at Ser(329) specifically and identified it as DYRK1A (dual-specificity tyrosine-phosphorylated and regulated kinase 1A). We find that FKHR and DYRK1A co-localize in discrete regions of the nucleus and can be co-immunoprecipitated from cell extracts. These experiments suggest that DYRK1A may phosphorylate FKHR at Ser(329) in vivo. PMID:11311120

Woods, Y L; Rena, G; Morrice, N; Barthel, A; Becker, W; Guo, S; Unterman, T G; Cohen, P

2001-01-01

404

Regulation of osteoclasts by membrane-derived lipid mediators.  

PubMed

Osteoclasts are bone-resorbing cells of monocytic origin. An imbalance between bone formation and resorption can lead to osteoporosis or osteopetrosis. Osteoclastogenesis is triggered by RANKL- and IP3-induced Ca(2+) influx followed by activation of NFATc1, a master transcription factor for osteoclastogenic gene regulation. During differentiation, osteoclasts undergo cytoskeletal remodeling to migrate and attach to the bone surface. Simultaneously, they fuse with each other to form multinucleated cells. These processes require PI3-kinase-dependent cytoskeletal protein activation to initiate cytoskeletal remodeling, resulting in the formation of circumferential podosomes and fusion-competent protrusions. In multinucleated osteoclasts, circumferential podosomes mature into stabilized actin rings, which enables the formation of a ruffled border where intensive membrane trafficking is executed. Membrane lipids, especially phosphoinositides, are key signaling molecules that regulate osteoclast morphology and act as second messengers and docking sites for multiple important effectors. We examine the critical roles of phosphoinositides in the signaling cascades that regulate osteoclast functions. PMID:23296124

Oikawa, Tsukasa; Kuroda, Yukiko; Matsuo, Koichi

2013-09-01

405

Mechanism of mechanical fatigue of silica glass  

SciTech Connect

Research continued on the mechanism of mechanical fatigue of silica glass. Significant accomplishments this quarter include: strength increase of abraded glasses by thermal and chemical treatments was found best explained by crack tip blunting; fatigue was found to involve crack initiation in addition to crack propagation, and only water and ammonia were found capable of causing crack initiation from blunt crack tips at sub-critical stresses; water was found to enter into silica glass during microhardness indentation as well as during slow crack growth in water-containing atmospheres at room temperature; water diffusion at low temperature showed anomalous phenomena including initial time dependences of solubility and apparent diffusion coefficient; glass containing water showed greater fatigue susceptibility and lower crack initiation stress than dry glass; fatigue of abraded silica glass showed effects of cyclic stress when tested by the rotation bending method while no cyclic stress effect was observed in reciprocal bending; and a new coating which can reduce fatigue of silica glass was developed.

Tomozawa, Minoru.

1990-01-01

406

Quantum Mechanics II (Undergraduate)  

E-print Network

, and applications of quantum mechanics to materials science/solid-state physics. Grades: Homework: 15%, Midertm: 40 other selected topics from quantum information (see the QUNET reference) and solid-state physics. All

Nickrent, Daniel L.

407

Brain Mechanisms of Movement.  

ERIC Educational Resources Information Center

This article focuses on the mechanisms of the human brain which bring about body movements. Included is a discussion of the way in which the brain and spinal cord issue commands and receive signals. (Author/SA)

Evarts, Edward V.

1979-01-01

408

MECHANISMS OF PESTICIDE DEGRADATION  

EPA Science Inventory

This research project was initiated with the overall objective of determining (1) the chemical structures of toxic components of toxaphene, (2) to study anaerobic metabolism to degrade toxaphene and other pesticides, and (3) to understand toxic action mechanism of chlordimeform. ...

409

Applied Mechanics Materials  

NSDL National Science Digital Library

This site from SpaceTEC National Aerospace Technical Education Center presents materials on applied mechanics in the aerospace workplace. Topics include machine shop safety, non-cutting hand tools, cutting hand tools, drilling, measurement, basic calculations, hardware and blueprints.

