Sample records for factor usf2 binding
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Castro, Jazmin; Araya, Germán; Inostroza, Pamela; Hidalgo, Paulina; González-Ramos, Reinaldo; Sovino, Hugo; Boric, M Angélica; Fuentes, Ariel; Johnson, M Cecilia
2015-10-09
Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1. Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERα) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified. These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERα and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavity.
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Phosphorylation-dependent down-regulation of apolipoprotein A5 by insulin
DOE Office of Scientific and Technical Information (OSTI.GOV)
Nowak, Maxine; Helleboid-Chapman, Audrey; Jakel, Heidelinde
2004-02-15
The apolipoprotein A5 (APOA5) gene has been shown to be important in lowering plasma triglyceride levels. Since several studies have shown that hyperinsulinemia is associated with hypertriglyceridemia, we sought to determine whether APOA5 gene is regulated by insulin. We show here that cell and mouse treatments with insulin down-regulated APOA5 expression in a dose-dependent manner. Furthermore, we determined that insulin decreases APOA5 promoter activity and subsequent deletion analyses revealed an E-box-containing fragment. We showed that Upstream Stimulatory Factors, USF1/USF2, bind to the identified E-box in the APOA5 promoter. Moreover, in cotransfection studies, USF1 stimulates APOA5 promoter activity. The treatment withmore » insulin reduces the binding of USF1/USF2 to APOA5 promoter. The inhibition of PI3K pathway with wortmannin abolished the insulin s effect on APOA5 gene transcription. Using oligoprecipitation method of USF from nuclear extracts, we demonstrated that phosphorylated USF1 failed to bind to APOA5 promoter. This indicates that the APOA5 gene transrepression by insulin involves a phosphorylation of USF through PI3K, that modulate their binding to APOA5 promoter and results in APOA5 down-regulation. The effect of exogenous hyperinsulinemia in healthy men shows a decrease of the plasma ApoAV level. These data suggest a potential mechanism involving APOA5 gene in hypertriglyceridemia associated with hyperinsulinemia.« less
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Dahabieh, Matthew S., E-mail: dahabieh@interchange.ubc.ca; Ooms, Marcel, E-mail: marcel.ooms@mssm.edu; Malcolm, Tom, E-mail: tmalc1@yahoo.com
Transcription from the HIV-1 long terminal repeat (LTR) is mediated by numerous host transcription factors. In this study we characterized an E-box motif (RBE1) within the core promoter that was previously implicated in both transcriptional activation and repression. We show that RBE1 is a binding site for the RBF-2 transcription factor complex (USF1, USF2, and TFII-I), previously shown to bind an upstream viral element, RBE3. The RBE1 and RBE3 elements formed complexes of identical mobility and protein constituents in gel shift assays, both with Jurkat T-cell nuclear extracts and recombinant USF/TFII-I. Furthermore, both elements are regulators of HIV-1 expression; mutationsmore » in LTR-luciferase reporters and in HIV-1 molecular clones resulted in decreased transcription, virion production, and proviral expression in infected cells. Collectively, our data indicate that RBE1 is a bona fide RBF-2 binding site and that the RBE1 and RBE3 elements are necessary for mediating proper transcription from the HIV-1 LTR.« less
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Khund-Sayeed, Syed; He, Ximiao; Holzberg, Timothy; Wang, Jun; Rajagopal, Divya; Upadhyay, Shriyash; Durell, Stewart R; Mukherjee, Sanjit; Weirauch, Matthew T; Rose, Robert; Vinson, Charles
2016-09-12
We evaluated DNA binding of the B-HLH family members TCF4 and USF1 using protein binding microarrays (PBMs) containing double-stranded DNA probes with cytosine on both strands or 5-methylcytosine (5mC) or 5-hydroxymethylcytosine (5hmC) on one DNA strand and cytosine on the second strand. TCF4 preferentially bound the E-box motif (CAN|NTG) with strongest binding to the 8-mer CAG|GTGGT. 5mC uniformly decreases DNA binding of both TCF4 and USF1. The bulkier 5hmC also inhibited USF1 binding to DNA. In contrast, 5hmC dramatically enhanced TCF4 binding to E-box motifs ACAT|GTG and ACAC|GTG, being better bound than any 8-mer containing cytosine. Examination of X-ray structures of the closely related TCF3 and USF1 bound to DNA suggests TCF3 can undergo a conformational shift to preferentially bind to 5hmC while the USF1 basic region is bulkier and rigid precluding a conformation shift to bind 5hmC. These results greatly expand the regulatory DNA sequence landscape bound by TCF4.
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Qiao, Huan; May, James M.
2013-01-01
SVCT2 is the major transporter mediating vitamin C uptake in most organs. Its expression is driven by two promoters (CpG-poor exon 1a promoter and CpG-rich exon 1b promoter). In this work we mapped discrete elements within the proximal CpG-poor promoter responsible for the exon 1a transcription. We identified two E boxes for USF binding and one Y box for NF-Y binding. We further show that the formation of an NFY/USF complex on the exon 1a promoter amplifies each other's ability to bind to the promoter in a cooperativity-dependent manner and is absolutely required for the full activity of the exon 1a promoter. The analysis of the CpG site located at the upstream USF binding site in the promoter showed a strong correlation between expression and demethylation. It was also shown that the exon 1a transcription was induced in cell culture treated with demethylating agent decitabine. The specific methylation of this CpG site impaired both the binding of USF and the formation of the functional NF-Y/USF complex as well as promoter activity, suggesting its importance for the cell-specific transcription. Thus CpG methylation at the upstream USF binding site functions in establishing and maintaining cell-specific transcription from the CpG-poor SVCT2 exon 1a promoter. PMID:21770893
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Maekawa, T; Sudo, T; Kurimoto, M; Ishii, S
1991-09-11
The transcription factor HIV-TF1, which binds to a region about 60 bp upstream from the enhancer of the human immunodeficiency virus-1 (HIV-1), was purified from human B cells. HIV-TF1 had a molecular weight of 39,000. Binding of HIV-TF1 to the HIV long terminal repeat (LTR) activated transcription from the HIV promoter in vitro. The HIV-TF1-binding site in HIV LTR was similar to the site recognized by upstream stimulatory factor (USF) in the adenovirus major late promoter. DNA-binding properties of HIV-TF1 suggested that HIV-TF1 might be identical or related to USF. Interestingly, treatment of purified HIV-TF1 by phosphatase greatly reduced its DNA-binding activity, suggesting that phosphorylation of HIV-TF1 was essential for DNA binding. The disruption of HIV-TF1-binding site induced a 60% decrease in the level of transcription from the HIV promoter in vivo. These results suggest that HIV-TF1 is involved in transcriptional regulation of HIV-1.
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STAT3 or USF2 Contributes to HIF Target Gene Specificity
Pawlus, Matthew R.; Wang, Liyi; Murakami, Aya; Dai, Guanhai; Hu, Cheng-Jun
2013-01-01
The HIF1- and HIF2-mediated transcriptional responses play critical roles in solid tumor progression. Despite significant similarities, including their binding to promoters of both HIF1 and HIF2 target genes, HIF1 and HIF2 proteins activate unique subsets of target genes under hypoxia. The mechanism for HIF target gene specificity has remained unclear. Using siRNA or inhibitor, we previously reported that STAT3 or USF2 is specifically required for activation of endogenous HIF1 or HIF2 target genes. In this study, using reporter gene assays and chromatin immuno-precipitation, we find that STAT3 or USF2 exhibits specific binding to the promoters of HIF1 or HIF2 target genes respectively even when over-expressed. Functionally, HIF1α interacts with STAT3 to activate HIF1 target gene promoters in a HIF1α HLH/PAS and N-TAD dependent manner while HIF2α interacts with USF2 to activate HIF2 target gene promoters in a HIF2α N-TAD dependent manner. Physically, HIF1α HLH and PAS domains are required for its interaction with STAT3 while both N- and C-TADs of HIF2α are involved in physical interaction with USF2. Importantly, addition of functional USF2 binding sites into a HIF1 target gene promoter increases the basal activity of the promoter as well as its response to HIF2+USF2 activation while replacing HIF binding site with HBS from a HIF2 target gene does not change the specificity of the reporter gene. Importantly, RNA Pol II on HIF1 or HIF2 target genes is primarily associated with HIF1α or HIF2α in a STAT3 or USF2 dependent manner. Thus, we demonstrate here for the first time that HIF target gene specificity is achieved by HIF transcription partners that are required for HIF target gene activation, exhibit specific binding to the promoters of HIF1 or HIF2 target genes and selectively interact with HIF1α or HIF2α protein. PMID:23991099
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Vallejo, Griselda; Mestre-Citrinovitz, Ana C.; Ballaré, Cecilia; Beato, Miguel; Saragüeta, Patricia
2014-01-01
Although non-genomic steroid receptor pathways have been studied over the past decade, little is known about the direct gene expression changes that take place as a consequence of their activation. Progesterone controls proliferation of rat endometrial stromal cells during the peri-implantation phase of pregnancy. We showed that picomolar concentration of progestin R5020 mimics this control in UIII endometrial stromal cells via ERK1-2 and AKT activation mediated by interaction of Progesterone Receptor (PR) with Estrogen Receptor beta (ERb) and without transcriptional activity of endogenous PR and ER. Here we identify early downstream targets of cytoplasmic PR signaling and their possible role in endometrial stromal cell proliferation. Microarray analysis of global gene expression changes in UIII cells treated for 45 min with progestin identified 97 up- and 341 down-regulated genes. The most over-represented molecular functions were transcription factors and regulatory factors associated with cell proliferation and cell cycle, a large fraction of which were repressors down-regulated by hormone. Further analysis verified that progestins regulate Ccnd1, JunD, Usf1, Gfi1, Cyr61, and Cdkn1b through PR-mediated activation of ligand-free ER, ERK1-2 or AKT, in the absence of genomic PR binding. ChIP experiments show that progestin promoted the interaction of USF1 with the proximal promoter of the Cdc2 gene. Usf1 knockdown abolished Cdc2 progestin-dependent transcriptional regulation and cell proliferation, which also blocked Cdc2 knockdown. We conclude that progestin-induced proliferation of endometrial stromal cells is mediated by ERK1-2 and AKT dependent early regulation of USF1, which directly induces Cdc2. To our knowledge, this is the first description of early target genes of progestin-activated classical PR via crosstalk with protein kinases and independently of hormone receptor binding to the genomic targets. PMID:24859236
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The transcription factor DREAM represses A20 and mediates inflammation
Tiruppathi, Chinnaswamy; Soni, Dheeraj; Wang, Dong-Mei; Xue, Jiaping; Singh, Vandana; Thippegowda, Prabhakar B.; Cheppudira, Bopaiah P.; Mishra, Rakesh K.; DebRoy, Auditi; Qian, Zhijian; Bachmaier, Kurt; Zhao, Youyang; Christman, John W.; Vogel, Stephen M.; Ma, Averil; Malik, Asrar B.
2014-01-01
Here we show that the transcription-repressor DREAM binds to the A20 promoter to repress the expression of A20, the deubiquitinase suppressing inflammatory NF-κB signaling. DREAM-deficient (Dream−/−) mice displayed persistent and unchecked A20 expression in response to endotoxin. DREAM functioned by transcriptionally repressing A20 through binding to downstream regulatory elements (DREs). In contrast, USF1 binding to the DRE-associated E-box domain activated A20 expression in response to inflammatory stimuli. These studies define the critical opposing functions of DREAM and USF1 in inhibiting and inducing A20 expression, respectively, and thereby the strength of NF-κB signaling. Targeting of DREAM to induce USF1-mediated A20 expression is therefore a potential anti-inflammatory strategy in diseases such as acute lung injury associated with unconstrained NF-κB activity. PMID:24487321
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Qiao, Huan; May, James M.
2012-01-01
Transcription of the ascorbate transporter, SVCT2, is driven by two distinct promoters in exon 1 of the transporter sequence. The exon 1a promoter lacks a classical transcription start site and little is known about regulation of promoter activity in the transcription start site core (TSSC) region. Here we present evidence that the TSSC binds the multifunctional initiator-binding protein YY1. Electrophoresis shift assays using YY1 antibody showed that YY1 is present as one of two major complexes that specifically bind to the TSSC. The other complex contains the transcription factor NF-Y. Mutations in the TSSC that decreased YY1 binding also impaired the exon 1a promoter activity despite the presence of an upstream activating NF-Y/USF complex, suggesting that YY1 is involved in the regulation of the exon 1a transcription. Furthermore, YY1 interaction with NF-Y and/or USF synergistically enhanced the exon 1a promoter activity in transient transfections and co-activator p300 enhanced their synergistic activation. We propose that the TSSC plays a vital role in the exon 1a transcription and that this function is partially carried out by the transcription factor YY1. Moreover, co-activator p300 might be able to synergistically enhance the TSSC function via a “bridge” mechanism with upstream sequences. PMID:22532872
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Glucose Regulates the Expression of the Apolipoprotein A5 Gene
DOE Office of Scientific and Technical Information (OSTI.GOV)
Fruchart, Jamila; Nowak, Maxime; Helleboid-Chapman, Audrey
2008-04-07
The apolipoprotein A5 gene (APOA5) is a key player in determining triglyceride concentrations in humans and mice. Since diabetes is often associated with hypertriglyceridemia, this study explores whether APOA5 gene expression is regulated by alteration in glucose homeostasis and the related pathways. D-glucose activates APOA5 gene expression in a time- and dose-dependent manner in hepatocytes, and the glycolytic pathway involved was determined using D-glucose analogs and metabolites. Together, transient transfections, electrophoretic mobility shift assays and chromatin immunoprecipitation assays show that this regulation occurs at the transcriptional level through an increase of USF1/2 binding to an E-box in the APOA5 promoter.more » We show that this phenomenon is not due to an increase of mRNA or protein expression levels of USF. Using protein phosphatases 1 and 2A inhibitor, we demonstrate that D-glucose regulates APOA5 gene via a dephosphorylation mechanism, thereby resulting in an enhanced USF1/2-promoter binding. Last, subsequent suppressions of USF1/2 and phosphatases mRNA through siRNA gene silencing abolished the regulation. We demonstrate that APOA5 gene is up regulated by D-glucose and USF through phosphatase activation. These findings may provide a new cross talk between glucose and lipid metabolism.« less
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Zeng, Yanli; Li, Hui; Zhang, Xiaoju
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (APOBEC3G, A3G) exert antiviral defense as an important factor of innate immunity. A variety of cytokines such as IFN-γ,IL2,IL15,IL7 could induce the transcription of A3G. However, the regulation of other nuclear factor on the transcription of A3G have not been reported at the present. To gain new insights into the transcriptional regulation of this restriction factor, we cloned and characterized the promoter region of A3G and investigate the modulation of USF1 gene on the transcription of A3G. We identified a 232 bp region that was sufficient to regulate the activity of full promoter. Transcriptionalmore » start sites (TSS) were identified by the luciferase reporter assays of plasmids containing full or shorter fragments of the A3G promoter. The results demonstrated that the core promoter of A3G is located within the region -159/-84 relative to the TSS. Transcriptional activity of A3G core promoter regulated by USF1 was dependent on an E-box (located at position -91/-86 relative to the major TSS) and was abolished after mutation of this DNA element. USF1 gene can take part in basal transcription regulation of the human A3G gene in hepatocyte, and the identified E-box represented a binding site for the USF1. - Highlights: • The core promoter of A3G is located within the region −159/−84 relative to the TSS. • Transcriptional activity of A3G core promoter regulated by USF1 was dependent on an E-box (located at position −91/−86 relative to the major TSS). • USF1 gene can take part in basal transcription regulation of the human A3G gene in hepatocyte.« less
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Insulin signalling mechanisms for triacylglycerol storage.
Czech, M P; Tencerova, M; Pedersen, D J; Aouadi, M
2013-05-01
Insulin signalling is uniquely required for storing energy as fat in humans. While de novo synthesis of fatty acids and triacylglycerol occurs mostly in liver, adipose tissue is the primary site for triacylglycerol storage. Insulin signalling mechanisms in adipose tissue that stimulate hydrolysis of circulating triacylglycerol, uptake of the released fatty acids and their conversion to triacylglycerol are poorly understood. New findings include (1) activation of DNA-dependent protein kinase to stimulate upstream stimulatory factor (USF)1/USF2 heterodimers, enhancing the lipogenic transcription factor sterol regulatory element binding protein 1c (SREBP1c); (2) stimulation of fatty acid synthase through AMP kinase modulation; (3) mobilisation of lipid droplet proteins to promote retention of triacylglycerol; and (4) upregulation of a novel carbohydrate response element binding protein β isoform that potently stimulates transcription of lipogenic enzymes. Additionally, insulin signalling through mammalian target of rapamycin to activate transcription and processing of SREBP1c described in liver may apply to adipose tissue. Paradoxically, insulin resistance in obesity and type 2 diabetes is associated with increased triacylglycerol synthesis in liver, while it is decreased in adipose tissue. This and other mysteries about insulin signalling and insulin resistance in adipose tissue make this topic especially fertile for future research.
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Cruickshank, Mark N; Dods, James; Taylor, Rhonda L; Karimi, Mahdad; Fenwick, Emily J; Quail, Elizabeth A; Rea, Alexander J; Holers, V Michael; Abraham, Lawrence J; Ulgiati, Daniela
2015-07-01
Complement receptor 2 (CR2/CD21) plays an important role in the generation of normal B cell immune responses. As transcription appears to be the prime mechanism via which surface CR2/CD21 expression is controlled, understanding transcriptional regulation of this gene will have broader implications to B cell biology. Here we report opposing, cell-context specific control of CR2/CD21 promoter activity by tandem E-box elements, spaced 22 bp apart and within 70 bp of the transcription initiation site. We have identified E2A and USF transcription factors as binding to the distal and proximal E-box sites respectively in CR2-positive B-cells, at a site that is hypersensitive to restriction enzyme digestion compared to non-expressing K562 cells. However, additional unidentified proteins have also been found to bind these functionally important elements. By utilizing a proteomics approach we have identified a repressor protein, RP58, binding the distal E-box motif. Co-transfection experiments using RP58 overexpression constructs demonstrated a specific 10-fold repression of CR2/CD21 transcriptional activity mediated through the distal E-box repressor element. Taken together, our results indicate that repression of the CR2/CD21 promoter can occur through one of the E-box motifs via recruitment of RP58 and other factors to bring about a silenced chromatin context within CR2/CD21 non-expressing cells. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Köberle, Martin; Göppel, David; Grandl, Tanja; Gaentzsch, Peer; Manncke, Birgit; Berchtold, Susanne; Müller, Steffen; Lüscher, Bernhard; Asselin-Labat, Marie-Liesse; Pallardy, Marc; Sorg, Isabel; Langer, Simon; Barth, Holger; Zumbihl, Robert; Autenrieth, Ingo B.; Bohn, Erwin
2012-01-01
Glucocorticoid induced-leucine zipper (GILZ) has been shown to be induced in cells by different stimuli such as glucocorticoids, IL-10 or deprivation of IL-2. GILZ has anti-inflammatory properties and may be involved in signalling modulating apoptosis. Herein we demonstrate that wildtype Yersinia enterocolitica which carry the pYV plasmid upregulated GILZ mRNA levels and protein expression in epithelial cells. Infection of HeLa cells with different Yersinia mutant strains revealed that the protease activity of YopT, which cleaves the membrane-bound form of Rho GTPases was sufficient to induce GILZ expression. Similarly, Clostridium difficile toxin B, another bacterial inhibitor of Rho GTPases induced GILZ expression. YopT and toxin B both increased transcriptional activity of the GILZ promoter in HeLa cells. GILZ expression could not be linked to the inactivation of an individual Rho GTPase by these toxins. However, forced expression of RhoA and RhoB decreased basal GILZ promoter activity. Furthermore, MAPK activation proved necessary for profound GILZ induction by toxin B. Promoter studies and gel shift analyses defined binding of upstream stimulatory factor (USF) 1 and 2 to a canonical c-Myc binding site (E-box) in the GILZ promoter as a crucial step of its trans-activation. In addition we could show that USF-1 and USF-2 are essential for basal as well as toxin B induced GILZ expression. These findings define a novel way of GILZ promoter trans-activation mediated by bacterial toxins and differentiate it from those mediated by dexamethasone or deprivation of IL-2. PMID:22792400
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Carmi-Levy, Irit; Yannay-Cohen, Nurit; Kay, Gillian; Razin, Ehud; Nechushtan, Hovav
2008-01-01
We previously discovered that microphthalmia transcription factor (MITF) and upstream stimulatory factor 2 (USF2) each forms a complex with its inhibitor histidine triad nucleotide-binding 1 (Hint-1) and with lysyl-tRNA synthetase (LysRS). Moreover, we showed that the dinucleotide diadenosine tetraphosphate (Ap4A), previously shown to be synthesized by LysRS, binds to Hint-1, and as a result the transcription factors are released from their suppression. Thus, transcriptional activity is regulated by Ap4A, suggesting that Ap4A is a second messenger in this context. For Ap4A to be unambiguously established as a second messenger, several criteria have to be fulfilled, including the presence of a metabolizing enzyme. Since several enzymes are able to hydrolize Ap4A, we provided here evidence that the “Nudix” type 2 gene product, Ap4A hydrolase, is responsible for Ap4A degradation following the immunological activation of mast cells. The knockdown of Ap4A hydrolase modulated Ap4A accumulation, resulting in changes in the expression of MITF and USF2 target genes. Moreover, our observations demonstrated that the involvement of Ap4A hydrolase in gene regulation is not a phenomenon exclusive to mast cells but can also be found in cardiac cells activated with the β-agonist isoproterenol. Thus, we have provided concrete evidence establishing Ap4A as a second messenger in the regulation of gene expression. PMID:18644867
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Carmi-Levy, Irit; Yannay-Cohen, Nurit; Kay, Gillian; Razin, Ehud; Nechushtan, Hovav
2008-09-01
We previously discovered that microphthalmia transcription factor (MITF) and upstream stimulatory factor 2 (USF2) each forms a complex with its inhibitor histidine triad nucleotide-binding 1 (Hint-1) and with lysyl-tRNA synthetase (LysRS). Moreover, we showed that the dinucleotide diadenosine tetraphosphate (Ap(4)A), previously shown to be synthesized by LysRS, binds to Hint-1, and as a result the transcription factors are released from their suppression. Thus, transcriptional activity is regulated by Ap(4)A, suggesting that Ap(4)A is a second messenger in this context. For Ap(4)A to be unambiguously established as a second messenger, several criteria have to be fulfilled, including the presence of a metabolizing enzyme. Since several enzymes are able to hydrolyze Ap(4)A, we provided here evidence that the "Nudix" type 2 gene product, Ap(4)A hydrolase, is responsible for Ap(4)A degradation following the immunological activation of mast cells. The knockdown of Ap(4)A hydrolase modulated Ap(4)A accumulation, resulting in changes in the expression of MITF and USF2 target genes. Moreover, our observations demonstrated that the involvement of Ap(4)A hydrolase in gene regulation is not a phenomenon exclusive to mast cells but can also be found in cardiac cells activated with the beta-agonist isoproterenol. Thus, we have provided concrete evidence establishing Ap(4)A as a second messenger in the regulation of gene expression.