2010-10-18

410

Playing at Statistical Mechanics  

ERIC Educational Resources Information Center

Discussed are the applications of counting techniques of a sorting game to distributions and concepts in statistical mechanics. Included are the following distributions: Fermi-Dirac, Bose-Einstein, and most probable. (RH)

Clark, Paul M.; And Others

1974-01-01

411

Ultralight, ultrastiff mechanical metamaterials  

E-print Network

The mechanical properties of ordinary materials degrade substantially with reduced density because their structural elements bend under applied load. We report a class of microarchitected materials that maintain a nearly ...

Zheng, Xiaoyu

412

Mechanical sensors and actuators  

Microsoft Academic Search

A review on mechanical sensing techniques based on magnetic methods is presented, with special focus on strain sensing in civil engineering. Some examples and features of magnetic actuation with giant magnetostrictive and magnetic shape memory alloys will be shown.

M. Pasquale

2003-01-01

413

Introduction to Quantum Mechanics  

NSDL National Science Digital Library

This text is intended for junior/senior Quantum Mechanics courses. It covers the fundamentals of quantum theory in a concise manner, covering topics from the basic formalism through perturbation theory, the adiabatic approximation, and scattering.

Griffiths, David J.

2005-04-16

414

Essays on mechanism design  

E-print Network

This thesis is a collection of three essays on mechanism design. In Chapter 1, we consider a general Informed Principal Problem in the context of procurement. Both the potential suppliers and the buyer hold some private ...

Balestrieri, Filippo

2008-01-01

415

Mechanisms of catalyst deactivation  

Microsoft Academic Search

The literature treating mechanisms of catalyst deactivation is reviewed. Intrinsic mechanisms of catalyst deactivation are many; nevertheless, they can be classified into six distinct types: (i) poisoning, (ii) fouling, (iii) thermal degradation, (iv) vapor compound formation accompanied by transport, (v) vapor-solid and\\/or solid-solid reactions, and (vi) attrition\\/crushing. As (i), (iv), and (v) are chemical in nature and (ii) and (v)

Calvin H Bartholomew

2001-01-01

416

Evaluation of Mechanical Properties  

Microsoft Academic Search

\\u000a Mechanical properties such as elastic modulus, fracture stress, and yield stress of nano\\/micromaterials are fundamental data\\u000a for practical design of nano\\/micromaterial-based devices. These properties generally differ from those of bulk material because\\u000a of size effects. This chapter is devoted to an introduction of some techniques for evaluating the mechanical properties of\\u000a nanowires and thin wires. In order to clarify the

Mikio Muraoka; Hironori Tohmyoh

417

Space Mechanisms Technology Workshop  

NASA Technical Reports Server (NTRS)

The Mechanical Components Branch at NASA Glenn Research Center hosted a workshop to discuss the state of drive systems technology needed for space exploration. The Workshop was held Thursday, November 2, 2000. About 70 space mechanisms experts shared their experiences from working in this field and considered technology development that will be needed to support future space exploration in the next 10 to 30 years.

Oswald, Fred B. (Editor)

2001-01-01

418

Mechanics in a wheelchair  

NASA Astrophysics Data System (ADS)

It is not only possible for a student sitting in a wheelchair to participate in many active engagement activities. Using a wheelchair is also of extra value for non-physically disabled students in many experiential mechanics activities since the friction is low and kinesthetic experience involved. A wheelchair can also be used as an engaging "tool" for connecting mechanical concepts and real-world phenomena.

Bernhard, Karin; Bernhard, Jonte

1999-12-01

419

Constructibility in Quantum Mechanics  

E-print Network

We propose a set theoretical foundation including an axiom of constructibility and derive a generalized quantum mechanics by postulating symmetry of action. The Schroedinger equation proves to be a special case of a nonlinear sigma model. Quantum mechanics is obtained here without the requirement for presupposing the statistical interpretation of the wave function; thus this derivation becomes continuous with prior physics. In this theory we also see space-time as relational and the fields as free of singularities.

D. J. Bendaniel

2008-06-06

420

Supersymmetry in quantum mechanics  

Microsoft Academic Search

In the past ten years, the ideas of supersymmetry have been profitably applied to many nonrelativistic quantum mechanical\\u000a problems. In particular, there is now a much deeper understanding of why certain potentials are analytically solvable. In\\u000a this lecture I review the theoretical formulation of supersymmetric quantum mechanics and discuss many of its applications.\\u000a I show that the well-known exactly solvable

Avinash Khare

1997-01-01

421

Graduate Quantum Mechanics Reform  

NSDL National Science Digital Library

We address four main areas in which graduate quantum mechanics education can be improved: course content, textbook, teaching methods, and assessment tools. We report on a three year longitudinal study at the Colorado School of Mines using innovations in all these areas. In particular, we have modified the content of the course to reflect progress in the field of quantum mechanics over the last 50 years, used textbooks that include such content, incorporated a variety of teaching techniques based on physics education research, and used a variety of assessment tools to study the effectiveness of these reforms. We present a new assessment tool, the Graduate Quantum Mechanics Conceptual Survey, and further testing of a previously developed assessment tool, the Quantum Mechanics Conceptual Survey. We find that graduate students respond well to research-based techniques that have been tested mainly in introductory courses, and that they learn much of the new content introduced in each version of the course. We also find that students' ability to answer conceptual questions about graduate quantum mechanics is highly correlated with their ability to solve calculational problems on the same topics. In contrast, we find that students' understanding of basic undergraduate quantum mechanics concepts at the modern physics level is not improved by instruction at the graduate level.

Carr, Lincoln D.; Mckagan, Sam B.

2009-05-06

422

Lung Parenchymal Mechanics  

PubMed Central

The lung parenchyma comprises a large number of thin-walled alveoli, forming an enormous surface area, which serves to maintain proper gas exchange. The alveoli are held open by the transpulmonary pressure, or prestress, which is balanced by tissues forces and alveolar surface film forces. Gas exchange efficiency is thus inextricably linked to three fundamental features of the lung: parenchymal architecture, prestress, and the mechanical properties of the parenchyma. The prestress is a key determinant of lung deformability that influences many phenomena including local ventilation, regional blood flow, tissue stiffness, smooth muscle contractility, and alveolar stability. The main pathway for stress transmission is through the extracellular matrix. Thus, the mechanical properties of the matrix play a key role both in lung function and biology. These mechanical properties in turn are determined by the constituents of the tissue, including elastin, collagen, and proteoglycans. In addition, the macroscopic mechanical properties are also influenced by the surface tension and, to some extent, the contractile state of the adherent cells. This article focuses on the biomechanical properties of the main constituents of the parenchyma in the presence of prestress and how these properties define normal function or change in disease. An integrated view of lung mechanics is presented and the utility of parenchymal mechanics at the bedside as well as its possible future role in lung physiology and medicine are discussed. PMID:23733644

Suki, Bela; Stamenovic, Dimitrije; Hubmayr, Rolf

2014-01-01

423

Mechanics of the Orbita  

PubMed Central

The oculomotor periphery was formerly regarded as a simple mechanism executing complex behaviors explicitly specified by innervation. It is now recognized that several fundamental aspects of ocular motility are properties of the extraocular muscles (EOMs) and their associated connective tissue pulleys. The Active Pulley Hypothesis proposes that rectus and inferior oblique EOMs have connective tissue soft pulleys that are actively controlled by the direction action of the EOMs’ orbital layers. Functional imaging and histology have suggested that the rectus pulley array constitutes an inner mechanism, similar to a gimbal, that is rotated torsionally around the orbital axis by an outer mechanism driven by the oblique EOMs. This arrangement may mechanically account for several commutative aspects of ocular motor control, including Listing’s law, yet permits implementation of noncommutative motility as during the vestibulo-ocular reflex. Recent human behavioral studies, as well neurophysiology in monkeys, are consistent with mechanical rather than central neural implementation of Listing’s law. Pathology of the pulley system is associated with predictable patterns of strabismus that are surgically treatable when the pathologic anatomy is characterized by imaging. This mechanical determination may imply limited possibilities for neural adaptation to some ocular motor pathologies, but indicates greater potential for surgical treatments. PMID:17314483

Demer, Joseph L.