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Mxi1 is a repressor of the c-Myc promoter and reverses activation by USF.
Lee, T C; Ziff, E B
1999-01-08
The basic region/helix-loop-helix/leucine zipper (B-HLH-LZ) oncoprotein c-Myc is abundant in proliferating cells and forms heterodimers with Max protein that bind to E-box sites in DNA and stimulate genes required for proliferation. A second B-HLH-LZ protein, Mxi1, is induced during terminal differentiation, and forms heterodimers with Max that also bind E-boxes but tether the mSin3 transcriptional repressor protein along with histone deacetylase thereby antagonizing Myc-dependent activation. We show that Mxi1 also antagonizes Myc by a second pathway, repression of transcription from the major c-myc promoter, P2. Repression was independent of Mxi1 binding to mSin3 but dependent on the Mxi1 LZ and COOH-terminal sequences, including putative casein kinase II phosphorylation sites. Repression targeted elements of the myc P2 promoter core (-35/+10), where it reversed transactivation by the constitutive transcription factor, USF. We show that Zn2+ induction of a stably transfected, metallothionein promoter-regulated mxi1 gene blocked the ability of serum to induce transcription of the endogenous c-myc gene and cell entry into S phase. Thus, induction of Mxi1 in terminally differentiating cells may block Myc function by repressing the c-myc gene P2 promoter, as well as by antagonizing Myc-dependent transactivation through E-boxes.
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Nayak, G; Cooper, G M
2012-10-11
The phosphatidylinositol (PI) 3-kinase/Akt signaling pathway has a prominent role in cell survival and proliferation, in part, by regulating gene expression at the transcriptional level. Previous work using global expression profiling identified FOXOs and the E-box-binding transcription factors MITF and USF1 as key targets of PI 3-kinase signaling that lead to the induction of proapoptotic and cell cycle arrest genes in response to inhibition of PI 3-kinase. In this study, we investigated the role of p53 downstream of PI 3-kinase signaling by analyzing the effects of inhibition of PI 3-kinase in Rat-1 cells, which have wild-type p53, compared with Rat-1 cells expressing a dominant-negative p53 mutant. Expression of dominant-negative p53 conferred partial resistance to apoptosis induced by inhibition of PI 3-kinase. Global gene expression profiling combined with computational and experimental analysis of transcription factor binding sites demonstrated that p53, along with FOXO, MITF and USF1, contributed to gene induction in response to PI 3-kinase inhibition. Activation of p53 was mediated by phosphorylation of the histone acetyltransferase Tip60 by glycogen synthase kinase (GSK) 3, leading to activation of p53 by acetylation. Many of the genes targeted by p53 were also targeted by FOXO and E-box-binding transcription factors, indicating that p53 functions coordinately with these factors to regulate gene expression downstream of PI 3-kinase/Akt/GSK3 signaling.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Adam, Tasneem; Opie, Lionel H.; Essop, M. Faadiel, E-mail: mfessop@sun.ac.za
Research highlights: {yields} AMPK inhibits acetyl-CoA carboxylase beta gene promoter activity. {yields} Nuclear respiratory factor-1 inhibits acetyl-CoA carboxylase beta promoter activity. {yields} AMPK regulates acetyl-CoA carboxylase beta at transcriptional level. -- Abstract: The cardiac-enriched isoform of acetyl-CoA carboxylase (ACC{beta}) produces malonyl-CoA, a potent inhibitor of carnitine palmitoyltransferase-1. AMPK inhibits ACC{beta} activity, lowering malonyl-CoA levels and promoting mitochondrial fatty acid {beta}-oxidation. Previously, AMPK increased promoter binding of nuclear respiratory factor-1 (NRF-1), a pivotal transcriptional modulator controlling gene expression of mitochondrial proteins. We therefore hypothesized that NRF-1 inhibits myocardial ACC{beta} promoter activity via AMPK activation. A human ACC{beta} promoter-luciferase construct was transientlymore » transfected into neonatal cardiomyocytes {+-} a NRF-1 expression construct. NRF-1 overexpression decreased ACC{beta} gene promoter activity by 71 {+-} 4.6% (p < 0.001 vs. control). Transfections with 5'-end serial promoter deletions revealed that NRF-1-mediated repression of ACC{beta} was abolished with a pPII{beta}-18/+65-Luc deletion construct. AMPK activation dose-dependently reduced ACC{beta} promoter activity, while NRF-1 addition did not further decrease it. We also investigated NRF-1 inhibition in the presence of upstream stimulatory factor 1 (USF1), a known transactivator of the human ACC{beta} gene promoter. Here NRF-1 blunted USF1-dependent induction of ACC{beta} promoter activity by 58 {+-} 7.5% (p < 0.001 vs. control), reversed with a dominant negative NRF-1 construct. NRF-1 also suppressed endogenous USF1 transcriptional activity by 55 {+-} 6.2% (p < 0.001 vs. control). This study demonstrates that NRF-1 is a novel transcriptional inhibitor of the human ACC{beta} gene promoter in the mammalian heart. Our data extends AMPK regulation of ACC{beta} to the transcriptional level.« less
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Legraverend, C; Antonson, P; Flodby, P; Xanthopoulos, K G
1993-01-01
The promoter region of the mouse CCAAT-Enhancer Binding Protein (C/EBP alpha) gene is capable of directing high levels of expression of reporter constructs in various cell lines, albeit even in cells that do not express their endogenous C/EBP alpha gene. To understand the molecular mechanisms underlying this ubiquitous expression, we have characterized the promoter region of the mouse C/EBP alpha gene by a variety of in vitro and in vivo methods. We show that three sites related in sequence to USF, BTE and C/EBP binding sites and present in promoter region -350/+3, are recognized by proteins from rat liver nuclear extracts. The sequence of the C/EBP alpha promoter that includes the USF binding site is also capable of forming stable complexes with purified Myc+Max heterodimers and mutation of this site drastically reduces transcription of C/EBP alpha promoter luciferase constructs both in liver and non liver cell lines. In addition, we identify three novel protein-binding sites two of which display similarity to NF-1 and a NF kappa B binding sites. The region located between nucleotides -197 and -178 forms several heat-stable complexes with liver nuclear proteins in vitro which are recognized mainly by antibodies specific for C/EBP alpha. Furthermore, transient expression of C/EBP alpha and to a lesser extent C/EBP beta expression vectors, results in transactivation of a cotransfected C/EBP alpha promoter-luciferase reporter construct. These experiments support the notion that the C/EBP alpha gene is regulated by C/EBP alpha but other C/EBP-related proteins may also be involved. Images PMID:8493090
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Niemiec, Pawel; Nowak, Tomasz; Iwanicki, Tomasz; Gorczynska-Kosiorz, Sylwia; Balcerzyk, Anna; Krauze, Jolanta; Grzeszczak, Wladyslaw; Wiecha, Maria; Zak, Iwona
2015-01-01
Single nucleotide polymorphisms (SNPs) of the USF1 gene (upstream stimulatory factor 1) influence plasma lipid levels. This study aims to determine whether USF1 SNPs interact with traditional risk factors of atherosclerosis to increase coronary artery disease (CAD) risk. In the present study serum lipid levels and USF1 gene polymorphisms (rs2516839 and rs3737787) were determined in 470 subjects: 235 patients with premature CAD and 235 controls. A trend of increasing triglycerides (TG) levels in relation to the C allele dose of rs2516839 SNP was observed. The synergistic effect of cigarette smoking and C allele carrier state on CAD risk was also found (SIM = 2.69, p = 0.015). TG levels differentiated significantly particular genotypes in smokers (1.53 mmol/L for TT, 1.80 mmol/L for CT and 2.27 mmol/L for CC subjects). In contrast, these differences were not observed in the non-smokers subgroup (1.57 mmol/L for TT, 1.46 mmol/L for CT and 1.49 mmol/L for CC subjects). In conclusion, the rs2516839 polymorphism may modulate serum triglyceride levels in response to cigarette smoking. Carriers of the C allele seem to be particularly at risk of CAD, when exposed to cigarette smoking. PMID:26068452
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Kirpich, Irina A; Feng, Wenke; Wang, Yuhua; Liu, Yanlong; Beier, Juliane I; Arteel, Gavin E; Falkner, K Cameron; Barve, Shirish S; McClain, Craig J
2013-05-01
Alcohol and dietary fat both play an important role in alcohol-mediated multi-organ pathology, including gut and liver. In the present study we hypothesized that the combination of alcohol and dietary unsaturated fat (USF) would result in intestinal inflammatory stress and mucus layer alterations, thus contributing to disruption of intestinal barrier integrity. C57BL/6N mice were fed Lieber-DeCarli liquid diets containing EtOH and enriched in USF (corn oil/linoleic acid) or SF (medium chain triglycerides: beef tallow) for 8 weeks. Intestinal histology, morphometry, markers of inflammation, as well as levels of mucus protective factors were evaluated. Alcohol and dietary USF triggered an intestinal pro-inflammatory response, characterized by increase in Tnf-α, MCP1, and MPO activity. Further, alcohol and dietary USF, but not SF, resulted in alterations of the intestinal mucus layer, characterized by decreased expression of Muc2 in the ileum. A strong correlation was observed between down-regulation of the antimicrobial factor Cramp and increased Tnf-α mRNA. Therefore, dietary unsaturated fat (corn oil/LA enriched) is a significant contributing factor to EtOH-mediated intestinal inflammatory response and mucus layer alterations in rodents. Copyright © 2013 Elsevier Inc. All rights reserved.
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2016-01-01
Color variation provides the opportunity to investigate the genetic basis of evolution and selection. Reptiles are less studied than mammals. Comparative genomics approaches allow for knowledge gained in one species to be leveraged for use in another species. We describe a comparative vertebrate analysis of conserved regulatory modules in pythons aimed at assessing bioinformatics evidence that transcription factors important in mammalian pigmentation phenotypes may also be important in python pigmentation phenotypes. We identified 23 python orthologs of mammalian genes associated with variation in coat color phenotypes for which we assessed the extent of pairwise protein sequence identity between pythons and mouse, dog, horse, cow, chicken, anole lizard, and garter snake. We next identified a set of melanocyte/pigment associated transcription factors (CREB, FOXD3, LEF-1, MITF, POU3F2, and USF-1) that exhibit relatively conserved sequence similarity within their DNA binding regions across species based on orthologous alignments across multiple species. Finally, we identified 27 evolutionarily conserved clusters of transcription factor binding sites within ~200-nucleotide intervals of the 1500-nucleotide upstream regions of AIM1, DCT, MC1R, MITF, MLANA, OA1, PMEL, RAB27A, and TYR from Python bivittatus. Our results provide insight into pigment phenotypes in pythons. PMID:27698666
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Irizarry, Kristopher J L; Bryden, Randall L
2016-01-01
Color variation provides the opportunity to investigate the genetic basis of evolution and selection. Reptiles are less studied than mammals. Comparative genomics approaches allow for knowledge gained in one species to be leveraged for use in another species. We describe a comparative vertebrate analysis of conserved regulatory modules in pythons aimed at assessing bioinformatics evidence that transcription factors important in mammalian pigmentation phenotypes may also be important in python pigmentation phenotypes. We identified 23 python orthologs of mammalian genes associated with variation in coat color phenotypes for which we assessed the extent of pairwise protein sequence identity between pythons and mouse, dog, horse, cow, chicken, anole lizard, and garter snake. We next identified a set of melanocyte/pigment associated transcription factors (CREB, FOXD3, LEF-1, MITF, POU3F2, and USF-1) that exhibit relatively conserved sequence similarity within their DNA binding regions across species based on orthologous alignments across multiple species. Finally, we identified 27 evolutionarily conserved clusters of transcription factor binding sites within ~200-nucleotide intervals of the 1500-nucleotide upstream regions of AIM1, DCT, MC1R, MITF, MLANA, OA1, PMEL, RAB27A, and TYR from Python bivittatus . Our results provide insight into pigment phenotypes in pythons.
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Zhang, Leying; Handel, Michelle Van; Schartner, Jill M; Hagar, Aaron; Allen, Grant; Curet, Marjorie; Badie, Behnam
2007-03-01
Understanding the local CNS immune response to neoplasms is essential in the development of immune-based treatments for malignant brain tumors. Using rodent glioma models, we have recently found tumor-associated microglia/macrophages (MG/MP) to be less responsive to known MG/MP activators such as CpG, LPS and IFN-gamma. To understand the mechanism of MG/MP suppression, nuclear extracts from rodent intracranial C6 gliomas, C6 glioma-associated MG/MP, normal brain, and normal MG/MP were obtained and studied using Electrophoretic Mobility Shift Assay (EMSA). Among the nuclear factors studied (AP-1, IRF, USF-1 and Stat-1) only USF-1, which is constitutively expressed in most cells, was down-regulated in tumor-associated MG/MP, but not normal MG/MP. Because tumor-associated MG/MP had higher expression of IL-10 (but not TNF-alpha or TGF-beta), we evaluated the role of USF-1 on IL-10 expression. siRNA mediated inhibition of USF-1 expression in primary MG/MP cultures resulted in up-regulation of IL-10 mRNA but not TNF-alpha or TGF-beta. These findings suggest that USF-1 may play a role in IL-10 regulation in MG/MP in brain tumors.
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Transcriptional regulation of hepatic lipogenesis.
Wang, Yuhui; Viscarra, Jose; Kim, Sun-Joong; Sul, Hei Sook
2015-11-01
Fatty acid and fat synthesis in the liver is a highly regulated metabolic pathway that is important for very low-density lipoprotein (VLDL) production and thus energy distribution to other tissues. Having common features at their promoter regions, lipogenic genes are coordinately regulated at the transcriptional level. Transcription factors, such as upstream stimulatory factors (USFs), sterol regulatory element-binding protein 1C (SREBP1C), liver X receptors (LXRs) and carbohydrate-responsive element-binding protein (ChREBP) have crucial roles in this process. Recently, insights have been gained into the signalling pathways that regulate these transcription factors. After feeding, high blood glucose and insulin levels activate lipogenic genes through several pathways, including the DNA-dependent protein kinase (DNA-PK), atypical protein kinase C (aPKC) and AKT-mTOR pathways. These pathways control the post-translational modifications of transcription factors and co-regulators, such as phosphorylation, acetylation or ubiquitylation, that affect their function, stability and/or localization. Dysregulation of lipogenesis can contribute to hepatosteatosis, which is associated with obesity and insulin resistance.
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Oh, Mi Mi; Kim, Jin Wook; Park, Min Gu; Kim, Je Jong; Yoo, Kee Hwan; Moon, Du Geon
2012-03-01
We assessed the role of therapeutic delay time (TDT) in acute renal cortical scintigraphic lesion (ASL) and ultimate scar formation (USF) in children with first febrile UTI and whether it is affected by the presence of vesico-ureteral reflux (VUR). 230 children, 90 girls and 140 boys with first febrile UTI were included. Radiologic (USG, DMSA, and VCUG), clinical (age, gender, peak fever, therapeutic delay time) and laboratory (CBC with differential count, ANC (absolute neutrophil count), BUN, Creatinine, urine analysis, gram stain, culture, CRP and ESR) variables were analysed. DMSA was performed within 5 days and after six months. VCUG was performed after acute phase of UTI. The differences in TDT according to the presence of ASL, USF and VUR were assessed. And the correlation between ASL or USF with the duration of TDT was assessed. Of 230 patients enrolled, 142 patients had refluxing UTI and 88 patients had non-refluxing UTI. TDT was the risk factor associated with ASL and USF along with presence of VUR. TDT was longer in ASL positive group compared with the ASL negative group. Also USF group showed longer TDT compared with those without USF in both refluxing UTI and non refluxing UTI. The TDT was significantly shorter in USF group with the presence of VUR. Positive linear association was noted between prevalence of ASL and USF and duration of TDT. In conclusion, the impact of UTI on formation of USF may be enhanced by the presence of VUR with shorter duration of TDT.
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Lee, Dong-Hoon; Singh, Purnima; Tsark, Walter M. K.; Szabó, Piroska E.