2008-01-01

424

Wear and Tear - Mechanical  

NASA Technical Reports Server (NTRS)

The focus of this chapter is on the long term wear and tear, or aging, of the mechanical subsystem of a spacecraft. The mechanical subsystem is herein considered to be the primary support structure (as in a skeleton or exoskeleton) upon which all other spacecraft systems rest, and the associated mechanisms. Mechanisms are devices which have some component that moves at least once, in response to some type of passive or active control system. For the structure, aging may proceed as a gradual degradation of mechanical properties and/or function, possibly leading to complete structural failure over an extended period of time. However, over the 50 years of the Space Age such failures appear to be unusual. In contrast, failures for mechanisms are much more frequent and may have a very serious effect on mission performance. Just as on Earth, all moving devices are subject to normal (and possibly accelerated) degradation from mechanical wear due to loss or breakdown of lubricant, misalignment, temperature cycling effects, improper design/selection of materials, fatigue, and a variety of other effects. In space, such environmental factors as severe temperature swings (possibly 100's of degrees C while going in and out of direct solar exposure), hard vacuum, micrometeoroids, wear from operation in a dusty or contaminated environment, and materials degradation from radiation can be much worse. In addition, there are some ground handling issues such as humidity, long term storage, and ground transport which may be of concern. This chapter addresses the elements of the mechanical subsystem subject to wear, and identifies possible causes. The potential impact of such degradation is addressed, albeit with the recognition that the impact of such wear often depends on when it occurs and on what specific components. Most structural elements of the mechanical system typically are conservatively designed (often to a safety factor of greater than approximately 1.25 on yield for unmanned spacecraft) but do not have backup structure due to the added mass this would impose, and also due to the fact that structural elements can be accurately modeled mathematically and in test. Critical mechanisms or devices may have backups, or alternate work-arounds, since characterization of these systems in a 1g environment is less accurate than structure, and repair in-space is often impossible.

Swanson, Theodore

2008-01-01

425

Mechanical Engineering Is Mechanical Engineering right for me?  

E-print Network

Mechanical Engineering Is Mechanical Engineering right for me? If you are interested in the wide range of principles related to mechanical systems then Mechanical Engineering is well suited to you. A Mechanical Engineering degree programme will focus on aspects such as analysis, design, manufacture

Harman, Neal.A.

426

Consider mechanical recompression evaporation  

SciTech Connect

Mechanical recompression is a technique for improving latent heat management by recycling the latent heat of evaporation. The product vapor is isotropically compressed to a higher temperature and pressure so that it can be used as the heating medium that drives the evaporation. The mechanical recompression evaporator typically requires no main process steam, cooling water, or process steam condenser. The economy is in the range of what could be achieved by a 10- to 20-effect evaporator. The technique of mechanical recompression evaporation is not wet. The chemical process industries (CPI) in North American and Europe began using it extensively in the 1970s, probably as a results of the oil crisis. And, it will continue to play a major role in the removal of water for process applications. This article explains what mechanical recompression evaporation is, its advantages and disadvantages, and where it can be used. It also provides guidance on selecting the evaporator and compressor--the most important components of a mechanical recompression evaporation system.

Ward, A.