2010-01-01
Background The H19/Igf2 imprinting control region (ICR) functions as an insulator exclusively in the unmethylated maternal allele, where enhancer-blocking by CTCF protein prevents the interaction between the Igf2 promoter and the distant enhancers. DNA methylation inhibits CTCF binding in the paternal ICR allele. Two copies of the chicken β-globin insulator (ChβGI)2 are capable of substituting for the enhancer blocking function of the ICR. Insulation, however, now also occurs upon paternal inheritance, because unlike the H19 ICR, the (ChβGI)2 does not become methylated in fetal male germ cells. The (ChβGI)2 is a composite insulator, exhibiting enhancer blocking by CTCF and chromatin barrier functions by USF1 and VEZF1. We asked the question whether these barrier proteins protected the (ChβGI)2 sequences from methylation in the male germ line. Methodology/Principal Findings We genetically dissected the ChβGI in the mouse by deleting the binding sites USF1 and VEZF1. The methylation of the mutant versus normal (ChβGI)2 significantly increased from 11% to 32% in perinatal male germ cells, suggesting that the barrier proteins did have a role in protecting the (ChβGI)2 from methylation in the male germ line. Contrary to the H19 ICR, however, the mutant (mChβGI)2 lacked the potential to attain full de novo methylation in the germ line and to maintain methylation in the paternal allele in the soma, where it consequently functioned as a biallelic insulator. Unexpectedly, a stricter enhancer blocking was achieved by CTCF alone than by a combination of the CTCF, USF1 and VEZF1 sites, illustrated by undetectable Igf2 expression upon paternal transmission. Conclusions/Significance In this in vivo model, hypomethylation at the ICR position together with fetal growth retardation mimicked the human Silver-Russell syndrome. Importantly, late fetal/perinatal death occurred arguing that strict biallelic insulation at the H19/Igf2 ICR position is not tolerated in development. PMID:20838620
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Genetic Association and Interaction Analysis of USF1 and APOA5 on Lipid Levels and Atherosclerosis
Laurila, Pirkka-Pekka; Naukkarinen, Jussi; Kristiansson, Kati; Ripatti, Samuli; Kauttu, Tuuli; Silander, Kaisa; Salomaa, Veikko; Perola, Markus; Karhunen, Pekka J.; Barter, Philip J.; Ehnholm, Christian; Peltonen, Leena
2011-01-01
Objective USF1 is a ubiquitous transcription factor governing the expression of numerous genes of lipid and glucose metabolism. APOA5 is a well-established candidate gene regulating triglyceride (TG) levels and has been identified as a downstream target of upstream stimulatory factor. No detailed studies about the effect of APOA5 on atherosclerotic lesion formation have been conducted, nor has its potential interaction with USF1 been examined. Methods and Results We analyzed allelic variants of USF1 and APOA5 in families (n=516) ascertained for atherogenic dyslipidemia and in an autopsy series of middle-aged men (n=300) with precise quantitative measurements of atherosclerotic lesions. The impact of previously associated APOA5 variants on TGs was observed in the dyslipidemic families, and variant rs3135506 was associated with size of fibrotic aortic lesions in the autopsy series. The USF1 variant rs2516839, associated previously with atherosclerotic lesions, showed an effect on TGs in members of the dyslipidemic families with documented coronary artery disease. We provide preliminary evidence of gene-gene interaction between these variants in an autopsy series with a fibrotic lesion area in the abdominal aorta (P=0.0028), with TGs in dyslipidemic coronary artery disease subjects (P=0.03), and with high-density lipoprotein cholesterol (P=0.008) in a large population cohort of coronary artery disease patients (n=1065) in which the interaction for TGs was not replicated. Conclusion Our findings in these unique samples reinforce the roles of APOA5 and USF1 variants on cardiovascular phenotypes and suggest that both genes contribute to lipid levels and aortic atherosclerosis individually and possibly through epistatic effects. PMID:19910639
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Di Taranto, Maria Donata; Staiano, Antonino; D'Agostino, Maria Nicoletta; D'Angelo, Antonietta; Bloise, Elena; Morgante, Alberto; Marotta, Gennaro; Gentile, Marco; Rubba, Paolo; Fortunato, Giuliana
2015-02-01
Familial combined hyperlipidemia (FCH) is a polygenic and multifactorial disease characterized by a variable phenotype showing increased levels of triglycerides and/or cholesterol. The aim of this study was to identify single nucleotides (SNPs) in lipid-related genes associated with FCH. Twenty SNPs in lipid-related genes were studied in 142 control subjects and 165 FCH patients after excluding patients with mutations in the LDLR gene and patients with the E2/E2 genotype of APOE. In particular, we studied the 9996G > A (rs2073658) and 11235C > T (rs3737787) variants in the Upstream Stimulatory Factor 1 gene (USF1), and the -1131T > C (rs662799) and S19W (rs3135506) variants in the Apolipoprotein A-V gene (APOA5). We found that the frequencies of these variants differed between patients and controls and that are associated with different lipid profiles. At multivariate logistic regression SNP S19W in APOA5 remained significantly associated with FCH independently of age, sex, BMI, cholesterol and triglycerides. Our results show that the USF1 and APOA5 polymorphisms are associated with FCH and that the S19W SNP in the APOA5 gene is associated to the disease independently of total cholesterol, triglycerides and BMI. However, more extensive studies including other SNPs such as rs2516839 in USF1, are required. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Liguori, Renato; Quaranta, Sandro; Di Fiore, Rosanna; Elce, Ausilia; Castaldo, Giuseppe; Amato, Felice
2014-12-01
Plasminogen activator inhibitor-1 (PAI-1) is the major physiological inhibitor of tissue-type plasminogen activator in plasma and the most important regulator of the fibrinolytic pathway. The 4G/5G polymorphism (rs1799889) in the PAI-1 promoter is associated with altered PAI-1 transcription. We have identified a new 4G/5G allele, in which a T is inserted near the 4G tract or replaces a G in the 5G tract, forming a T plus 4G (T4G) region. This new variant was first identified in two women, one had experienced juvenile myocardial infarction, the other repeated miscarriage; both had increased PAI-1 plasma activity. In view of the important influence of this promoter region on PAI-1 protein plasma level, we performed in vitro evaluation of the effects of the T4G variant on the transcription activity of the PAI-1 gene promoter. In silico prediction analysis showed that presence of the T4G allele disrupts the E-Box region upstream of the T4G variant, altering the affinity of the target sequence for E-Box binding factors like upstream stimulatory factor-1 (USF-1). Basal T4G promoter activity was 50% higher compared to 4G and 5G variants, but it was less stimulated by USF-1 overexpression. We also analyzed the effects of IL-1β and IL-6 on the PAI-1 promoter activity of our three constructs and showed that the T4G variant was less affected by IL-1β than the other variants. These findings indicate that the T4G variant may be a novel risk factor for thrombotic events. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Cao, Chunyu; Wu, Hao; Vasilatos, Shauna N; Chandran, Uma; Qin, Ye; Wan, Yong; Oesterreich, Steffi; Davidson, Nancy E; Huang, Yi
2018-04-06
Our recent studies have shown that cross-talk between histone deacetylase 5 (HDAC5) and lysine-specific demethylase 1 (LSD1) facilitates breast cancer progression. In this work, we demonstrated that regulatory activity at -356 to -100 bp promoter element plays a critical role in governing HDAC5 transcription. By using DNA affinity precipitation and mass spectrometry, we identified a group of factors that bind to this element. Among these factors, Upstream Transcription Factor 1 (USF1) was shown to play a critical role in controlling HDAC5 transcription. Through screening a panel of epigenetic modifying drugs, we showed that a natural bioactive HDAC inhibitor, sulforaphane, downregulated HDAC5 transcription by blocking USF1 activity. Sulforaphane facilitated LSD1 ubiquitination and degradation in an HDAC5-dependent manner. A comparative microarray analysis demonstrated a genome wide cooperative effect of HDAC5 and LSD1 on cancer-related gene expression. shRNA knockdown and sulforaphane inhibition of HDAC5/LSD1 exhibited similar effects on expression of HDAC5/LSD1 target genes. We also showed that coordinated cross-talk of HDAC5 and LSD1 is essential for the antitumor efficacy of sulforaphane. Combination treatment with sulforaphane and a potent LSD1 inhibitor resulted in synergistic growth inhibition in breast cancer cells, but not in normal breast epithelial cells. Furthermore, combined therapy with sulforaphane and LSD1 inhibitor exhibited superior inhibitory effect on MDA-MB-231 xenograft tumor growth. Taken together, our work demonstrates that HDAC5-LSD1 axis is an effective drug target for breast cancer. Inhibition of HDAC5-LSD1 axis with sulforaphane blocks breast cancer growth and combined treatment with LSD1 inhibitor improves the therapeutic efficacy of sulforaphane. © 2018 UICC.
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Human immunodeficiency virus type 1 Nef protein inhibits NF-kappa B induction in human T cells.
Niederman, T M; Garcia, J V; Hastings, W R; Luria, S; Ratner, L
1992-01-01
Human immunodeficiency virus type 1 (HIV-1) can establish a persistent and latent infection in CD4+ T lymphocytes (W. C. Greene, N. Engl. J. Med. 324:308-317, 1991; S. M. Schnittman, M. C. Psallidopoulos, H. C. Lane, L. Thompson, M. Baseler, F. Massari, C. H. Fox, N. P. Salzman, and A. S. Fauci, Science 245:305-308, 1989). Production of HIV-1 from latently infected cells requires host cell activation by T-cell mitogens (T. Folks, D. M. Powell, M. M. Lightfoote, S. Benn, M. A. Martin, and A. S. Fauci, Science 231:600-602, 1986; D. Zagury, J. Bernard, R. Leonard, R. Cheynier, M. Feldman, P. S. Sarin, and R. C. Gallo, Science 231:850-853, 1986). This activation is mediated by the host transcription factor NF-kappa B [G. Nabel and D. Baltimore, Nature (London) 326:711-717, 1987]. We report here that the HIV-1-encoded Nef protein inhibits the induction of NF-kappa B DNA-binding activity by T-cell mitogens. However, Nef does not affect the DNA-binding activity of other transcription factors implicated in HIV-1 regulation, including SP-1, USF, URS, and NF-AT. Additionally, Nef inhibits the induction of HIV-1- and interleukin 2-directed gene expression, and the effect on HIV-1 transcription depends on an intact NF-kappa B-binding site. These results indicate that defective recruitment of NF-kappa B may underlie Nef's negative transcriptional effects on the HIV-1 and interleukin 2 promoters. Further evidence suggests that Nef inhibits NF-kappa B induction by interfering with a signal derived from the T-cell receptor complex. Images PMID:1527859
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Catalyst Bed Instability Within the USFE H2O2/JP-8 Rocket Engine
NASA Technical Reports Server (NTRS)
Johnson, Curtis W.; Anderson, William; Ross, Robert; Lyles, G. (Technical Monitor)
2000-01-01
Orbital Sciences Corporation has been awarded a contract by NASA's Marshall Space Flight Center, in cooperation with the U.S. Air Force Research Laboratory's Military Space Plane Technology Program Office, for the Upper Stage Flight Experiment (USFE) program. Orbital is designing, developing, and will flight test a new low-cost, 10,000 lbf hydrogen peroxide/ JP-8 pressure fed liquid rocket. During combustion chamber tests at NASA Stennis Space Center (SSC) of the USFE engine, the catalyst bed showed a low frequency instability occurring as the H202 flow reached about 1/3 its design rate. This paper reviews the USFE catalyst bed and combustion chamber and its operation, then discusses the dynamics of the instability. Next the paper describes the dynamic computer model used to recreate the instability. The model was correlated to the SSC test data, and used to investigate possible solutions to the problem. The combustion chamber configuration which solved the instability is shown, and the subsequent stable operation presented.
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77 FR 13257 - Submission for OMB Review; Comment Request
Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-06
..., Office of Information and Regulatory Affairs, Office of Management and Budget (OMB), Washington, DC, OIRA... Resource Management Plan (USFS 1990) and Upper McKenzie River Management Plan (``UMRMP,'' USFS 1992), (2...) inform management practices to protect and enhance the outstandingly remarkable values identified for the...
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Seal, S N; Davis, D L; Burch, J B
1991-05-01
The endogenous chicken vitellogenin II (VTGII) gene is transcribed exclusively in hepatocytes in response to estrogen. We previously identified two estrogen response elements (EREs) upstream of this gene. We now present an analysis of the VTGII promoter activated by these EREs in response to estrogen. Chimeric VTGII-CAT genes were cotransfected into LMH chicken hepatoma cells along with an estrogen receptor expression vector, and transient CAT expression was assayed after culturing the cells in the absence or presence of estrogen. An analysis of constructs bearing deletions downstream of the more proximal ERE indicated that promoter elements relevant to transcription in LMH cells extend to between -113 and -96. The relative importance of sequences within the VTGII promoter was examined by using 10 contiguous linker scanner mutations spanning the region from -117 to -24. Although most of these mutations compromised VTGII promoter function, one dramatically increased expression in LMH cells and also rendered the VTGII promoter capable of being activated by cis-linked EREs in fibroblasts cotransfected with an estrogen receptor expression vector. Gel retardation and DNase I footprinting assays revealed four factor-binding sites within this promoter. We demonstrate that three of these sites bind C/EBP, SP1, and USF (or related factors), respectively; the fourth site binds a factor that we denote TF-V beta. The biological relevance of these findings is suggested by the fact that three of these binding sites map to sites previously shown to be occupied in vivo in response to estrogen.
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78 FR 76789 - Additional Connect America Fund Phase II Issues
Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-19
... inspection and copying during normal business hours in the FCC Reference Information Center, Portals II, 445... Phase I to Phase II. 2. Timing of Phase II Support Disbursements. In the USF/ICC Transformation Order... language in paragraph 180 of the USF/ICC Transformation Order. We now seek to more fully develop the record...
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Acquaah-Mensah, George K.; Taylor, Ronald C.
Microarray data have been a valuable resource for identifying transcriptional regulatory relationships among genes. As an example, brain region-specific transcriptional regulatory events have the potential of providing etiological insights into Alzheimer Disease (AD). However, there is often a paucity of suitable brain-region specific expression data obtained via microarrays or other high throughput means. The Allen Brain Atlas in situ hybridization (ISH) data sets (Jones et al., 2009) represent a potentially valuable alternative source of high-throughput brain region-specific gene expression data for such purposes. In this study, Allen BrainAtlasmouse ISH data in the hippocampal fields were extracted, focusing on 508 genesmore » relevant to neurodegeneration. Transcriptional regulatory networkswere learned using three high-performing network inference algorithms. Only 17% of regulatory edges from a network reverse-engineered based on brain region-specific ISH data were also found in a network constructed upon gene expression correlations inmousewhole brain microarrays, thus showing the specificity of gene expression within brain sub-regions. Furthermore, the ISH data-based networks were used to identify instructive transcriptional regulatory relationships. Ncor2, Sp3 and Usf2 form a unique three-party regulatory motif, potentially affecting memory formation pathways. Nfe2l1, Egr1 and Usf2 emerge among regulators of genes involved in AD (e.g. Dhcr24, Aplp2, Tia1, Pdrx1, Vdac1, andSyn2). Further, Nfe2l1, Egr1 and Usf2 are sensitive to dietary factors and could be among links between dietary influences and genes in the AD etiology. Thus, this approach of harnessing brain region-specific ISH data represents a rare opportunity for gleaning unique etiological insights for diseases such as AD.« less
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Techniques to Improve Ultrasound-Switchable Fluorescence Imaging
NASA Astrophysics Data System (ADS)
Kandukuri, Jayanth
Novel approaches to the improvement of ultrasound-switchable fluorescence (USF) imaging--a relatively new imaging modality that combines ultrasound and optical imaging techniques--have been proposed for early cancer detection. In USF, a high-intensity focused ultrasound (HIFU) beam is used to induce temperature rise within its acoustic focal region due to which a thermo-sensitive USF contrast agent undergoes a switch in its state by increasing the output of fluorescence photons. By using an increase in fluorescence, one can isolate and quantify the fluorescence properties within the ultrasonic focal area. Therefore, USF is able to provide fluorescence contrast while maintaining ultrasound resolution in tissue. The major challenge of the conventional USF technique is its low axial resolution and its sensitivity (i.e. its signal-to-noise ratio (SNR)). This work focuses on investigating and developing a novel USF system design that can improve the resolution and SNR of USF imaging for biological applications. This work can be divided into two major parts: characterizing the performance of a high-intensity focused ultrasound transducer; and improving the axial resolution and sensitivity of the USF technique. Preliminary investigation was conducted by using an IR camera setup to detect temperature variation and thereby study the performance of the high-intensity focused ultrasound transducer to quantify different parameters of ultrasound-induced temperature focal size (UTFS). Investigations are conducted for the purpose of high-resolution imaging with an emphasis on HIFU-induced thermal focus size, short duration of HIFU-induced temperature increase (to avoid thermal diffusion or conduction), and control of HIFU-induced temperature increase within a few degrees Celsius. Next, the focus was shifted to improving the sensitivity of the ultrasound-switchable fluorescence-imaging technique. In this study, the USF signal is encoded with the modulation frequency of the ultrasound by modulating the induced temperature. Later, two approaches were adopted to modify the USF design to improve the resolution of the conventional USF imaging technique. The first approach aims to improve the axial resolution of conventional USF technique, which involves changing the USF system to adopt a dual-HIFU transducer arrangement (in which the transducers are 90 degree with respect to each other) for use as the heating source. The overlapped region of the two crossed foci (OR-TCF) of the dual-HIFU transducer module is expected to have small thermal size along both lateral and axial directions; thus, it could improve the axial resolution of the USF imaging technique. The second approach aims to demonstrate the improvement of resolution via a single-element HIFU transducer with a high frequency (15 MHz). The high frequency of the ultrasound transducer would have smaller acoustic lateral and axial size and should therefore have smaller thermal size. Thus, both approaches should be able to reduce the focal region of heating and thereby improve the resolution of the USF imaging. Results show that the driving power and exposure time of the HIFU transducer significantly influence the ultrasound-induced temperature focal size (UTFS). Interestingly, a nonlinear acoustic effect was observed at certain variations of the ultrasound exposure power while satisfying the thermal confinement within UTFS. This has been shown to reduce UTFS beyond the acoustic diffraction limit, while the ultrasound-induced thermal energy, which is confined within the focal volume, can induce a desired peak-temperature increase of a few degrees. On other hand, after encoding the HIFU exposure and therefore the detected USF signal with a modulation frequency, the SNR (sensitivity) and full width at half maximum (FWHM) along the lateral direction of the USF image was calculated to be 114 and 0.95 mm for a micro-tube with an inner diameter of 0.31 mm (ID), respectively. In comparison, they are 95 and 1.1 mm when using a non-modulated conventional USF imaging technique. In the case of improving the axial resolution of USF imaging for a similar target size, the dual-HIFU USF design was able to achieve 1.07 and 1.5 mm along lateral (x ) and axial (z) directions, respectively. Adopting the second approach of using single 15 MHz HIFU transducer for USF imaging, the axial resolution was calculated to be 0.67+/-0.02 mm and 1.71+/-0.24 mm along lateral (x) and axial (z) directions, respectively. Thus, high-resolution ultrasound-switchable fluorescence with good sensitivity can be designed for biomedical applications.
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New generation ICG-based contrast agents for ultrasound-switchable fluorescence imaging
Yu, Shuai; Cheng, Bingbing; Yao, Tingfeng; Xu, Cancan; Nguyen, Kytai T.; Hong, Yi; Yuan, Baohong
2016-01-01
Recently, we developed a new technology, ultrasound-switchable fluorescence (USF), for high-resolution imaging in centimeter-deep tissues via fluorescence contrast. The success of USF imaging highly relies on excellent contrast agents. ICG-encapsulated poly(N-isopropylacrylamide) nanoparticles (ICG-NPs) are one of the families of the most successful near-infrared (NIR) USF contrast agents. However, the first-generation ICG-NPs have a short shelf life (<1 month). This work significantly increases the shelf life of the new-generation ICG-NPs (>6 months). In addition, we have conjugated hydroxyl or carboxyl function groups on the ICG-NPs for future molecular targeting. Finally, we have demonstrated the effect of temperature-switching threshold (Tth) and the background temperature (TBG) on the quality of USF images. We estimated that the Tth of the ICG-NPs should be controlled at ~38–40 °C (slightly above the body temperature of 37 °C) for future in vivo USF imaging. Addressing these challenges further reduces the application barriers of USF imaging. PMID:27775014
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University of South Florida System Annual Accountability Report, 2013-14
ERIC Educational Resources Information Center
Board of Governors, State University System of Florida, 2014
2014-01-01
This statistical report provides data tables on the University of South Florida System's (USF's) financial resources, personnel, enrollment, undergraduate education, graduate education, and research & economic development. Highlights of USF's achievements in the 2013-2014 academic year include: (1) In Tampa, USF celebrated its strongest…
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76 FR 80941 - Request for Connect America Fund Cost Models
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-27
... interested parties to submit forward-looking cost models, consistent with the USF/ICC Transformation Order... forward-looking cost model consistent with the USF/ICC Transformation Order no later than December 30.... SUPPLEMENTARY INFORMATION: 1. On November 18, 2011, the Commission released the USF/ICC Transformation Order, 76...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-06-02
.... 8-9). The extent of C. vinaceum habitat on private property inholdings (privately owned land within... identified, of which 58 were on USFS land (Service 1993, p. 2). In 1995, there were 77 sites known to occur... species in 1987, surveys of USFS land estimated Cirsium vinaceum to be a species with 10,000 to 15,000...
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University of South Florida Libraries Search Committee Procedure Handbook.
ERIC Educational Resources Information Center
Heilos, Lawrence J.; And Others
This handbook of procedures developed by the Committee on Professional Concerns (COPC) of the University of South Florida (USF) describes the process to be used in recruiting and hiring qualified candidates for positions on the USF library faculty. The publication is divided into six sections: (1) information on the USF equal employment…
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Kandukuri, Jayanth; Yu, Shuai; Cheng, Bingbing; Bandi, Venugopal; D’Souza, Francis; Nguyen, Kytai T.; Hong, Yi; Yuan, Baohong
2017-01-01
Simultaneous imaging of multiple targets (SIMT) in opaque biological tissues is an important goal for molecular imaging in the future. Multi-color fluorescence imaging in deep tissues is a promising technology to reach this goal. In this work, we developed a dual-modality imaging system by combining our recently developed ultrasound-switchable fluorescence (USF) imaging technology with the conventional ultrasound (US) B-mode imaging. This dual-modality system can simultaneously image tissue acoustic structure information and multi-color fluorophores in centimeter-deep tissue with comparable spatial resolutions. To conduct USF imaging on the same plane (i.e., x-z plane) as US imaging, we adopted two 90°-crossed ultrasound transducers with an overlapped focal region, while the US transducer (the third one) was positioned at the center of these two USF transducers. Thus, the axial resolution of USF is close to the lateral resolution, which allows a point-by-point USF scanning on the same plane as the US imaging. Both multi-color USF and ultrasound imaging of a tissue phantom were demonstrated. PMID:28165390
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Chromatin insulator elements: establishing barriers to set heterochromatin boundaries.
Barkess, Gráinne; West, Adam G
2012-02-01
Epigenomic profiling has revealed that substantial portions of genomes in higher eukaryotes are organized into extensive domains of transcriptionally repressive chromatin. The boundaries of repressive chromatin domains can be fixed by DNA elements known as barrier insulators, to both shield neighboring gene expression and to maintain the integrity of chromosomal silencing. Here, we examine the current progress in identifying vertebrate barrier elements and their binding factors. We overview the design of the reporter assays used to define enhancer-blocking and barrier insulators. We look at the mechanisms vertebrate barrier proteins, such as USF1 and VEZF1, employ to counteract Polycomb- and heterochromatin-associated repression. We also undertake a critical analysis of whether CTCF could also act as a barrier protein. There is good evidence that barrier elements in vertebrates can form repressive chromatin domain boundaries. Future studies will determine whether barriers are frequently used to define repressive domain boundaries in vertebrates.
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The USF Libraries Virtual Library Project: A Blueprint for Development.
ERIC Educational Resources Information Center
Metz-Wiseman, Monica; Silver, Susan; Hanson, Ardis; Johnston, Judy; Grohs, Kim; Neville, Tina; Sanchez, Ed; Gray, Carolyn
This report of the Virtual Library Planning Committee (VLPC) is intending to serve as a blueprint for the University of South Florida (USF) Libraries as it shifts from print to digital formats in its evolution into a "Virtual Library". A comprehensive planning process is essential for the USF Libraries to make optimum use of technology,…
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Crime in woods: role of law enforcement officers in national forests
Joanne F. Tynon; Deborah J. Chavez; Joshua W. R. Baur
2010-01-01
This first nationwide study of US Forest Service (USFS) law enforcement officers (LEOs) examined respondentsâ roles in the USFS, what they perceived as their highest work priority, and what their relationship with the rest of the USFS should be. Results show that LEOs believe they have a high priority for protecting forest users and they believe that National Forest...
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NASA Technical Reports Server (NTRS)
Spurce, Joseph P.; Hargrove, William; Ryan, Robert E.; Smooth, James C.; Prados, Don; McKellip, Rodney; Sader, Steven A.; Gasser, Jerry; May, George
2008-01-01
This viewgraph presentation reviews a project, the goal of which is to study the potential of MODIS data for monitoring historic gypsy moth defoliation. A NASA/USDA Forest Service (USFS) partnership was formed to perform the study. NASA is helping USFS to implement satellite data products into its emerging Forest Threat Early Warning System. The latter system is being developed by the USFS Eastern and Western Forest Threat Assessment Centers. The USFS Forest Threat Centers want to use MODIS time series data for regional monitoring of forest damage (e.g., defoliation) preferably in near real time. The study's methodology is described, and the results of the study are shown.
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NASA Astrophysics Data System (ADS)
Mayo, M.; Ithier-Guzman, W.; Pyrtle, A. J.; Betzer, P.; Batson, B.; Bhansali, S.; Greene, B.; Turner, R.
2007-05-01
In 2004, the University of South Florida (USF) was granted by the National Science Foundation a Louis Stokes Alliance for Minority Participation (LSAMP) Bridge to the Doctorate (BD) site award (HRD# 0217675). As part of the Florida-Georgia Louis Stokes Alliance for Minority Participation (FGLSAMP), USF is one of thirteen institutions in an alliance that is dedicated to significantly increasing the number of underrepresented minority students who obtain undergraduate and graduate STEM degrees. The BD program at USF incorporates the goals of FGLSAMP and facilitates the recruitment of underrepresented minorities pursuing careers in the STEM fields at the graduate level. The thematic focus of the FGLSAMP USF BD program is focused on the development and application of biogeochemical sensors for marine, aquatic, environmental, remote sensing and biomedical applications. After recruitment, BD graduate fellowship recipients are provided with NSF-funded financial support for two years, and opportunities to participate in professional development workshops, seminars and short courses, as well as additional financial support to pursue and complete their doctoral studies (beyond the initial two years of NSF BD funding), in a variety of forms, including, but not limited to, Alfred P. Sloan Minority Scholarships, Florida Education Fund's McKnight Doctoral Fellowships, USF College of Graduate Studies Fellowships, USF CMS endowed fellowships, USF CMS research assistantships, and USF CMS teaching assistantships. Collectively, 3 LSAMP BD grants have been awarded at USF to support 56 underrepresented minority fellowship recipients, of which 14 are currently graduate students at the USF College of Marine Science (CMS). Since the arrival of the BD Fellowship program, the graduate community has diversified, showing an increase of over 40% in underrepresented minorities at CMS. The BD program has enhanced the research and learning environment for all CMS students, as well as fostered a nurturing community of underrepresented minority CMS graduate students committed to obtaining their doctoral degrees. As of spring 2007, a total of 4 BD fellowship recipients have obtained marine science master's degrees and are currently pursuing their doctoral degrees in the CMS. In addition, in less than two years, a BD endowment fund of more than $900,000 was established. This fund will provide financial support for at least two minority CMS graduate students in perpetuity! Lastly, in response to an identified need for increased ocean literacy among underrepresented groups, several BD fellowship recipients have engaged in activities designed to "give back" via informal and formal education and outreach opportunities within their native communities.
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Fetal iron deficiency induces chromatin remodeling at the Bdnf locus in adult rat hippocampus.
Tran, Phu V; Kennedy, Bruce C; Lien, Yu-Chin; Simmons, Rebecca A; Georgieff, Michael K
2015-02-15
Fetal and subsequent early postnatal iron deficiency causes persistent impairments in cognitive and affective behaviors despite prompt postnatal iron repletion. The long-term cognitive impacts are accompanied by persistent downregulation of brain-derived neurotrophic factor (BDNF), a factor critical for hippocampal plasticity across the life span. This study determined whether early-life iron deficiency epigenetically modifies the Bdnf locus and whether dietary choline supplementation during late gestation reverses these modifications. DNA methylation and histone modifications were assessed at the Bdnf-IV promoter in the hippocampus of rats [at postnatal day (PND) 65] that were iron-deficient (ID) during the fetal-neonatal period. Iron deficiency was induced in rat pups by providing pregnant and nursing dams an ID diet (4 mg/kg Fe) from gestational day (G) 2 through PND7, after which iron deficiency was treated with an iron-sufficient (IS) diet (200 mg/kg Fe). This paradigm resulted in about 60% hippocampal iron loss on PND15 with complete recovery by PND65. For choline supplementation, pregnant rat dams were given dietary choline (5 g/kg) from G11 through G18. DNA methylation was determined by quantitative sequencing of bisulfite-treated DNA, revealing a small alteration at the Bdnf-IV promoter. Chromatin immunoprecipitation analysis showed increased HDAC1 binding accompanied by reduced binding of RNA polymerase II and USF1 at the Bdnf-IV promoter in formerly ID rats. These changes were correlated with altered histone methylations. Prenatal choline supplementation reverses these epigenetic modifications. Collectively, the findings identify epigenetic modifications as a potential mechanism to explicate the long-term repression of Bdnf following fetal and early postnatal iron deficiency. Copyright © 2015 the American Physiological Society.
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Fetal iron deficiency induces chromatin remodeling at the Bdnf locus in adult rat hippocampus
Kennedy, Bruce C.; Lien, Yu-Chin; Simmons, Rebecca A.; Georgieff, Michael K.
2014-01-01
Fetal and subsequent early postnatal iron deficiency causes persistent impairments in cognitive and affective behaviors despite prompt postnatal iron repletion. The long-term cognitive impacts are accompanied by persistent downregulation of brain-derived neurotrophic factor (BDNF), a factor critical for hippocampal plasticity across the life span. This study determined whether early-life iron deficiency epigenetically modifies the Bdnf locus and whether dietary choline supplementation during late gestation reverses these modifications. DNA methylation and histone modifications were assessed at the Bdnf-IV promoter in the hippocampus of rats [at postnatal day (PND) 65] that were iron-deficient (ID) during the fetal-neonatal period. Iron deficiency was induced in rat pups by providing pregnant and nursing dams an ID diet (4 mg/kg Fe) from gestational day (G) 2 through PND7, after which iron deficiency was treated with an iron-sufficient (IS) diet (200 mg/kg Fe). This paradigm resulted in about 60% hippocampal iron loss on PND15 with complete recovery by PND65. For choline supplementation, pregnant rat dams were given dietary choline (5 g/kg) from G11 through G18. DNA methylation was determined by quantitative sequencing of bisulfite-treated DNA, revealing a small alteration at the Bdnf-IV promoter. Chromatin immunoprecipitation analysis showed increased HDAC1 binding accompanied by reduced binding of RNA polymerase II and USF1 at the Bdnf-IV promoter in formerly ID rats. These changes were correlated with altered histone methylations. Prenatal choline supplementation reverses these epigenetic modifications. Collectively, the findings identify epigenetic modifications as a potential mechanism to explicate the long-term repression of Bdnf following fetal and early postnatal iron deficiency. PMID:25519736
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Worldwide trends in Universal Service Funds and telemedicine.
Nakajima, Isao
2010-12-01
A survey of recent worldwide trends in Universal Service Funds (USFs) and the assistance provided for their application indicates that industrialized countries and developing nations alike have offered or plan to offer tax-relief measures or reimbursement for communications costs incurred by telemedicine programs, thus finding a way to actively apply USFs in rural areas. There are three main systems used to calculate the amount of reimbursement from a USF. While many countries adopt a service-area net-loss estimation method, Japan uses a benchmark method and provides financial assistance only to unprofitable areas. The USA has proactively introduced telemedicine to rural areas and isolated islands in order to minimize rapidly rising healthcare costs and to improve the efficiency of healthcare services. In the USA, the USF is used to pay back communications costs incurred through telemedicine programs. For instance, the budget allocated from the USF for reimbursements for telemedicine in Alaska reached USD 30 Mil. in 2007. Developing countries in Africa and Asia are operating various forms of telemedicine on a trial basis, but a tax-relief measure or payback of communications costs, which are a large portion of the running costs, will need to be implemented to ensure sustainable and autonomous operation of telemedicine. In Japan, up until January 2007, the USF system assumed the use of an NTS (non-traffic sensitive cost) system to obtain funds from connection fees, and this system would receive funds from each telecommunications carrier (payer: the telecommunications carriers). The beneficiaries would be limited to two companies, namely NTT East and NTT West. However, the Japanese USF system was revised in February 2007, and a fee is now collected from each telephone number (payer: the user). The collected funds are used to cover losses in unprofitable areas (not limited to remote areas) among 7,000 business areas in Japan. In view of worldwide trends, the author believes that Japan should also start using the USF system to reimburse communications costs (including costs of telemedicine) in order to achieve sustainable and autonomous operation of public communication systems in rural areas.
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PHOTOCOPY OF STANDARD USDA/USFS PLAN FOR 13' X 13' STEEL ...
PHOTOCOPY OF STANDARD USDA/USFS PLAN FOR 13' X 13' STEEL LOOKOUT HOUSE (CAB); ELEVATIONS, SECTIONS, MISC. DETAILS; DATED 1961 - North Mountain Lookout, Stanislaus National Forest, Groveland, Tuolumne County, CA
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Deep patch technique for landslide repair. Final report
DOE Office of Scientific and Technical Information (OSTI.GOV)
Helwany, B.M.
1994-10-01
The report describes the laboratory testing of the `USFS deep patch` technique and a CTI modification of this technique for repairing landslides with geosynthetic reinforcement. The technique involves replacing sections of roadway lost due to landslides on top of a geosynthetically-reinforced embankment. The CTI modification involves replacing the reinforced slope with a geosynthetically-reinforced retaining wall with a truncated base. Both techniques rely on the cantilevering ability of the reinforced mass to limit the load on the foundation with a high slide potential. The tests with road base showed that (1) both the USFS and CTI repair reduced effectively the adversemore » effects of local landsliding on the highway pavement by preventing crack propagation; (2) the USFS repair increased the stability of the repaired slope, which was in progressive failure, by reducing the stresses exerted on it; and (3) the CTI repair produced substantially greater stresses on its foundation due to the truncated base of the reinforced mass.« less
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What is the best cryopreservation protocol for human testicular tissue banking?
Baert, Y; Van Saen, D; Haentjens, P; In't Veld, P; Tournaye, H; Goossens, E
2013-07-01
Is there a better alternative to the conventional cryopreservation protocols for human testicular tissue banking? Uncontrolled slow freezing (USF) using 1.5 M dimethylsulphoxide (DMSO) and 0.15 M sucrose as cryoprotectants appears to be a user-friendly and efficient method for the cryopreservation of human testicular tissue. Currently, time-consuming controlled slow freezing (CSF) protocols that need expensive equipment are commonly used for human testicular tissue banking. USF and vitrification are cryopreservation techniques that were successfully applied in several animal models but need further exploration with human tissue. Fragments (n = 160) of testicular tissue from 14 patients undergoing vasectomy reversal were assigned to a fresh control group or one of the following cryopreservation procedures: CSF using DMSO at a concentration of 0.7 or 1.5 M in the presence (+S) or absence of sucrose (-S), USF using either 0.7 or 1.5 M DMSO combined with sucrose, solid-surface vitrification (SSV) or direct cover vitrification (DCV). Light microscopic evaluations were performed to study apoptosis, germ cell proliferation ability, spermatogonial survival, coherence of the seminiferous epithelium and integrity of the interstitial compartment after cryopreservation. Ultrastructural alterations were studied by scoring cryodamage to four relevant testicular cell types. The USF 1.5 M DMSO + S protocol proved not solely to prevent cell death and to preserve seminiferous epithelial coherence, interstitial compartment integrity, SG and their potential to divide but also protected the testicular cell ultrastructure. A significant reduction in the number of SG per tubule from 21.4 ± 5.6 in control tissue to 4.9 ± 2.1, 8.2 ± 5.4, 11.6 ± 5.1, 8.8 ± 3.9, 12.6 ± 4.4 and 11.7 ± 5.7 was observed after cryopreservation combined with at least one other form of cryoinjury when using CSF 0.7 M DMSO -S, CSF 0.7 M DMSO + S, CSF 1.5 M DMSO + S, USF 0.7 M DMSO + S, SSV and direct cover vitrification (DCV), respectively (P < 0.001). Supplementary research is required to investigate the effect on tissue functionality and to confirm this study's findings using prepubertal tissue. An optimal cryopreservation protocol enhances the chances for successful fertility restoration. USF, being an easy and cost-effective alternative to CSF, would be preferable for laboratories in developing countries or whenever tissue is to be procured from a diseased child at a site distant from the banking facility.
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77 FR 11109 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-24
...-Native Plant Control Project, Proposes a Forest-Wide Integrated Management Strategy to Control the Spread..., Final EIS, USFS, SD, Steamboat Project, Proposes to Implement Multiple Resource Management Actions... EIS, USFS, WA, South George Vegetation and Fuels Management Project, To Improve Forest Health and...
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78 FR 32991 - Connect America Fund
Federal Register 2010, 2011, 2012, 2013, 2014
2013-06-03
..., 2013. The full text of this document is available for public inspection during regular business hours.... Introduction 1. In the USF/ICC Transformation Order, 76 FR 73830, November 29, 2011, the Commission... the USF/ICC Transformation Order, an unsubsidized competitor in areas where the price cap carrier will...
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S. Karen Dante-Wood
2018-01-01
The Northern Rockies Adaptation Partnership (NRAP) is a science-management partnership among the Forest Service, U.S. Department of Agriculture (USFS) regional offices and national forests (mostly in the Northern Region, and small portions of the Intermountain and Rocky Mountain Regions); USFS Pacific Northwest and Rocky Mountain Research Stations; Glacier, Yellowstone...
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Kandt, Alicen J.; Kiatreungwattana, Kosol
This report summarizes the results from an energy efficiency, water efficiency, and renewable energy site assessment of the Dolores Ranger District in the San Juan National Forest in Colorado. A team led by the U.S. Department of Energy's National Renewable Energy Laboratory (NREL) conducted the assessment with United States Forest Service (USFS) personnel on August 16-17, 2016, as part of ongoing efforts by USFS to reduce energy and water use and implement renewable energy technologies. The assessment is approximately an American Society of Heating, Refrigerating, and Air-Conditioning Engineers Level 2 audit and meets Energy Independence and Security Act requirements.
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Fire management assessment of Eastern Province, Zambia
L. T. Hollingsworth; D. Johnson; G. Sikaundi; S. Siame
2015-01-01
The mission that produced this assessment was prompted by requests from Forestry Department personnel in Zambia to the United States Agency for International Development (USAID) for formal fire management training. USAID contacted the United States Forest Service's (USFS) International Programs (IP) with the training request. Together, USFS, USAID, and Zambian...
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Joanne J. Ho
2018-01-01
The Intermountain Adaptation Partnership (IAP) is a science-management partnership with a wide variety of participants across the U.S. Department of Agriculture Forest Service (USFS) Intermountain Region, which spans Nevada, Utah, southern Idaho, eastern California, and western Wyoming. This USFS region is the largest in the Nation, representing nearly 17 percent of...
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Florida Public Health Training Center: Evidence-Based Online Mentor Program
ERIC Educational Resources Information Center
Frahm, Kathryn A.; Alsac-Seitz, Biray; Mescia, Nadine; Brown, Lisa M.; Hyer, Kathy; Liburd, Desiree; Rogoff, David P.; Troutman, Adewale
2013-01-01
This article describes an Online Mentor Program (OMP) designed to support and facilitate mentorships among and between Florida Department of Health (FDOH) employees and USF College of Public Health students using a Web-based portal. The Florida Public Health Training Center (FPHTC) at the University of South Florida (USF) College of Public Health…
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William H. Butler; Ashley Monroe; Sarah McCaffrey
2015-01-01
The Collaborative Forest Landscape Restoration Program (CFLRP), established in 2009, encourages collaborative landscape scale ecosystem restoration efforts on United States Forest Service (USFS) lands. Although the USFS employees have experience engaging in collaborative planning, CFLRP requires collaboration in implementation, a domain where little prior experience...
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A Study of Instructional Faculty Salaries at USF, SUS and National Peers
ERIC Educational Resources Information Center
Micceri, Theodore
2010-01-01
This study investigates 10-year trends in instructional faculty salaries by sex and rank for USF, five SUS Peers (UF, FSU, FIU, UCF, FAU) and eight National Peers (North Carolina State, Alabama-Birmingham, Illinois-Chicago, California-Irvine, SUNY-Stony Brook, SUNY-Buffalo, Cincinnati, Rutgers). Methods: Historical instructional faculty salary…
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View west from USFS Road 369 toward Fowler Lode Adit ...
View west from USFS Road 369 toward Fowler Lode Adit and O'Brien Ditch (left) and open-pit excavation (top-center, beyond trees); ridge saddle is in center - Steamboat Mine, Southeast slope of Steamboat Mountain, west of the junction of Forest Service Roads 1000300 and 1000365, Jacksonville, Jackson County, OR
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USDA Forest Service goals and programs for monitoring neotropical migratory birds
Patricia Manley
1993-01-01
The USDA Forest Service (USFS) developed goals, objectives, and guidelines for monitoring neotropical migratory birds (NTMB) on National Forest System lands in response to the Neotropical Migratory Bird Conservation Program Partners in Flight. A USFS task group developed a hierarchical monitoring framework designed to define priorities for type of monitoring data....