1994-04-01

427

Conventional mechanical ventilation  

PubMed Central

The provision of mechanical ventilation for the support of infants and children with respiratory failure or insufficiency is one of the most common techniques that are performed in the Pediatric Intensive Care Unit (PICU). Despite its widespread application in the PICUs of the 21st century, before the 1930s, respiratory failure was uniformly fatal due to the lack of equipment and techniques for airway management and ventilatory support. The operating rooms of the 1950s and 1960s provided the arena for the development of the manual skills and the refinement of the equipment needed for airway management, which subsequently led to the more widespread use of endotracheal intubation thereby ushering in the era of positive pressure ventilation. Although there seems to be an ever increasing complexity in the techniques of mechanical ventilation, its successful use in the PICU should be guided by the basic principles of gas exchange and the physiology of respiratory function. With an understanding of these key concepts and the use of basic concepts of mechanical ventilation, this technique can be successfully applied in both the PICU and the operating room. This article reviews the basic physiology of gas exchange, principles of pulmonary physiology, and the concepts of mechanical ventilation to provide an overview of the knowledge required for the provision of conventional mechanical ventilation in various clinical arenas. PMID:20927268

Tobias, Joseph D.

2010-01-01

428

Mechanism of PDK1-catalyzed Thr-229 Phosphorylation of the S6K1 Protein Kinase*  

PubMed Central

PDK1 (phosphoinositide-dependent protein kinase-1) catalyzes phosphorylation of Thr-229 in the T-loop of S6K1?II (the 70-kDa 40 S ribosomal protein S6 kinase-1 ?II isoform), and Thr-229 phosphorylation is synergistic with C-terminal Thr-389 phosphorylation to activate S6K1?II regulatory functions in protein translation preinitiation complexes. Unlike its common AGC kinase subfamily member S6K1?II, PDK1 does not contain the synergistic C-terminal phosphorylation site, and it has been proposed that phosphorylated Thr-389 in S6K1?II may initially serve to trans-activate PDK1-catalyzed Thr-229 phosphorylation. Herein, we report direct binding and kinetic studies that showed PDK1 to exhibit nearly equal binding affinities and steady-state kinetic turnover numbers toward native (KdS6K1 = 1.2 ?m and kcat = 1.1 s?1) and the phosphomimicking T389E mutant S6K1?II (KdS6K1 = 1.5 ?m and kcat = 1.2 s?1), although ?2-fold enhanced specificity was displayed for the T389E mutant (kcat/KmS6K1 = 0.08 ?m?1 s?1 compared with 0.04 ?m?1 s?1). Considering that transient kinetic binding studies showed all nucleotide and S6K1?II substrates and products to rapidly associate with PDK1 (kon = 1–6 ?m?1 s?1), it was concluded that positioning a negative charge at residue Thr-389 reduced ?2-fold the occurrence of nonproductive binding events that precede formation of a reactive ternary complex for Thr-229 phosphorylation. In addition, steady-state kinetic data were most simply accommodated by an Ordered Bi Bi mechanism with competitive substrate inhibition, where (i) the initially formed PDK1-ATP complex phosphorylates the nucleotide-free form of the S6K1?II kinase and (ii) initial binding of S6K1?II precludes ATP binding to PDK1. PMID:19570988

Keshwani, Malik M.; Gao, Xinxin; Harris, Thomas K.

2009-01-01

429

Engineering Mechanics Annual Report 2001  

E-print Network

Engineering Mechanics Annual Report 2001 Graduate School Engineering Mechanics c/o Eindhoven.3 Outline of the field of Engineering Mechanics 1.2 1.4 Organization 1.3 1.5 Participants 1.4 1.6 Research Analysis 6.1 7. Engineering Mechanics (1) 7.1 8. Aerospace Structures and Computational Mechanics 8.1 9

Franssen, Michael

430

Engineering Mechanics Annual Report 2002  

E-print Network

Engineering Mechanics Annual Report 2002 Graduate School Engineering Mechanics c/o Eindhoven.1 1.2 Mission statement 1.2 1.3 Outline of the field of Engineering Mechanics 1.2 1.4 Organization 1 and Engineering 4.1 5. Engineering Mechanics (1) 5.1 6. Aerospace Structures and Computational Mechanics 6.1 7

Franssen, Michael

431

Particulate erosion mechanisms  

NASA Technical Reports Server (NTRS)

Particulate damage and erosion of ductile metals are today plaguing design and field engineers in diverse fields of engineering and technology. It was found that too many models and theories were proposed leading to much speculation from debris analysis and failure mechanism postulations. Most theories of solid particle erosion are based on material removal models which do not fully represent the actual physical processes of material removal. The various mechanisms proposed thus far are: melting, low-cycle fatigue, extrusion, delamination, shear localization, adhesive material transfer, etc. The experimental data on different materials highlighting the observed failure modes of the deformation and cutting wear processes using optical and scanning electron microscopy are presented. The most important mechanisms proved from the experimental observations of the specimens exposed to both spherical and angular particles are addressed, and the validity of the earlier theories discussed. Both the initial stages of damage and advanced stages of erosion were studied to gain a fundamental understanding of the process.

Veerabhadrarao, P.; Buckley, D. H.

1983-01-01

432

Epigenetics: Biology's Quantum Mechanics  

PubMed Central

The perspective presented here is that modern genetics is at a similar stage of development as were early formulations of quantum mechanics theory in the 1920s and that in 2010 we are at the dawn of a new revolution in genetics that promises to enrich and deepen our understanding of the gene and the genome. The interrelationships and interdependence of two views of the gene – the molecular biological view and the epigenetic view – are explored, and it is argued that the classical molecular biological view is incomplete without incorporation of the epigenetic perspective and that in a sense the molecular biological view has been evolving to include the epigenetic view. Intriguingly, this evolution of the molecular view toward the broader and more inclusive epigenetic view of the gene has an intriguing, if not precise, parallel in the evolution of concepts of atomic physics from Newtonian mechanics to quantum mechanics that are interesting to consider. PMID:22639577

Jorgensen, Richard A.

2011-01-01

433

Mechanisms of epigenetic memory.  

PubMed

Although genetics has an essential role in defining the development, morphology, and physiology of an organism, epigenetic mechanisms have an essential role in modulating these properties by regulating gene expression. During development, epigenetic mechanisms establish stable gene expression patterns to ensure proper differentiation. Such mechanisms also allow organisms to adapt to environmental changes and previous experiences can impact the future responsiveness of an organism to a stimulus over long timescales and even over generations. Here, we discuss the concept of epigenetic memory, defined as the stable propagation of a change in gene expression or potential induced by developmental or environmental stimuli. We highlight three distinct paradigms of epigenetic memory that operate on different timescales. PMID:24780085

D'Urso, Agustina; Brickner, Jason H

2014-06-01

434

Cnidarian internal stinging mechanism  

PubMed Central

Stinging mechanisms generally deliver venomous compounds to external targets. However, nematocysts, the microscopic stinging organelles that are common to all members of the phylum Cnidaria, occur and act in both external and internal tissue structures. This is the first report of such an internal piercing mechanism. This mechanism identifies prey items within the body cavity of the sea anemone and actively injects them with cytolytic venom compounds. Internal tissues isolated from sea anemones caused the degradation of live Artemia salina nauplii in vitro. When examined, the nauplii were found to be pierced by discharged nematocysts. This phenomenon is suggested to aid digestive phagocytic processes in a predator otherwise lacking the means to masticate its prey. PMID:19129118

Schlesinger, Ami; Zlotkin, Eliahu; Kramarsky-Winter, Esti; Loya, Y.