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ERIC Educational Resources Information Center
Lim, Victoria
2008-01-01
This article describes how the benefactors of University of South Florida (USF) Latino Scholarship Program (LSP) support students in more ways than one. LSP, now in its 17th year, was started by the Latin Community Advisory Committee to the USF president to attract Hispanic students to the school. The committee didn't want to just provide…
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Defining Old Growth: Implications For Management
David L. White; F. Thomas Lloyd
1994-01-01
USDA Forest Service (USFS), with the help of scientists from The Nature Conservancy (TNC), Forest Service Research and ther organizations, is developing old-growth definitions for 35 forest types within the Eastern United States (U.S.). Old-growth forests were officially recognized as a resource by the USFS in 1988 and shortly thereafter, the Eastern Old-Growth...
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75 FR 68355 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-05
... District Olympic National Forest, Olympic National Park, Jefferson County, WA, Wait Period Ends: 12/06/2010...: Lynne Urquhart 334-274-6371. EIS No. 20100431, Final EIS, USFS, WA, Dosewallips Road Washout Project, To...: 12/20/2010, Contact: Amy Henry 865-632-4045. EIS No. 20100433, Final EIS, USFS, CA, Plumas National...
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National FIA plot intensification procedure report
Jock A. Blackard; Paul L. Patterson
2014-01-01
The Forest Inventory and Analysis (FIA) program of the U.S. Forest Service (USFS) measures a spatially distributed base grid of forest inventory plots across the United States. The sampling intensity of plots may be increased in some regions when warranted by specific inventory objectives. Several intensification methods have been developed within FIA and USFS National...
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The Applegate Adaptive Management Area ecosystem health assessment
Thomas Atzet
1995-01-01
As requested by the Applegate Partnership, the Medford District Bureau of Land Management, the Rogue River and Siskiyou National Forests, a team of six specialists (Dr. Tom Atzet, USFS ecologist; Dr. Mike Amaranthus, PNW soil scientist, Dr. Don Goheen, USFS pathologist and entomologist, Tom Sensenig, BLM silviculturist, Dr. Dave Perry, Oregon State University...
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Chapter 17: Forecasting wildfire suppression expenditures for the United States Forest Service
Karen L. Abt; Jeffrey P. Prestemon; Krista Gebert
2008-01-01
The wildland fire management organization of the United States Forest Service (USFS) operates under policy and budget legacies that began nearly 100 years ago and a forest fuel situation that is all too current. The confluence of these three factors contributes to increased burning and fire fighting costs for the agency, and increased concern from both the U.S....
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Forecasting wildfire suppression expenditures for the United States Forest Service
Karen L. Abt; Jeffrey P. Prestemon; Krista Gebert
2008-01-01
The wildland fire management organization of the United States Forest Service (USFS) operates under policy and budget legacies that began nearly -100 years ago and a forest fuel situation that is all too current. The confluence of these three factors contributes to increased burning and firefighting costs for the agency, and increased concern from both the U.S....
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Learning from wilderness: The social dimension of fire management
Anne E. Black
2009-01-01
In 2008, the U.S. Forest Service (USFS) began piloting a "new" concept in fire management: managing "fire as fire" on the landscape; no more black-and-white distinctions between "good" fire and "bad" fire. Instead, under the new direction, the USFS manages the fire based on what the land, the long-term objectives, the land...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-06-06
... document is available for inspection and copying during normal business hours in the FCC Reference... carriers supports such networks, and indeed, under the USF/ICC Transformation Order, 76 FR 73830, November... originally sought comment on this proposal in the USF/ICC Transformation Order FNPRM, 76 FR 73830, November...
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ERIC Educational Resources Information Center
Molinari, Victor; Chiriboga, David A.; Schonfeld, Lawrence; Haley, William E.; Schinka, John A.; Hyer, Kathy; Dupree, Larry W.
2005-01-01
There is a growing need for geropsychologists who are specialists in practice, research, education, and advocacy for older adults. The combined USF/Tampa VA geropsychology fellowship program focuses on the training of three post-doctoral Fellows each year in public sector service delivery across diverse long term care (LTC) and primary care…
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Urban forest health monitoring: large-scale assessments in the United States
Anne Buckelew Cumming; Daniel B. Twardus; David J. Nowak
2008-01-01
The U.S. Department of Agriculture, Forest Service (USFS), together with state partners, developed methods to monitor urban forest structure, function, and health at a large statewide scale. Pilot studies have been established in five states using protocols based on USFS Forest Inventory and Analysis and Forest Health Monitoring program data collection standards....
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Evaluation of open source data mining software packages
Bonnie Ruefenacht; Greg Liknes; Andrew J. Lister; Haans Fisk; Dan Wendt
2009-01-01
Since 2001, the USDA Forest Service (USFS) has used classification and regression-tree technology to map USFS Forest Inventory and Analysis (FIA) biomass, forest type, forest type groups, and National Forest vegetation. This prior work used Cubist/See5 software for the analyses. The objective of this project, sponsored by the Remote Sensing Steering Committee (RSSC),...
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Land cover change map comparisons using open source web mapping technologies
Erik Lindblom; Ian Housman; Tony Guay; Mark Finco; Kevin Megown
2015-01-01
The USDA Forest Service is evaluating the status of current landscape change maps and assessing gaps in their information content. These activities have been occurring under the auspices of the Landscape Change Monitoring System (LCMS) project, which is a joint effort between USFS Research, USFS Remote Sensing Applications Center (RSAC), USGS Earth Resources...
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What are the best seed sources for ecosystem restoration on BLM and USFS lands?
Larry Stritch; Peggy Olwell; Scott Lambert; Matthew E. Horning; Richard Cronn
2010-01-01
Native plant restoration policy calls for use of "genetically appropriate" native plant material on USDI Bureau of Land Management (BLM) and USDA Forest Service (USFS) lands. In this article, we summarize experimental evidence showing that local adaptation is widespread in all kingdoms of life, and how this "home-field advantage" has been exploited...
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"It was a young man's life": G. A. Pearson
Susan D. Olberding
2008-01-01
The nation's initial USFS research site commenced in a rustic cabin in the midst of northern Arizona's expansive ponderosa pine forest. Gustaf A. Pearson was the first in a distinguished line of USFS scientists to live and study there. A visitor to Fort Valley today often wishes he could have stood in Pearson's large boots (he was said to have enormous...
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Crystal L. Raymond; David L. Peterson; Regina M. Rochefort
2013-01-01
The U.S. Forest Service (USFS) and National Park Service (NPS) have highlighted climate change as an agency priority and issued direction to administrative units for responding to climate change. In response, the USFS and NPS initiated the North Cascadia Adaptation Partnership (NCAP) in 2010. The goals of the NCAP were to build an inclusive partnership, increase...
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Riparian area protection and outdoor recreation: lessons from the Northwest Forest Plan
Patrick Impero Wilson; Troy E. Hall; Linda E. Kruger
2012-01-01
The Northwest Forest Plan required the US Forest Service (USFS) to shift its management focus to ecological values rather than the utilitarian ones that had dominated forest policy in the region. This article examines the effects of this shift on the USFS's historic mission to provide recreational access to the region's forests. Focusing on six national...
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Mostazir, M; Jeffery, A; Voss, L; Wilkin, T
2014-06-01
Body mass index (BMI) is reportedly gender assortative (mother-daughter, father-son) in contemporary children. We investigated the corresponding transmission of waist circumference (WC) and its implications. We measured parental WC at baseline and WC, height, weight and para-umbilical skin-fold (USF) annually in their offspring from 5 to 15 years (n = 223 trios). Parents were deemed normal metabolic risk (NR) or high risk (HR) according to World Health Organization (WHO) cut-points for WC (mothers 80 cm, fathers 94 cm). The residual from WC adjusted for BMI (WC|BMI ) was used as a surrogate for excess intra-abdominal fat, and its association with insulin resistance (HOMA2-IR) was sought. WC and USF were both gender assortative, while WC|BMI was not. WC was greater by 1.62 cm (P < 0.05, confidence interval [CI]: 0.09-3.16) and USF by 0.37 cm (P < 0.01, CI: 0.19-0.56) among the daughters (but not the sons) of HR compared with those of NR mothers, and by 1.32 cm (P < 0.05, CI: 0.09-2.55) and 0.18 cm (P < 0.05, CI: 0.04-0.32), respectively in the corresponding father-son (but not father-daughter) pairings. No such differences could be demonstrated for WC|BMI . A standard deviation score 1(SDS) change in WC|BMI , independent of BMI, was associated with a 7.14% change in IR in girls (P < 0.01, CI: 1.76-12.80) and 8.02% in boys (P < 0.001, CI: 2.93-13.36), but there was no relationship between IR and USF. The relationship of offspring WC to metabolic health and to parental size is complex. Subcutaneous abdominal fat is gender assortative but harmless, while intra-abdominal fat (its surrogate in this analysis) is unrelated to parental waist circumference, but metabolically harmful. © 2013 The Authors. Pediatric Obesity © 2013 International Association for the Study of Obesity.
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76 FR 59125 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-09-23
.... Markham 805-585-2150. EIS No. 20110317, Draft EIS, USFS, MT, Lonesome Wood Vegetation Management 2 Project... the 2006 FEIS Analysis and to Correct the Deficiencies that the Meister Panel Identified, Land and...
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Conterminous U.S. and Alaska Forest Type Mapping Using Forest Inventory and Analysis Data
B. Ruefenacht; M.V. Finco; M.D. Nelson; R. Czaplewski; E.H. Helmer; J. A. Blackard; G.R. Holden; A.J. Lister; D. Salajanu; D. Weyermann; K. Winterberger
2008-01-01
Classification-trees were used to model forest type groups and forest types for the conterminous United States and Alaska. The predictor data were a geospatial data set with a spatial resolution of 250 m developed by the U.S. Department of Agriculture Forest Service (USFS). The response data were plot data from the USFS Forest Inventory and Analysis program. Overall...
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Management perceptions of off-highway vehicle use on national forest system lands in Appalachia
Katherine A. Thompson; Chad D. Pierskalla; Michael A. Schuett
2008-01-01
In 2005, the U.S. Forest Service (USFS) issued new standards for dealing with unmanaged recreation. All National Forest System units are to develop travel management plans by 2009. The purpose of this study was to determine differences in perceptions between USFS managers of national forests in Appalachia with low and high levels of offhighway vehicle (OHV) use...
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Forest Service Groundwater Plan Oversteps Bounds, Critics Say
NASA Astrophysics Data System (ADS)
Showstack, Randy
2014-09-01
The U.S. Forest Service (USFS) is at odds with some members of Congress and other critics about a proposed government directive on groundwater resource management. The USFS says the proposed directive is an innocuous internal measure to provide a consistent, systematic, and transparent agency-wide approach to groundwater management. However, some participants in a 10 September congressional hearing questioned the directive, saying that the agency is overstepping its bounds.
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"It was a young man's life": G.A. Pearson (P-53)
Susan D. Olberding
2008-01-01
The nation's initial USFS research site commenced in a rustic cabin in the midst of northern Arizonaâs expansive ponderosa pine forest. Gustaf A. Pearson was the first in a distinguished line of USFS scientists to live and study there. A visitor to Fort Valley today often wishes he could have stood in Pearson's large boots (he was said to have enormous feet)...
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R. Hutto; Skip Kowalski
2006-01-01
The Northern Region Landbird Monitoring Program (NRLMP) began in 1990 as a cooperative effort between the United States Forest Service (USFS) and the University of Montana. The combination of a research-oriented perspective from the University and a management-needs perspective from the National Forests within the Northern Region led to the realization that landbirds...
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Robbins, Lisa L.; Knorr, Paul O.; Daly, Kendra L.; Barrera, Kira E.
2014-01-01
During September and November 2011 the (USGS), in cooperation with (USF), conducted geochemical surveys on the west Florida Shelf to investigate the effects of climate change on ocean acidification within the northern Gulf of Mexico, specifically, the effect of ocean acidification on marine organisms and habitats. The first cruise was conducted from September 20 to 28 (11BHM03) and the second was from November 2 to 4 (11BHM04). To view each cruise's survey lines, please see the Trackline page. Each cruise took place aboard the Research Vessel (R/V) Weatherbird II, a ship of opportunity led by Dr. Kendra Daly (USF), which departed from and returned to Saint Petersburg, Florida. Data collection included sampling of the surface and water column with lab analysis of pH, dissolved inorganic carbon (DIC) or total carbon dioxide (TCO2), and total alkalinity (TA). lLb analysis was augmented with a continuous flow-through system (referred to as sonde data) with a conductivity-temperature-depth (CTD) sensor, which also recorded salinity and pH. Corroborating the USGS data are the vertical CTD profiles (referred to as station samples) collected by USF. The CTD casts measured continuous vertical profiles of oxygen, chlorophyll fluorescence and optical backscatter. Discrete samples for nutrients, chlorophyll, and particulate organic carbon/nitrogen were also collected during the CTD casts. Two autonomous flow-through (AFT) instruments recorded pH and CO2 every 3-5 minutes on each cruise (referred to as AFT data).
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Learning to Live with Off-Highway Vehicles: Lessons Learned from the Dixie National Forest
Aaron K. Divine; Pamela E. Foti
2004-01-01
Nationwide, there are an estimated 446,000 miles of road on United States Forest Service (USFS) landsâfour times that of any other public land management agency (USDA 2000; Havlick 2002). Most USFS roads were developed as part of a network to access timber on some 192 million acres of forested land during the past century (Forman et al. 2003). In recent years,...
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Southern California Disasters II
NASA Technical Reports Server (NTRS)
Nicholson, Heather; Todoroff, Amber L.; LeBoeuf, Madeline A.
2015-01-01
The USDA Forest Service (USFS) has multiple programs in place which primarily utilize Landsat imagery to produce burn severity indices for aiding wildfire damage assessment and mitigation. These indices provide widely-used wildfire damage assessment tools to decision makers. When the Hyperspectral Infrared Imager (HyspIRI) is launched in 2022, the sensor's hyperspectral resolution will support new methods for assessing natural disaster impacts on ecosystems, including wildfire damage to forests. This project used simulated HyspIRI data to study three southern California fires: Aspen, French, and King. Burn severity indices were calculated from the data and the results were quantitatively compared to the comparable USFS products currently in use. The final results from this project illustrate how HyspIRI data may be used in the future to enhance assessment of fire-damaged areas and provide additional monitoring tools for decision support to the USFS and other land management agencies.
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Robbins, Lisa L.; Knorr, Paul O.; Daly, Kendra L.; Taylor, Carl A.; Barrera, Kira E.
2014-01-01
During May and June 2011 the (USGS), in cooperation with (USF), conducted geochemical surveys on the west Florida Shelf to investigate the effects of climate change on ocean acidification within the northern Gulf of Mexico, specifically, the effect of ocean acidification on marine organisms and habitats. The first cruise was conducted from May 3 to 9 (11BHM01) and the second was from June 25 to 30 (11BHM02). To view each cruise's survey lines, please see the Trackline page. Each cruise took place aboard the Research Vessel (R/V) Weatherbird II, a ship of opportunity led by Dr. Kendra Daly (USF), which departed from and returned to Saint Petersburg, Florida. Data collection included sampling of the surface and water column with lab analysis of pH, dissolved inorganic carbon (DIC) or total carbon dioxide (TCO2), and total alkalinity (TA). lLb analysis was augmented with a continuous flow-through system (referred to as sonde data) with a conductivity-temperature-depth (CTD) sensor, which also recorded salinity and pH. Corroborating the USGS data are the vertical CTD profiles (referred to as station samples) collected by USF. The CTD casts measured continuous vertical profiles of oxygen, chlorophyll fluorescence and optical backscatter. Discrete samples for nutrients, chlorophyll, and particulate organic carbon/nitrogen were also collected during the CTD casts. Two autonomous flow-through (AFT) instruments recorded pH and CO2 every 3-5 minutes on each cruise (referred to as AFT data).
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2007-09-30
marine.usf.edu Aida Alvera -Azcárate Phone: (727) 553-3508 Fax: (727) 553 1189 email: aalvera@marine.usf.edu Alexander Barth, Phone: (727...WFS and the data assimilation are being performed by Alexander Barth. The Cariaco nesting is performed by Aida Alvera -Azcárate. WORK COMPLETED...363–380. Barth, A., Beckers, J.-M., Alvera -Azcárate, A. and Weisberg, R.H. (2007), Filtering inertia-gravity waves from the initial conditions of
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2009-06-01
USACE 2008c)) on June 3, 1992 that “effectively precludes the future use of expanded polystyrene unless it is encased in an approved protective coating...punctured. Closed cell (extruded) expanded polystyrene of good quality and manufac- tured for marine use will be required. Lesser quality foam bead flota...Forest Service (USFS) (USFS 2008) – “Open cell Expanded Polystyrene Foam (EPS) has an open structure that easily lets water into its interior. It
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Statistical studies of animal response data from USF toxicity screening test method
NASA Technical Reports Server (NTRS)
Hilado, C. J.; Machado, A. M.
1978-01-01
Statistical examination of animal response data obtained using Procedure B of the USF toxicity screening test method indicates that the data deviate only slightly from a normal or Gaussian distribution. This slight departure from normality is not expected to invalidate conclusions based on theoretical statistics. Comparison of times to staggering, convulsions, collapse, and death as endpoints shows that time to death appears to be the most reliable endpoint because it offers the lowest probability of missed observations and premature judgements.
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University of South Florida- Phase Change Materials (PCM)
Goswami, Yogi; Stefanakos, Lee
2018-05-30
USF is developing low-cost, high-temperature phase-change materials (PCMs) for use in thermal energy storage systems. Heat storage materials are critical to the energy storage process. In solar thermal storage systems, heat can be stored in these materials during the day and released at night--when the sun is not out--to drive a turbine and produce electricity. In nuclear storage systems, heat can be stored in these materials at night and released to produce electricity during daytime peak-demand hours. Most PCMs do not conduct heat very well. Using an innovative, electroless encapsulation technique, USF is enhancing the heat transfer capability of its PCMs. The inner walls of the capsules will be lined with a corrosion-resistant, high-infrared emissivity coating, and the absorptivity of the PCM will be controlled with the addition of nano-sized particles. USF's PCMs remain stable at temperatures from 600 to 1,000°C and can be used for solar thermal power storage, nuclear thermal power storage, and other applications.
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Advancing Fire Weather Research via Interagency Collaboration: The NOAA/USFS MOU
NASA Astrophysics Data System (ADS)
Schranz, S.; Pouyat, R.
2012-12-01
In 2005, the Western Governors' Association (WGA) first articulated the need for closer collaboration between NOAA and the land management agencies to improve our services - and to ensure the best new technology and scientific advances are infused into fire weather information and services. NOAA has taken the WGA advice very seriously and, over the past few years, have followed up by polling users of our fire weather information. This was done both by our Office of the Federal Coordinator for Meteorology, and via an examination of internal and collaborative research activities as conducted by NOAA's Science Advisory Board. Through these processes, and given the tight budget environment, it's become clear we can't make needed progress alone. We need to call upon our joint expertise, along with the expertise of partners across the federal, state, academic, and research communities. This talk will outline the NOAA/USFS MOU signed in August, 2012 and the collaborative research already begun with the USFS and other partners.