2008-01-01

435

MIRO Calibration Switch Mechanism  

NASA Technical Reports Server (NTRS)

The Jet Propulsion Laboratory has designed, analyzed, built, and tested a calibration switch mechanism for the MIRO instrument on the ROSETTA spacecraft. MIRO is the Microwave Instrument for the Rosetta Orbiter; this instrument hopes to investigate the origin of the solar system by studying the origin of comets. Specifically, the instrument will be the first to use submillimeter and millimeter wave heterodyne receivers to remotely examine the P-54 Wirtanen comet. In order to calibrate the instrument, it needs to view a hot and cold target. The purpose of the mechanism is to divert the instrument's field of view from the hot target, to the cold target, and then back into space. This cycle is to be repeated every 30 minutes for the duration of the 1.5 year mission. The paper describes the development of the mechanism, as well as analysis and testing techniques.

Suchman, Jason; Salinas, Yuki; Kubo, Holly

2001-01-01

436

Mechanisms in knockout reactions  

E-print Network

We report on the first detailed study of the mechanisms involved in knockout reactions, via a coincidence measurement of the residue and fast proton in one-proton knockout reactions, using the S800 spectrograph in combination with the HiRA detector array at the NSCL. Results on the reactions $^9$Be($^9$C,$^8$B+X)Y and $^9$Be($^8$B,$^7$Be+X)Y are presented. They are compared with theoretical predictions for both the diffraction and stripping reaction mechanisms, as calculated in the eikonal model. The data shows a clear distinction between the two reaction mechanisms, and the observed respective proportions are very well reproduced by the reaction theory. This agreement supports the results of knockout reaction analyses and their applications to the spectroscopy of rare isotopes.

D. Bazin; R. J. Charity; R. T. de Souza; M. A. Famiano; A. Gade; V. Henzl; D. Henzlova; S. Hudan; J. Lee; S. Lukyanov; W. G. Lynch; S. McDaniel; M. Mocko; A. Obertelli; A. M. Rogers; L. G. Sobotka; J. R. Terry; J. A. Tostevin; M. B. Tsang; M. S. Wallace

2009-02-16

437

Mechanisms of Neuropathic Pain  

PubMed Central

Neuropathic pain refers to pain that originates from pathology of the nervous system. Diabetes, infection (herpes zoster),nerve compression, nerve trauma, “channelopathies,” and autoimmune disease are examples of diseases that maycause neuropathic pain. The development ofbothanimal models and newer pharmacological strategies has led to an explosion of interest in the underlying mechanisms. Neuropathic pain reflects both peripheral and central sensitization mechanisms. Abnormal signals arise not only from injured axons but also from the intact nociceptors that share the innervation territory of the injured nerve. This review focuses on how both human studies and animal models are helping to elucidate the mechanisms underlying these surprisingly common disorders. The rapid gain in knowledge about abnormal signaling promises breakthroughs in the treatment of these often debilitating disorders. PMID:17015228

Campbell, James N.; Meyer, Richard A.

2007-01-01

438

Upper Airway Mechanics  

PubMed Central

This review discusses the pathophysiological aspects of sleep-disordered breathing, with focus on upper airway mechanics in obstructive and central sleep apnoea, Cheyne-Stokes respiration and obesity hypoventilation syndrome. These disorders constitute the end points of a spectrum with distinct yet interrelated mechanisms that lead to substantial pathology, i.e. increased upper airway collapsibility, control of breathing instability, increased work of breathing, disturbed ventilatory system mechanics and neurohormonal changes. Concepts are changing. Although sleep apnoea is considered more and more to be an increased loop gain disorder, the central type of apnoea is now considered as an obstructive event, because it causes pharyngeal narrowing, associated with prolonged expiration. Although a unifying concept for the pathogenesis is lacking, it seems that these patients are in a vicious circle. Knowledge of common patterns of sleep-disordered breathing may help to identify these patients and guide therapy. PMID:19478479

Verbraecken, Johan A.; De Backer, Wilfried A.

2009-01-01

439

The scar mechanism revisited  

E-print Network

Unstable periodic orbits are known to originate scars on some eigenfunctions of classically chaotic systems through recurrences causing that some part of an initial distribution of quantum probability in its vicinity returns periodically close to the initial point. In the energy domain, these recurrences are seen to accumulate quantum density along the