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Cheng, Bingbing; Bandi, Venugopal; Wei, Ming-Yuan; Pei, Yanbo; D’Souza, Francis; Nguyen, Kytai T.; Hong, Yi; Yuan, Baohong
2016-01-01
For many years, investigators have sought after high-resolution fluorescence imaging in centimeter-deep tissue because many interesting in vivo phenomena—such as the presence of immune system cells, tumor angiogenesis, and metastasis—may be located deep in tissue. Previously, we developed a new imaging technique to achieve high spatial resolution in sub-centimeter deep tissue phantoms named continuous-wave ultrasound-switchable fluorescence (CW-USF). The principle is to use a focused ultrasound wave to externally and locally switch on and off the fluorophore emission from a small volume (close to ultrasound focal volume). By making improvements in three aspects of this technique: excellent near-infrared USF contrast agents, a sensitive frequency-domain USF imaging system, and an effective signal processing algorithm, for the first time this study has achieved high spatial resolution (~ 900 μm) in 3-centimeter-deep tissue phantoms with high signal-to-noise ratio (SNR) and high sensitivity (3.4 picomoles of fluorophore in a volume of 68 nanoliters can be detected). We have achieved these results in both tissue-mimic phantoms and porcine muscle tissues. We have also demonstrated multi-color USF to image and distinguish two fluorophores with different wavelengths, which might be very useful for simultaneously imaging of multiple targets and observing their interactions in the future. This work has opened the door for future studies of high-resolution centimeter-deep tissue fluorescence imaging. PMID:27829050
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Cheng, Bingbing; Bandi, Venugopal; Wei, Ming-Yuan; Pei, Yanbo; D'Souza, Francis; Nguyen, Kytai T; Hong, Yi; Yuan, Baohong
2016-01-01
For many years, investigators have sought after high-resolution fluorescence imaging in centimeter-deep tissue because many interesting in vivo phenomena-such as the presence of immune system cells, tumor angiogenesis, and metastasis-may be located deep in tissue. Previously, we developed a new imaging technique to achieve high spatial resolution in sub-centimeter deep tissue phantoms named continuous-wave ultrasound-switchable fluorescence (CW-USF). The principle is to use a focused ultrasound wave to externally and locally switch on and off the fluorophore emission from a small volume (close to ultrasound focal volume). By making improvements in three aspects of this technique: excellent near-infrared USF contrast agents, a sensitive frequency-domain USF imaging system, and an effective signal processing algorithm, for the first time this study has achieved high spatial resolution (~ 900 μm) in 3-centimeter-deep tissue phantoms with high signal-to-noise ratio (SNR) and high sensitivity (3.4 picomoles of fluorophore in a volume of 68 nanoliters can be detected). We have achieved these results in both tissue-mimic phantoms and porcine muscle tissues. We have also demonstrated multi-color USF to image and distinguish two fluorophores with different wavelengths, which might be very useful for simultaneously imaging of multiple targets and observing their interactions in the future. This work has opened the door for future studies of high-resolution centimeter-deep tissue fluorescence imaging.
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Robbins, Lisa L.; Knorr, Paul O.; Daly, Kendra L.; Taylor, Carl A.
2014-01-01
During January and February 2011 the U.S. Geological Survey (USGS), in cooperation with the University of South Florida (USF), conducted geochemical surveys on the west Florida Shelf. Data collected will allow USGS and USF scientists to investigate the effects of climate change on ocean acidification within the northern Gulf of Mexico, specifically, the effect of ocean acidification on marine organisms and habitats. This work is part of a larger USGS study on Climate and Environmental Variability (CEV). The first cruise was conducted from January 3 – 7 (11CEV01) and the second from February 17 - 27 (11CEV02). To view each cruise's survey lines, please see the Trackline page. Both cruises took place aboard the R/V Weatherbird II, a ship of opportunity led by Dr. Kendra Daly (USF), which departed and returned from Saint Petersburg, Florida. Data collection included sampling of the surface and water column (referred to as station samples) with lab analysis of pH, dissolved inorganic carbon (DIC), and total alkalinity. Augmenting the lab analysis was a continuous flow-through system with a Conductivity-Temperature-Depth (CTD) sensor, which also recorded salinity, and pH. Corroborating the USGS data are the vertical CTD profiles collected by USF. The CTD casts measured continuous vertical profiles of oxygen, chlorophyll fluorescence, optical backscatter, and transmissometer. Discrete samples for nutrients, chlorophyll, and particulate organic carbon/nitrogen were also collected during the CTD casts.
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Library and Information Skills Workbook.
ERIC Educational Resources Information Center
Vastine, Jim
This workbook for a library and information skills course at the University of South Florida (USF), Tampa campus, contains the following sections: (1) syllabus; (2) tentative course outline; (3) statement of the course goal, general objectives, and objectives related to LC (Library of Congress) classification and the online catalog, dictionaries…
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Nakajima, Isao
2010-12-01
Due to the socioeconomic reason in Japan, some cancer patient is sometimes operated at a rural hospital where only several surgeons perform and no pathologist checks its malignancy. Therefore, the system of the remote diagnosis for frozen section has been standing up in this country for 7 years. In Japan, the USF has started from February 2007 to support only telecommunications operator's hardware (NTT's equipment such as digital switch board) in high cost areas, not for the reimbursement of the tariff of the public users, such as telepathology. To solve such social cormorant equality, when the USF and PAs were supported in the present quick frozen intraoperative telepathology diagnosis, the quality of the cancer treatment in rural area will be improved. Based on the past data of the Japanese telepathology with beta distribution function, it can be estimated that user terminals becomes five times more than present users with support of USF and PAs. Moreover, using VPN on the B'FLETS, the effect of other teleconsultations will spread to the nationwide.
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Biogeographic, cultural, and historical setting [Chapter 2
Hanna K. Olson; Don W. Fallon
2018-01-01
The Intermountain Adaptation Partnership (IAP) encompasses unique landscapes within the Intermountain Region of the U.S. Forest Service (USFS), from rugged mountains to deep canyons, from alpine snowfields to wild and scenic rivers (fig. 1.1). The area defined by the boundaries of the Intermountain Region contains both private and Federally owned lands, including 12...
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Mark S. Riedel; James M. Vose
2003-01-01
The Coweeta Hydrologic Laboratory, USFS Southern Research Station, worked with state and local agencies and various organizations to provide guidance and tools to reduce sedimentation and facilitate restoration of the 1900km2 Conasauga River watershed in northern Georgia and southern Tennessee. The Conasauga River has the most diverse aquatic...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-22
... Competition Bureau Seeks Updates and Corrections to TelcoMaster Table for Connect America Cost Model AGENCY... centers to particular holding companies for purposes of Connect America Phase II implementation. DATES... companies for purposes of Connect America Phase II implementation. 2. The USF/ICC Transformation Order, 76...
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The sensitivity of relative toxicity rankings by the USF/NASA test method to some test variables
NASA Technical Reports Server (NTRS)
Hilado, C. J.; Labossiere, L. A.; Leon, H. A.; Kourtides, D. A.; Parker, J. A.; Hsu, M.-T. S.
1976-01-01
Pyrolysis temperature and the distance between the source and sensor of effluents are two important variables in tests for relative toxicity. Modifications of the USF/NASA toxicity screening test method to increase the upper temperature limit of pyrolysis, reduce the distance between the sample and the test animals, and increase the chamber volume available for animal occupancy, did not significantly alter rankings of relative toxicity of four representative materials. The changes rendered some differences no longer significant, but did not reverse any rankings. The materials studied were cotton, wool, aromatic polyamide, and polybenzimidazole.
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Christensen, Lise Lotte; Tobiasen, Heidi; Holm, Anja; Schepeler, Troels; Ostenfeld, Marie S; Thorsen, Kasper; Rasmussen, Mads H; Birkenkamp-Demtroeder, Karin; Sieber, Oliver M; Gibbs, Peter; Lubinski, Jan; Lamy, Philippe; Laurberg, Søren; Oster, Bodil; Hansen, Kristian Q; Hagemann-Madsen, Rikke; Byskov, Kristina; Ørntoft, Torben F; Andersen, Claus L
2013-07-01
Colorectal cancer (CRC) is one of the leading causes of cancer deaths in Western countries. A significant number of CRC patients undergoing curatively intended surgery subsequently develop recurrence and die from the disease. MicroRNAs (miRNAs) are aberrantly expressed in cancers and appear to have both diagnostic and prognostic significance. In this study, we identified novel miRNAs associated with recurrence of CRC, and their possible mechanism of action. TaqMan(®) Human MicroRNA Array Set v2.0 was used to profile the expression of 667 miRNAs in 14 normal colon mucosas and 46 microsatellite stable CRC tumors. Four miRNAs (miR-362-3p, miR-570, miR-148 a* and miR-944) were expressed at a higher level in tumors from patients with no recurrence (p<0.015), compared with tumors from patients with recurrence. A significant association with increased disease free survival was confirmed for miR-362-3p in a second independent cohort of 43 CRC patients, using single TaqMan(®) microRNA assays. In vitro functional analysis showed that over-expression of miR-362-3p in colon cancer cell lines reduced cell viability, and proliferation mainly due to cell cycle arrest. E2F1, USF2 and PTPN1 were identified as potential miR-362-3p targets by mRNA profiling of HCT116 cells over-expressing miR-362-3p. Subsequently, these genes were confirmed as direct targets by Luciferase reporter assays and their knockdown in vitro phenocopied the effects of miR-362-3p over-expression. We conclude that miR-362-3p may be a novel prognostic marker in CRC, and hypothesize that the positive effects of augmented miR-362-3p expression may in part be mediated through the targets E2F1, USF2 and PTPN1. Copyright © 2012 UICC.
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Temperature-dependent stability of stacking faults in Al, Cu and Ni: first-principles analysis.
Bhogra, Meha; Ramamurty, U; Waghmare, Umesh V
2014-09-24
We present comparative analysis of microscopic mechanisms relevant to plastic deformation of the face-centered cubic (FCC) metals Al, Cu, and Ni, through determination of the temperature-dependent free energies of intrinsic and unstable stacking faults along [1 1̄ 0] and [1 2̄ 1] on the (1 1 1) plane using first-principles density-functional-theory-based calculations. We show that vibrational contribution results in significant decrease in the free energy of barriers and intrinsic stacking faults (ISFs) of Al, Cu, and Ni with temperature, confirming an important role of thermal fluctuations in the stability of stacking faults (SFs) and deformation at elevated temperatures. In contrast to Al and Ni, the vibrational spectrum of the unstable stacking fault (USF[1 2̄ 1]) in Cu reveals structural instabilities, indicating that the energy barrier (γusf) along the (1 1 1)[1 2̄ 1] slip system in Cu, determined by typical first-principles calculations, is an overestimate, and its commonly used interpretation as the energy release rate needed for dislocation nucleation, as proposed by Rice (1992 J. Mech. Phys. Solids 40 239), should be taken with caution.
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Development of "Remotely Operated Vehicles for Education and Research" (ROVERs)
NASA Astrophysics Data System (ADS)
Gaines, J. E.; Bland, G.; Bydlowski, D.
2017-12-01
The University of South Florida is a team member for the AREN project which develops educational technologies for data acquisition. "Remotely Operated Vehicles for Education and Research" (ROVERs) are floatable data acquisition systems used for Earth science measurements. The USF partnership was productive in the first year, resulting in new autonomous ROVER platforms being developed and used during a 5 week STEM summer camp by middle school youth. ROVERs were outfitted with GPS and temperature sensors and programmed to move forward, backwards, and to turn autonomously using the National Instruments myRIO embedded system. GLOBE protocols were used to collect data. The outreach program's structure lended itself to accomplishing an essential development effort for the AREN project towards the use of the ROVER platform in informal educational settings. A primary objective of the partnership is curriculum development to integrate GLOBE protocols and NASA technology and hardware/ROVER development wher new ROVER platforms are explored. The USF partnership resulted in two design prototypes for ROVERs, both of which can be created from recyclable materials for flotation and either 3D printed or laser cut components. In addition, both use the National Instruments myRIO for autonomous control. We will present two prototypes designed for use during the USF outreach program, the structure of the program, and details on the fabrication of prototype Z during the program by middle school students. Considering the 5-year objective of the AREN project is to "develop approaches, learning plans, and specific tools that can be affordably implemented nationwide (globally)", the USF partnership is key as it contributes to each part of the objective in a unique and impactful way.
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2010-01-30
Kickback Act and making false statements 7/20/2007 14 months in prison; 2 years supervised release; $6,000 fine; $17,964 restitution Anthony Martin ...usf-iraq.com/news/press-briefings/generals-david- petrae - us-and-ray-odierno-take-media-questions-following-the- establishment-of-united-states-forces
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USF Sarasota-Manatee Work Plan Presentation for 2014-15 Board of Governors Review
ERIC Educational Resources Information Center
Board of Governors, State University System of Florida, 2014
2014-01-01
The State University System of Florida has developed three tools that aid in guiding the System's future: (1) The Board of Governors' new "Strategic Plan 2012-2025" is driven by goals and associated metrics that stake out where the System is headed; (2) The Board's "Annual Accountability Report" provides yearly tracking for how…
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75 FR 21036 - Notice of Proposed Withdrawal Extension and Opportunity for Public Meeting; Oregon
Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-22
...,400 acres of National Forest System land from mining in order to protect the major anadromous fish...: Bureau of Land Management, Interior. ACTION: Notice. SUMMARY: The United States Forest Service (USFS) has..., from location and entry under the United States mining laws (30 U.S.C. ch. 2), for an additional 20...
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The Stephen F. Austin Experimental Forest
Cary C. Russell; Ronald E. Thill; David L. Kulhavy
2002-01-01
On December 14, 1944, the Seventy-Eighth United States Congress passed a bill that authorized the transfer of 2,560 acres in Nacogdoches County, Texas, to the research branch of the United States Forest Service (USFS). This land became the Stephen F. Austin Experimental Forest (SFAEF) on September 19. 1945. One of eighty-one federal experimental forests and ranges...
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NASA Astrophysics Data System (ADS)
Mayo, M.; Williams, C.; Rodriguez, T.; Greely, T.; Pyrtle, A. J.; Rivera-Rentas, A. L.; Vilches, M.
2004-12-01
The National Science Foundation's Graduate Teaching Fellows in K-12 Education (GK-12) Program has enabled science, technology, engineering and mathematics (STEM) graduate schools across the country to become more active in local area K-12 schools. An overview of a graduate student's experiences, insights gained and lessons learned as a Fellow in the 2003-2004 Universidad Metropolitana's (UMET) environmental science and the 2004-2005 University of South Florida's (USF) ocean science GK-12 Programs is presented. The major goals of the 2003-2004 UMET GK-12 Program were 1) to enrich environmental science teaching and learning via a thematic approach in eight local public schools and 2) to provide UMET graduate students with exposure to teaching methodologies and practical teaching experience. Utilizing examples from local environments in and nearby Carolina, Puerto Rico to teach key science principles at Escuela de la Comunidad Juana Rodriguez Mundo provided numerous opportunities to relate science topics to students' daily life experiences. By 2004, the UMET GK-12 Program had successfully engaged the entire student body (primarily comprised of bilingual minority kindergarten to sixth graders), teachers and school administrators in environment-focused teaching and learning activities. Examples of such activities include tree planting projects to minimize local erosion, conducting a science fair for the first time in many years, and numerous opportunities to experience what "real scientists do" while conducting environmental science investigations. During the 2004-2005 academic year, skills, insights and lessons learned as a UMET GK-12 Fellow are being further enhanced through participation in the USF GK-12 OCEANS Program. The overall objectives of the 2004-2005 USF GK-12 OCEANS assignment at Madeira Beach Elementary School in Saint Petersburg, Florida are to 1) engage students from various ethnic backgrounds and cultures in hands-on science activities, 2) enhance the school's third grade ocean science education curriculum, and 3) foster dialog between students at Madeira Beach Elementary School and Escuela de la Comunidad Juana Rodriguez Mundo, via exchange of pictures, video recordings, letters and emails related to environment-focused learning activities being undertaken at the two schools. In addition to these objectives, during the 2004-2005 academic year several ocean science-focused activities, the majority of which were adapted and/or identified from either the UMET GK-12 or USF OCEAN GK-12 Programs, will be utilized to further stimulate Madeira Beach Elementary School third graders' critical thinking skills. Examples of such activities, including hands-on exercises, case studies, games and field trips are highlighted in this presentation.
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Copper-silver deposits of the Revett Formation, Montana and Idaho: origin and resource potential
Frost, Thomas P.; Zientek, Michael L.
2006-01-01
The Revett Formation of northern Idaho and western Montana contains major stratabound copper-silver deposits near Troy, Rock Creek, and Rock Lake, Montana. To help the U.S. Forest Service (USFS) meet its goal of integrating geoscience information into the land-planning process, U.S. Geological Survey (USGS) scientists recently completed a compilation of regional stratigraphy and mineralogy of the Revett Formation and a mineral resource assessment of Revett-type copper-silver deposits. The USGS assessment indicates that a large area of USFS-administered land in northwestern Montana and northern Idaho may contain significant undiscovered Revett-type copper-silver deposits.
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Dery, Kenneth J; Silver, Craig; Yang, Lu; Shively, John E
2018-06-15
The adhesion protein carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is widely expressed in epithelial cells as a short cytoplasmic isoform (S-iso) and in leukocytes as a long cytoplasmic isoform (L-iso) and is frequently silenced in cancer by unknown mechanisms. Previously, we reported that interferon response factor 1 (IRF1) biases alternative splicing (AS) to include the variable exon 7 (E7) in CEACAM1, generating long cytoplasmic isoforms. We now show that IRF1 and a variant of heterogeneous nuclear ribonucleoprotein L (Lv1) coordinately silence the CEACAM1 gene. RNAi-mediated Lv1 depletion in IRF1-treated HeLa and melanoma cells induced significant CEACAM1 protein expression, reversed by ectopic Lv1 expression. The Lv1-mediated CEACAM1 repression resided in residues Gly 71 -Gly 89 and Ala 38 -Gly 89 in Lv1's N-terminal extension. ChIP analysis of IRF1- and FLAG-tagged Lv1-treated HeLa cells and global treatment with the global epigenetic modifiers 5-aza-2'-deoxycytidine and trichostatin A indicated that IRF1 and Lv1 together induce chromatin remodeling, restricting IRF1 access to the CEACAM1 promoter. In interferon γ-treated HeLa cells, the transcription factor SP1 did not associate with the CEACAM1 promoter, but binding by upstream transcription factor 1 (USF1), a known CEACAM1 regulator, was greatly enhanced. ChIP-sequencing revealed that Lv1 overexpression in IRF1-treated cells induces transcriptional silencing across many genes, including DCC ( d eleted in c olorectal c arcinoma), associated with CEACAM5 in colon cancer. Notably, IRF1, but not IRF3 and IRF7, affected CEACAM1 expression via translational repression. We conclude that IRF1 and Lv1 coordinately regulate CEACAM1 transcription, alternative splicing, and translation and may significantly contribute to CEACAM1 silencing in cancer. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.
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Nelson, Adam C.; Cauceglia, Joseph W.; Merkley, Seth D.; Youngson, Neil A.; Oler, Andrew J.; Nelson, Randy J.; Cairns, Bradley R.; Whitelaw, Emma; Potts, Wayne K.
2013-01-01
When brought into captivity, wild animals can adapt to domestication within 10 generations. Such adaptations may decrease fitness in natural conditions. Many selective pressures are disrupted in captivity, including social behavioral networks. Although lack of sociality in captivity appears to mediate domestication, the underlying mechanisms are not well understood. Additionally, determining the contribution of genetic inheritance vs. transgenerational effects during relaxed selection may provide insight into the flexibility of adaptation. When wild-derived mice kept under laboratory conditions for eight generations were reintroduced to sociality and promiscuity (free mate choice), they adapted within two generations. Fitness assessments between this promiscuous lineage and a monogamous laboratory lineage revealed male-specific effects. Promiscuous-line males had deficits in viability, but a striking advantage in attracting mates, and their scent marks were also more attractive to females. Here, we investigate mechanistic details underlying this olfactory signal and identify a role of major urinary protein (MUP) pheromones. Promiscuous-line males inherit higher MUP expression than monogamous-line males through transgenerational inheritance. Sociality-driven maternal and paternal effects reveal intriguing conflicts among parents and offspring over pheromone expression. MUP up-regulation is not driven by hormone-driven transduction pathways, but rather is associated with reduction in DNA methylation of a CpG dinucleotide in the promoter. This reduction in methylation could enhance transcription by promoting the binding of transcription factor USF1 (upstream stimulatory factor 1). Finally, we experimentally demonstrate that increased MUP expression is a female attractant. These results identify molecular mechanisms guiding domestication and adaptive responses to fluctuating sociality. PMID:24248373
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Test of the Semi-Analytical Case 1 and Gelbstoff Case 2 SeaWiFS Algorithm with a Global Data Set
NASA Technical Reports Server (NTRS)
Carder, Kendall L.
1997-01-01
The algorithm-development activities at USF during the second half of 1997 have concentrated on data collection and theoretical modeling. Six abstracts were submitted for presentation at the AGU conference in San Diego, California during February 9-13, 1998. Four papers were submitted to JGR and Applied Optics for publication.
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Yuan, Baohong; Pei, Yanbo; Kandukuri, Jayanth
2013-01-01
Our recently developed ultrasound-switchable fluorescence (USF) imaging technique showed that it was feasible to conduct high-resolution fluorescence imaging in a centimeter-deep turbid medium. Because the spatial resolution of this technique highly depends on the ultrasound-induced temperature focal size (UTFS), minimization of UTFS becomes important for further improving the spatial resolution USF technique. In this study, we found that UTFS can be significantly reduced below the diffraction-limited acoustic intensity focal size via nonlinear acoustic effects and thermal confinement by appropriately controlling ultrasound power and exposure time, which can be potentially used for deep-tissue high-resolution imaging. PMID:23479498
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A genomewide survey of basic helix–loop–helix factors in Drosophila
Moore, Adrian W.; Barbel, Sandra; Jan, Lily Yeh; Jan, Yuh Nung
2000-01-01
The basic helix–loop–helix (bHLH) transcription factors play important roles in the specification of tissue type during the development of animals. We have used the information contained in the recently published genomic sequence of Drosophila melanogaster to identify 12 additional bHLH proteins. By sequence analysis we have assigned these proteins to families defined by Atonal, Hairy-Enhancer of Split, Hand, p48, Mesp, MYC/USF, and the bHLH-Per, Arnt, Sim (PAS) domain. In addition, one single protein represents a unique family of bHLH proteins. mRNA in situ analysis demonstrates that the genes encoding these proteins are expressed in several tissue types but are particularly concentrated in the developing nervous system and mesoderm. PMID:10973473
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Takahashi, T; Guron, C; Shetty, S; Matsui, H; Raghow, R
1997-09-05
To dissect the cis-regulatory elements of the murine Msx-1 promoter, which lacks a conventional TATA element, a putative Msx-1 promoter DNA fragment (from -1282 to +106 base pairs (bp)) or its congeners containing site-specific alterations were fused to luciferase reporter and introduced into NIH3T3 and C2C12 cells, and the expression of luciferase was assessed in transient expression assays. The functional consequences of the sequential 5' deletions of the promotor revealed that multiple positive and negative regulatory elements participate in regulating transcription of the Msx-1 gene. Surprisingly, however, the optimal expression of Msx-1 promoter in either NIH3T3 or C2C12 cells required only 165 bp of the upstream sequence to warrant detailed examination of its structure. Therefore, the functional consequences of site-specific deletions and point mutations of the cis-acting elements of the minimal Msx-1 promoter were systematically examined. Concomitantly, potential transcriptional factor(s) interacting with the cis-acting elements of the minimal promoter were also studied by gel electrophoretic mobility shift assays and DNase I footprinting. Combined analyses of the minimal promoter by DNase I footprinting, electrophoretic mobility shift assays, and super shift assays with specific antibodies revealed that 5'-flanking regions from -161 to -154 and from -26 to -13 of the Msx-1 promoter contains an authentic E box (proximal E box), capable of binding a protein immunologically related to the upstream stimulating factor 1 (USF-1) and a GC-rich sequence motif which can bind to Sp1 (proximal Sp1), respectively. Additionally, we observed that the promoter activation was seriously hampered if the proximal E box was removed or mutated, and the promoter activity was eliminated completely if the proximal Sp1 site was similarly altered. Absolute dependence of the Msx-1 minimal promoter on Sp1 could be demonstrated by transient expression assays in the Sp1-deficient Drosophila cell line cotransfected with Msx-1-luciferase and an Sp1 expression vector pPacSp1. The transgenic mice embryos containing -165/106-bp Msx-1 promoter-LacZ DNA in their genomes abundantly expressed beta-galactosidase in maxillae and mandibles and in the cellular primordia involved in the formation of the meninges and the bones of the skull. Thus, the truncated murine Msx-1 promoter can target expression of a heterologous gene in the craniofacial tissues of transgenic embryos known for high level of expression of the endogenous Msx-1 gene and found to be severely defective in the Msx-1 knock-out mice.
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Collaboration in Action: Office of Research and Development ...
The "Collaboration in Action: US EPA's Office of Research and Develop - Current Wildfire Research Program" was invited by the USDA's US Forest Service's Scientific Executive Committee to provide USFS scientific leadership active and potential future opportunities for cooperation/collaboration. Health impacts of wildfire smoke merit the attention and action of the US EPA and current research is supported in the ACE and SHC Research Programs. Wildland fire smoke research has taken on greater importance because the 1) contribution of wildland fire PM emissions relative to total US PM emissions is increasing, 2) the population health impacts are measurable and costly, 3) vulnerable and sensitive populations at-risk are increasing attendant to our aging U.S. population and the increasing area of the wildland-urban interface, and 4) health impacts of smoke could be minimized by identifying at-risk individuals and reducing their exposures. Examples are provided. The "Collaboration in Action: US EPA's Office of Research and Develop - Current Wildfire Research Program" was invited by the USDA's US Forest Service's Scientific Executive Committee to provide USFS scientific leadership active and potential future opportunities for cooperation/collaboration.
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Carlson, Mary H.; Zientek, Michael L.; Causey, J. Douglas; Kayser, Helen Z.; Spanski, Gregory T.; Wilson, Anna B.; Van Gosen, Bradley S.; Trautwein, Charles M.
2007-01-01
This report compiles selected results from 13 U.S. Geological Survey (USGS) mineral resource assessment studies conducted in Idaho and Montana into consistent spatial databases that can be used in a geographic information system. The 183 spatial databases represent areas of mineral potential delineated in these studies and include attributes on mineral deposit type, level of mineral potential, certainty, and a reference. The assessments were conducted for five 1? x 2? quadrangles (Butte, Challis, Choteau, Dillon, and Wallace), several U.S. Forest Service (USFS) National Forests (including Challis, Custer, Gallatin, Helena, and Payette), and one Bureau of Land Management (BLM) Resource Area (Dillon). The data contained in the spatial databases are based on published information: no new interpretations are made. This digital compilation is part of an ongoing effort to provide mineral resource information formatted for use in spatial analysis. In particular, this is one of several reports prepared to address USFS needs for science information as forest management plans are revised in the Northern Rocky Mountains.
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Three-dimensional dictionary-learning reconstruction of (23)Na MRI data.
Behl, Nicolas G R; Gnahm, Christine; Bachert, Peter; Ladd, Mark E; Nagel, Armin M
2016-04-01
To reduce noise and artifacts in (23)Na MRI with a Compressed Sensing reconstruction and a learned dictionary as sparsifying transform. A three-dimensional dictionary-learning compressed sensing reconstruction algorithm (3D-DLCS) for the reconstruction of undersampled 3D radial (23)Na data is presented. The dictionary used as the sparsifying transform is learned with a K-singular-value-decomposition (K-SVD) algorithm. The reconstruction parameters are optimized on simulated data, and the quality of the reconstructions is assessed with peak signal-to-noise ratio (PSNR) and structural similarity (SSIM). The performance of the algorithm is evaluated in phantom and in vivo (23)Na MRI data of seven volunteers and compared with nonuniform fast Fourier transform (NUFFT) and other Compressed Sensing reconstructions. The reconstructions of simulated data have maximal PSNR and SSIM for an undersampling factor (USF) of 10 with numbers of averages equal to the USF. For 10-fold undersampling, the PSNR is increased by 5.1 dB compared with the NUFFT reconstruction, and the SSIM by 24%. These results are confirmed by phantom and in vivo (23)Na measurements in the volunteers that show markedly reduced noise and undersampling artifacts in the case of 3D-DLCS reconstructions. The 3D-DLCS algorithm enables precise reconstruction of undersampled (23)Na MRI data with markedly reduced noise and artifact levels compared with NUFFT reconstruction. Small structures are well preserved. © 2015 Wiley Periodicals, Inc.
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Earth Science Capability Demonstration Project
NASA Technical Reports Server (NTRS)
Cobleigh, Brent
2006-01-01
A viewgraph presentation reviewing the Earth Science Capability Demonstration Project is shown. The contents include: 1) ESCD Project; 2) Available Flight Assets; 3) Ikhana Procurement; 4) GCS Layout; 5) Baseline Predator B Architecture; 6) Ikhana Architecture; 7) UAV Capability Assessment; 8) The Big Picture; 9) NASA/NOAA UAV Demo (5/05 to 9/05); 10) NASA/USFS Western States Fire Mission (8/06); and 11) Suborbital Telepresence.
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Keith D. Stockmann; Nathaniel M. Anderson; Kenneth E. Skog; Sean P. Healey; Dan R. Loeffler; Greg Jones; James F. Morrison
2012-01-01
Global forests capture and store significant amounts of CO2 through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Characterization of BRCA2 Transcriptional Regulation
2001-12-01
tig of BRCA2 promoter construct and 0.1 and we verify the role of USF in regulation of basal activity of jig of pRL-TK Renilla luciferase vector...Promega) with 4 1 l of Fugene-6 the promoter. was used for each transfection. The pRL-TK Renilla luciferase activity was used to control for transfection...pCMV-CREB, pCMV-Myc, BRCA2 Reporter Constructs-A BAC clone (B489G) containing the 5’ and pCMV-Max. Firefly luciferase and Renilla luciferase assays
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Lee, J S; Kim, J M; Hong, E K; Kim, S-O; Yoo, Y-J; Cha, J-H
2009-02-01
A growing amount of attention has been placed on periodontal regeneration and wound healing for periodontal therapy. This study was conducted in an effort to determine the effects of heparin-binding epidermal growth factor-like growth factor on cell repopulation and signal transduction in periodontal ligament cells after scratch wounding in vitro. Human periodontal ligament cells were acquired from explant tissue of human healthy periodontal ligament. After the wounding of periodontal ligament cells, the change in expression of heparin-binding epidermal growth factor-like growth factor and epidermal growth factor receptors 1-4 mRNA was assessed. The effects of heparin-binding epidermal growth factor-like growth factor on periodontal ligament cell proliferation and repopulation were assessed in vitro via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and by photographing the injuries, respectively. Extracellular signal-regulated kinase (Erk)1/2, p38 and Akt phosphorylation was characterized via western blotting. Scratch wounding resulted in a significant up-regulation of heparin-binding epidermal growth factor-like growth factor mRNA expression, whereas wounding had no effect on the expression levels of epidermal growth factor receptors 1-4. Interestingly, no expression of epidermal growth factor receptors 2 and 4 was detectable prior to or after wounding. Heparin-binding epidermal growth factor-like growth factor treatment promoted the proliferation and repopulation of periodontal ligament cells. The scratch wounding also stimulated the phosphorylation of Erk1/2 and p38, but not of Akt, in periodontal ligament cells, and heparin-binding epidermal growth factor-like growth factor treatment applied after wounding amplified and extended the activations of Erk1/2 and p38, but not of Akt. Furthermore, Erk1/2 inhibition blocked the process of cell repopulation induced by heparin-binding epidermal growth factor-like growth factor, whereas the inhibition of p38 delayed the process. These results indicate that heparin-binding epidermal growth factor-like growth factor may constitute a critical factor in the wound healing of human periodontal ligament cells by a mechanism that requires the activation of Erk1/2 via specific interaction with epidermal growth factor receptor 1.
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[Linkage analysis of a family with familial hypertriglyceridemia].
Tang, Xin; Lin, Ying; Liu, Bing; Ma, Shi; Yang, Yang; Yang, Zheng-lin
2009-10-01
To perform linkage analysis and mutation screening in a Chinese family with familial hpertriglyceridemia (FHTG). Thirty-two family members including 12 hypertriglyceridemia patients participated in the study. Genotyping and haplotype analysis for 22 subjects were performed using short tandem repeat (STR) microsatellite polymorphism markers on 16 candidate genes and/or loci related to lipid metabolism. Two of the sixteen known candidate genes, APOA2 and USF1 were screened for mutation by direct DNA sequencing. No linkage was found between the candidate genes/loci of APOA5, LIPI, RP1, APOC2, ABC1, LMF1, APOA1-APOC3-APOA4, LPL, APOB, CETP, LCAT, LDLR, APOE and the phenotype in this family. The two-point Lod scores (theta =0) were all less than-1.0 for all the markers tested. Linkage analysis suggested linkage to chromosome 1q23.3-24.2 between the disease phenotype and STR marker D1S194 with a two-point maximum Lod score of 2.44 at theta =0. Fine mapping indicated that the disease gene was localized to a 5.87 cM interval between D1S104 and D1S196. No disease-causing mutation was detected in the APOA2 and USF1 genes. The above mentioned candidate genes were excluded as the disease causing genes for this family. The results implied that there might be a novel gene/locus for FHTG on chromosome 1q23.3-1q24.2.
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ERIC Educational Resources Information Center
Herreid, Charlene H.; Miller, Thomas E.
2009-01-01
This article is the fourth in a series of articles describing an attrition prediction and intervention project at the University of South Florida (USF) in Tampa. In this article, the researchers describe the updated version of the prediction model. The original model was developed from a sample of about 900 First Time in College (FTIC) students…
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N. Anderson; J. Young; K. Stockmann; K. Skog; S. Healey; D. Loeffler; J.G. Jones; J. Morrison
2013-01-01
Global forests capture and store significant amounts of CO2 through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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2016-03-01
Mbah, PhD2*, Ambuj Kumar, MD, MPH3, Kim Sehwan, PhD4*, Ronald Schonwetter, MD5* and Benjamin Djulbegovic, MD, PhD6 1Center for Evidence - Based Medicine , University...of South Florida, Tampa, FL 2USF, Tampa, FL 3University of South Florida, College of Medicine, Center for Evidence Based Medicine , Tampa...4HPC healthcare, Tampa, FL 5HPC Healthcare, Tampa, FL 6Center for Evidence - Based Medicine & Health Outcomes Research, University of South
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... Global Health Security HIV & Tuberculosis Global Health Protection Malaria & Parasitic Diseases Immunization Other Diseases & Threats Travelers' Health ... Organization Strategy Partnerships Funding Latest News War on malaria: USF researchers wage battle against global disease 83 ...
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High-Volume Airborne Fluids Handling Technologies to Fight Wildfires
NASA Technical Reports Server (NTRS)
Dickerson, Mark; Cox, Timothy; Hale, Cliff; Hatton, Rick
2010-01-01
NASA recently partnered with the U.S. Forest Service (USFS) on a project to examine mission suitability and recommend policies and procedures for the use of very large aerial firefighting aircraft such as the Boeing 747 and DC-10 aerial retardant delivery aircraft. The aircraft under study included a 10Tanker DC-10 and an Evergreen B-747. NASA's Dryden Flight Research Center and Ames Research Center worked with the USFS to help determine the safe flight envelope for these Very Large Air Tanker (VLAT) aircraft for the USFS and the Department of the Interior (DOI). This new generation of supertankers includes aircraft like these that have as much as four times the delivery capacity of the previous generation of aerial firefighting aircraft. Dryden performed operational test and evaluation assessments and reported findings and recommendations on these aircraft in cooperation with Ames. The team developed, implemented, and directed an evaluation test plan for use in flight test and in simulation. Ames provided support using pilot-in-the-loop simulations and coordinated simulator models, flight profiles, and data analysis with Dryden. The test plan was designed to evaluate the suitability of VLAT aircraft as a function of mission environment. Based on this analysis, NASA generated interim flight envelope limitations to enhance safety and operational utility in the fire-retardant delivery mission. These recommended flight limitations were adopted by the USFS. The 10Tanker DC-10 has been in use for several years with the California Department of Forestry and Fire Protection(Cal-Fire), but until NASA took on the challenge of reviewing VLAT capabilities and limitations, the USFS was hesitant to add them to the federal wildfire arsenal. The DC-10 delivery system is based on an externally mounted set of tanks and a bomb-bay style set of clamshell doors that are opened in precisely calibrated ways to deliver the amounts and concentrations of retardant called for by the specific wildfire situation. The system was manufactured by Jordan Air of Central Point, OR, and was installed by Victorville Aerospace in Victorville, CA. It can deliver 12,000 gallons (45.4 kL) of retardant in as little as eight seconds. The aircraft can deliver a partial load of retardant and make multiple drops on the same flight, or the entire load can be rapidly delivered in one pass if required for maximum coverage. The Evergreen 747 uses internal tankage and a pressurized delivery system to enable volume and coverage levels that also meet USFS requirements, but enables computer control of flow for desired precision. This system was designed and built by Adaptive Aerospace of Tehachapi, CA and can deliver about 20,000 gallons (75.7 kL) of retardant in approximately ten seconds. The 747 can also make multiple independent drops, or deliver the entire load at once. NASA found that both of these VLAT aircraft are compatible with the wildfire suppression mission when used to supplement other aerial retardant delivery platforms. The major recommendations for deployment that resulted from this study relate to terrain clearance, the type of terrain in the drop area, availability of qualified lead planes to guide the VLAT approach to the drop area, and low-altitude maneuvering limitations. NASA s analysis suggests that with the appropriate flight procedures, these aircraft will provide a powerful set of tools to fight wildfires.
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NASA Astrophysics Data System (ADS)
Grant, G.; Major, J. J.; Lewis, S.
2016-12-01
The 18 May 1980 eruption of Mount St. Helens, Washington, spawned a massive (109 m3) debris avalanche, a violent and extensive pyroclastic density current, lahars, pyroclastic flows, and ashfall. It fundamentally transformed the proximal landscape, and created potential secondary hazards that remain legacies of the eruption over 35 years later. The debris avalanche raised the level of Spirit Lake—a picturesque lake at the foot of the volcano—by 60 m and blocked its outlet. Abruptly, the lake went from charming to menacing, capable of releasing a potentially catastrophic outburst flood (108 m3) that could transform into a massive (109 m3) debris flow if rising lake water breached the blockage. To reduce risk of an uncontrolled breach, and under Presidential emergency declaration, the U.S. Army Corps of Engineers (USACE) bored a 2,590-m-long outlet tunnel through bedrock within the U.S. Forest Service (USFS)-administered Mount St. Helens National Volcanic Monument. Drainage through the tunnel maintains a safe lake level below a geologic contact in the blockage where seepage erosion could result in failure. Although the tunnel has performed its mission for over 30 years, episodic deformation has reduced its outlet capacity, necessitating expensive (>$1 million) repairs and closures which temporarily caused precarious lake rises, and prompted re-examination of the strategy to maintain a safe lake level. Here we discuss how federal researchers (USFS and U.S. Geological Survey) interact with Monument and USFS land managers, USACE, the National Academy of Sciences, and the public at large to develop and evaluate, under Congressional mandate, alternative strategies for reducing the risk of catastrophic flooding. Amidst this nexus of institutions, agendas, and perspectives, set against the backdrop of a rapidly evolving landscape subject to a trio of hazards (eruptions, earthquakes, floods), competing interests, costs, and natural risks must be balanced and managed.
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76 FR 82296 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-30
.... 20110431, Draft EIS, USFS, NV, Geothermal Leasing on the Humboldt-Toiyabe National Forest, To Facilitate the Development and Production of Geothermal Energy, Ely, Austin, Tonopah and Bridgeport Ranger...
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Chirgadze, Y N; Boshkova, E A; Polozov, R V; Sivozhelezov, V S; Dzyabchenko, A V; Kuzminsky, M B; Stepanenko, V A; Ivanov, V V
2018-01-07
The mouse factor Zif268, known also as early growth response protein EGR-1, is a classical representative for the Cys2His2 transcription factor family. It is required for binding the RNA polymerase with operator dsDNA to initialize the transcription process. We have shown that only in this family of total six Zn-finger protein families the Zn complex plays a significant role in the protein-DNA binding. Electrostatic feature of this complex in the binding of factor Zif268 from Mus musculus with operator DNA has been considered. The factor consists of three similar Zn-finger units which bind with triplets of coding DNA. Essential contacts of the factor with the DNA phosphates are formed by three conservative His residues, one in each finger. We describe here the results of calculations of the electrostatic potentials for the Zn-Cys2His2 complex, Zn-finger unit 1, and the whole transcription factor. The potential of Zif268 has a positive area on the factor surface, and it corresponds exactly to the binding sites of each of Zn-finger units. The main part of these areas is determined by conservative His residues, which form contacts with the DNA phosphate groups. Our result shows that the electrostatic positive potential of this histidine residue is enhanced due to the Zn complex. The other contacts of the Zn-finger with DNA are related to nucleotide bases, and they are responsible for the sequence-specific binding with DNA. This result may be extended to all other members of the Cys2His2 transcription factor family.
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Algorithm-development activities
NASA Technical Reports Server (NTRS)
Carder, Kendall L.
1994-01-01
The task of algorithm-development activities at USF continues. The algorithm for determining chlorophyll alpha concentration, (Chl alpha) and gelbstoff absorption coefficient for SeaWiFS and MODIS-N radiance data is our current priority.
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Human corpus luteum: presence of epidermal growth factor receptors and binding characteristics
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ayyagari, R.R.; Khan-Dawood, F.S.
Epidermal growth factor receptors are present in many reproductive tissues but have not been demonstrated in the human corpus luteum. To determine the presence of epidermal growth factor receptors and its binding characteristics, we carried out studies on the plasma cell membrane fraction of seven human corpora lutea (days 16 to 25) of the menstrual cycle. Specific epidermal growth factor receptors were present in human corpus luteum. Insulin, nerve growth factor, and human chorionic gonadotropin did not competitively displace epidermal growth factor binding. The optimal conditions for corpus luteum-epidermal growth factor receptor binding were found to be incubation for 2more » hours at 4 degrees C with 500 micrograms plasma membrane protein and 140 femtomol /sup 125/I-epidermal growth factor per incubate. The number (mean +/- SEM) of epidermal growth factor binding sites was 12.34 +/- 2.99 X 10(-19) mol/micrograms protein; the dissociation constant was 2.26 +/- 0.56 X 10(-9) mol/L; the association constant was 0.59 +/- 0.12 X 10(9) L/mol. In two regressing corpora lutea obtained on days 2 and 3 of the menstrual cycle, there was no detectable specific epidermal growth factor receptor binding activity. Similarly no epidermal growth factor receptor binding activity could be detected in ovarian stromal tissue. Our findings demonstrate that specific receptors for epidermal growth factor are present in the human corpus luteum. The physiologic significance of epidermal growth factor receptors in human corpus luteum is unknown, but epidermal growth factor may be involved in intragonadal regulation of luteal function.« less
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USF Equitable Distribution Act of 2013
Sen. Ayotte, Kelly [R-NH
2013-11-21
Senate - 11/21/2013 Read twice and referred to the Committee on Commerce, Science, and Transportation. (All Actions) Tracker: This bill has the status IntroducedHere are the steps for Status of Legislation:
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75 FR 36386 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-06-25
..., Revised Draft EIS, USFS, 00, Uinta National Forest Oil and Gas Leasing, Implementation, Identify National Forest Systems Lands with Federal Mineral Rights, Wasatch, Utah, Juab, Tooele, and Sanpete Counties, UT...
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[Interaction of human factor X with thromboplastin].
Kiselev, S V; Zubairov, D M; Timarbaev, V N
2003-01-01
The binding of 125I-labeled human factor X to native and papaine-treated tissue tromboplastin in the presence of CaCl2 or EDTA was studied. The Scatchard analysis suggests the existence of high (Kd=l,8 x10(-9) M) and low affinity binding sites on the thromboplastin surface. The removal of Ca2+ reduced affinity of factor X to the high affinity sites. This was accompanied by some increase of their number. Proteolysis by papaine decreased affinity of high affinity sites and caused the increase of their number in the presence of Ca2+. In the absence of Ca2+ the affinity remained unchanged, but the number of sites decreased. At low concentrations of factor X positive cooperativity for high affinity binding sites was observed. It did not depend on the presence of Ca2+. The results indirectly confirm the role of hydrophobic interactons in Ca2+ dependent binding of factor X to thromboplastin and the fact that heterogeneity of this binding is determined by mesophase structure of the thromboplastin phospholipids.
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NASA Astrophysics Data System (ADS)
Wilcox, William Edward, Jr.
1995-01-01
A computer program (LIDAR-PC) and associated atmospheric spectral databases have been developed which accurately simulate the laser remote sensing of the atmosphere and the system performance of a direct-detection Lidar or tunable Differential Absorption Lidar (DIAL) system. This simulation program allows, for the first time, the use of several different large atmospheric spectral databases to be coupled with Lidar parameter simulations on the same computer platform to provide a real-time, interactive, and easy to use design tool for atmospheric Lidar simulation and modeling. LIDAR -PC has been used for a range of different Lidar simulations and compared to experimental Lidar data. In general, the simulations agreed very well with the experimental measurements. In addition, the simulation offered, for the first time, the analysis and comparison of experimental Lidar data to easily determine the range-resolved attenuation coefficient of the atmosphere and the effect of telescope overlap factor. The software and databases operate on an IBM-PC or compatible computer platform, and thus are very useful to the research community for Lidar analysis. The complete Lidar and atmospheric spectral transmission modeling program uses the HITRAN database for high-resolution molecular absorption lines of the atmosphere, the BACKSCAT/LOWTRAN computer databases and models for the effects of aerosol and cloud backscatter and attenuation, and the range-resolved Lidar equation. The program can calculate the Lidar backscattered signal-to-noise for a slant path geometry from space and simulate the effect of high resolution, tunable, single frequency, and moderate line width lasers on the Lidar/DIAL signal. The program was used to model and analyze the experimental Lidar data obtained from several measurements. A fixed wavelength, Ho:YSGG aerosol Lidar (Sugimoto, 1990) developed at USF and a tunable Ho:YSGG DIAL system (Cha, 1991) for measuring atmospheric water vapor at 2.1 μm were analyzed. The simulations agreed very well with the measurements, and also yielded, for the first time, the ability to easily deduce the atmospheric attentuation coefficient, alpha, from the Lidar data. Simulations and analysis of other Lidar measurements included that of a 1.57 mu m OPO aerosol Lidar system developed at USF (Harrell, 1995) and of the NASA LITE (Laser-in-Space Technology Experiment) Lidar recently flown in the Space shuttle. Finally, an extensive series of laboratory experiments were made with the 1.57 μm OPO Lidar system to test calculations of the telescope/laser overlap and the effect of different telescope sizes and designs. The simulations agreed well with the experimental data for the telescope diameter and central obscuration test cases. The LIDAR-PC programs are available on the Internet from the USAF Lidar Home Page Web site, http://www.cas.usf.edu/physics/lidar.html/.
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75 FR 58376 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-09-24
... Seastrand 626-574-5278. EIS No. 20100378, Draft Supplement, USFS, OR, North Fork Burnt River Mining Project.... 20100380, Final EIS, USACE, 00, Sabine-Neches Waterway Channel Improvement Project, Proposed Ocean Dredged...
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78 FR 59677 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2013-09-27
..., Designation of the Atchafalaya River Bar Channel Ocean Dredged Material Disposal Site, Review Period Ends: 10.../2013, Contact: Paul Bradford 406-293-6211. EIS No. 20130282, Final EIS, USFS, WY, Clinker Mining...
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75 FR 72823 - Environmental Impact Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-26
... Register. EIS No. 20100449, Draft EIS, USFS, MT, Stillwater Mining Revised Water Management Plans and BOE... and Reuse, Implementation, Brunswick, ME, Wait Period Ends: 12/27/2010, Contact: Thomas Stephan 215...
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32 CFR 171.4 - Qualifications.
Code of Federal Regulations, 2010 CFR
2010-07-01
... of meeting the terms and conditions of a contract to deliver fire retardant by air. (a) Prior to... the USDA contract. (b) This requirement may not be delegated to the U.S. Forest Service (USFS). ...
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75 FR 33299 - Environmental Impacts Statements; Notice Of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-06-11
...-963-0182. EIS No. 20100215, Final EIS, USFS, CO, Hermosa Park/Mitchell Lakes Land Exchange Project.../2010 to 07/ 26/2010. Dated: June 8, 2010. Ken Mittelholtz, Deputy Director, NEPA Compliance Division...
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Learning about Antiphospholipid Syndrome (APS)
... rarediseasesnetwork.epi.usf.edu] An integrated group of academic medical centers, patient support organizations, and clinical research ... Diseases (4th ed.), Mackay IR, Rose NR, eds. Academic Press/Elsevier Science. 2006. Williams Hematology, 6th ed., ...
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2003 Florida transportation almanac
DOT National Transportation Integrated Search
2003-12-01
This publication is the third edition of the Florida Transportation Almanac, developed and produced by the : Center for Urban Transportation Research (CUTR) at the University of South Florida (USF) in Tampa. It : follows the original publication prod...
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NASA Astrophysics Data System (ADS)
Tebbens, S. F.; Coble, P. G.; Greely, T.
Three educational outreach programs designed for middle school students (grades 6, 7, and 8) by faculty at the University of South Florida (USF) Department of Marine Science are turning kids onto science. The programs are bringing marine science research and its various technologies into the classroom, where students follow up with hands-on activities. Project Oceanography (PO) is an interactive broadcast that exposes students to the concepts and tools of current marine science research. The Oceanography Camp for Girls (OCG) boosts girls' curiosity and interest in science and nature. And teachers become better equipped to present current marine science topics and technology to their students at the Teachers Oceanography Workshop (TOW). All of the programs created by USF are provided at no cost to students or their institutions.
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Combating the stress of residency: one school's approach.
Dabrow, Sharon; Russell, Stephen; Ackley, Karen; Anderson, Eric; Fabri, Peter Jeff
2006-05-01
Residency is a time of stress and turmoil for many residents. The stresses are varied and great, often involving both personal and professional issues. One institutional mechanism that has been shown to help residents cope with stress is the use of residents' wellness, or assistance, programs. The University of South Florida (USF) College of Medicine developed the USF Residency Assistance Program (RAP) in 1997, modeled after business employee assistance programs but tailored to enhance the well-being of residents. The program was developed in an organized, thoughtful manner starting with a Request for Proposals to all local employee assistance programs and the selection of one of these to run the program. The RAP is broad-based, readily available, easily accessible, totally voluntary and confidential, and not reportable to the state board of medicine. It is well integrated into all residency programs and has had excellent acceptance from the administration; information about access to the RAP is available to all residents through multiple venues. The cost is minimal, at only seven cents a day per resident. The authors present data from the eight years the RAP has been operating, including information on program use, referral rates, acceptance, and types of problems encountered. One suicide occurred during this time period, and the RAP provided a significant role in grief counseling. Assistance programs are critical to the well-being of residents. The USF program presents a model that can be used by other programs around the country.
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Finkernagel, Florian; Stiewe, Thorsten; Nist, Andrea; Suske, Guntram
2015-01-01
Transcription factors are grouped into families based on sequence similarity within functional domains, particularly DNA-binding domains. The Specificity proteins Sp1, Sp2 and Sp3 are paradigmatic of closely related transcription factors. They share amino-terminal glutamine-rich regions and a conserved carboxy-terminal zinc finger domain that can bind to GC rich motifs in vitro. All three Sp proteins are ubiquitously expressed; yet they carry out unique functions in vivo raising the question of how specificity is achieved. Crucially, it is unknown whether they bind to distinct genomic sites and, if so, how binding site selection is accomplished. In this study, we have examined the genomic binding patterns of Sp1, Sp2 and Sp3 in mouse embryonic fibroblasts by ChIP-seq. Sp1 and Sp3 essentially occupy the same promoters and localize to GC boxes. The genomic binding pattern of Sp2 is different; Sp2 primarily localizes at CCAAT motifs. Consistently, re-expression of Sp2 and Sp3 mutants in corresponding knockout MEFs revealed strikingly different modes of genomic binding site selection. Most significantly, while the zinc fingers dictate genomic binding of Sp3, they are completely dispensable for binding of Sp2. Instead, the glutamine-rich amino-terminal region is sufficient for recruitment of Sp2 to its target promoters in vivo. We have identified the trimeric histone-fold CCAAT box binding transcription factor Nf-y as the major partner for Sp2-chromatin interaction. Nf-y is critical for recruitment of Sp2 to co-occupied regulatory elements. Equally, Sp2 potentiates binding of Nf-y to shared sites indicating the existence of an extensive Sp2-Nf-y interaction network. Our results unveil strikingly different recruitment mechanisms of Sp1/Sp2/Sp3 transcription factor members uncovering an unexpected layer of complexity in their binding to chromatin in vivo. PMID:25793500
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NASA Astrophysics Data System (ADS)
Jeuck, James A.
This dissertation consists of research projects related to forest land use / land cover (LULC): (1) factors predicting LULC change and (2) methodology to predict particular forest use, or "potential working timberland" (PWT), from current forms of land data. The first project resulted in a published paper, a meta-analysis of 64 econometric models from 47 studies predicting forest land use changes. The response variables, representing some form of forest land change, were organized into four groups: forest conversion to agriculture (F2A), forestland to development (F2D), forestland to non-forested (F2NF) and undeveloped (including forestland) to developed (U2D) land. Over 250 independent econometric variables were identified, from 21 F2A models, 21 F2D models, 12 F2NF models, and 10 U2D models. These variables were organized into a hierarchy of 119 independent variable groups, 15 categories, and 4 econometric drivers suitable for conducting simple vote count statistics. Vote counts were summarized at the independent variable group level and formed into ratios estimating the predictive success of each variable group. Two ratio estimates were developed based on (1) proportion of times independent variables successfully achieved statistical significance (p ≤0.10), and (2) proportion of times independent variables successfully met the original researchers'expectations. In F2D models, popular independent variables such as population, income, and urban proximity often achieved statistical significance. In F2A models, popular independent variables such as forest and agricultural rents and costs, governmental programs, and site quality often achieved statistical significance. In U2D models, successful independent variables included urban rents and costs, zoning issues concerning forestland loss, site quality, urban proximity, population, and income. F2NF models high success variables were found to be agricultural rents, site quality, population, and income. This meta-analysis provides insight into the general success of econometric independent variables for future forest use or cover change research. The second part of this dissertation developed a method for predicting area estimates and spatial distribution of PWT in the US South. This technique determined land use from USFS Forest Inventory and Analysis (FIA) and land cover from the National Land Cover Database (NLCD). Three dependent variable forms (DV Forms) were derived from the FIA data: DV Form 1, timberland, other; DV Form 2, short timberland, tall timberland, agriculture, other; and DV Form 3, short hardwood (HW) timberland, tall HW timberland, short softwood (SW) timberland, tall SW timberland, agriculture, other. The prediction accuracy of each DV Form was investigated using both random forest model and logistic regression model specifications and data optimization techniques. Model verification employing a "leave-group-out" Monte Carlo simulation determined the selection of a stratified version of the random forest model using one-year NLCD observations with an overall accuracy of 0.53-0.94. The lower accuracy side of the range was when predictions were made from an aggregated NLCD land cover class "grass_shrub". The selected model specification was run using 2011 NLCD and the other predictor variables to produce three levels of timberland prediction and probability maps for the US South. Spatial masks removed areas unlikely to be working forests (protected and urbanized lands) resulting in PWT maps. The area of the resulting maps compared well with USFS area estimates and masked PWT maps and had an 8-11% reduction of the USFS timberland estimate for the US South compared to the DV Form. Change analysis of the 2011 NLCD to PWT showed (1) the majority of the short timberland came from NLCD grass_shrub; (2) the majority of NLCD grass_shrub predicted into tall timberland, and (3) NLCD grass_shrub was more strongly associated with timberland in the Coastal Plain. Resulting map products provide practical analytical tools for those interested in studying the area and distribution of PWT in the US South.
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Performance Monitoring of a Nearshore Berm at Ft. Myers Beach, Florida
2013-08-01
prototype designs. Coastal Zone ’93, American Society of Civil Engineers, pp. 2938 -2950. Andrassy, C . J. 1991. Monitoring of a nearshore disposal mound at...ER D C / CH L TR -1 3 -1 1 Performance Monitoring of a Nearshore Berm at Ft. Myers Beach, Florida: Final Report C oa st al a n d H yd...122 Appendix C : USF-CRL Survey Data: morphologic evolution during the first 2 years post construction
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Different historical fire–climate patterns in California
Keeley, Jon E.; Syphard, Alexandra D.
2017-01-01
The relationship between annual variation in area burned and seasonal temperatures and precipitation was investigated for the major climate divisions in California. Historical analyses showed marked differences in fires on montane and foothill landscapes. Based on roughly a century of data, there are five important lessons on fire–climate relationships in California: (1) seasonal variations in temperature appear to have had minimal influence on area burned in the lower elevation, mostly non-forested, landscapes; (2) temperature has been a significant factor in controlling fire activity in higher elevation montane forests, but this varied greatly with season – winter and autumn temperatures showed no significant effect, whereas spring and summer temperatures were important determinants of area burned; (3) current season precipitation has been a strong controller of fire activity in forests, with drier years resulting in greater area burned on most United States Forest Service (USFS) lands in the state, but the effect of current-year precipitation was decidedly less on lower elevation California Department of Forestry and Fire Protection lands; (4) in largely grass-dominated foothills and valleys the magnitude of prior-year rainfall was positively tied to area burned in the following year, and we hypothesise that this is tied to greater fuel volume in the year following high rainfall. In the southern part of the state this effect has become stronger in recent decades and this likely is due to accelerated type conversion from shrubland to grassland in the latter part of the 20th century; (5) the strongest fire–climate models were on USFS lands in the Sierra Nevada Mountains, and these explained 42–52% of the variation in area burned; however, the models changed over time, with winter and spring precipitation being the primary drivers in the first half of the 20th century, but replaced by spring and summer temperatures after 1960.
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Visitor Access and Transportation Guide.
DOT National Transportation Integrated Search
2011-09-30
This visitor access and transportation guide presents opportunities to manage or improve circulation and travel patterns in areas managed by Federal Land Management Agencies (FLMAs), such as the National Park Service (NPS), U.S. Forest Service (USFS)...
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Platelet binding sites for factor VIII in relation to fibrin and phosphatidylserine
Novakovic, Valerie A.; Shi, Jialan; Rasmussen, Jan; Pipe, Steven W.
2015-01-01
Thrombin-stimulated platelets expose very little phosphatidylserine (PS) but express binding sites for factor VIII (fVIII), casting doubt on the role of exposed PS as the determinant of binding sites. We previously reported that fVIII binding sites are increased three- to sixfold when soluble fibrin (SF) binds the αIIbβ3 integrin. This study focuses on the hypothesis that platelet-bound SF is the major source of fVIII binding sites. Less than 10% of fVIII was displaced from thrombin-stimulated platelets by lactadherin, a PS-binding protein, and an fVIII mutant defective in PS-dependent binding retained platelet affinity. Therefore, PS is not the determinant of most binding sites. FVIII bound immobilized SF and paralleled platelet binding in affinity, dependence on separation from von Willebrand factor, and mediation by the C2 domain. SF also enhanced activity of fVIII in the factor Xase complex by two- to fourfold. Monoclonal antibody (mAb) ESH8, against the fVIII C2 domain, inhibited binding of fVIII to SF and platelets but not to PS-containing vesicles. Similarly, mAb ESH4 against the C2 domain, inhibited >90% of platelet-dependent fVIII activity vs 35% of vesicle-supported activity. These results imply that platelet-bound SF is a component of functional fVIII binding sites. PMID:26162408
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Expansion of the Eclipse Digital Signal Processing System.
1982-12-01
8217eOU WIdT TO,. Fig 1 IE.ETZIM U2. E( 11 -4., - IULTIPI.E P * S WI) STPM FILTER (- PAWtfTEP FILE PFILE FILTER FILE: WILE FIEP. LENGTH 55 WINIIM OF WQS...Vg u I k114 2.2 1 .2 I 11 .l111 1.6 MICROCOPY RESOLUTION TEST CHART NA, ONA BURMAU OF SrANDARDS-1963 A b i -I i.i 1s Lt USF w191 UNITED STATES AIR...SIGNAL PROCESSING SYSTI.M I"’ 1 /GI,/V/H 2 D- I6 Gordon R. Alln ist Lt USAF" I . . SECURITY CLASSIFICATION OF THIS PAGE (When Data Entered) READ
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Update on the Caspar Creek watershed study
Peter Cafferata
1987-01-01
Readers of this Newsletter are aware that CDF and the USFS, through its Pacific Southwest Forest and Range Experiment Stationa at Arcata (PSW), are carrying out a long term cooperative watershed experiment in JDSF's Caspar Creek drainage.
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Impacts of dialysis transportation on Florida's coordinated public transportation programs.
DOT National Transportation Integrated Search
2014-04-01
The National Center for Transit Research (NCTR) at the University of South Florida (USF) collected quantitative and qualitative data from Community Transportation Coordinators (CTCs) throughout Florida. An online survey and a series of personal inter...
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Florida public transportation anti-terrorism resource guide
DOT National Transportation Integrated Search
2001-10-01
The Center for Urban Transportation (CUTR) at the University of South Florida (USF) assembled this guide to provide public transit agencies in Florida with information on current resources available to assist them with improving system security and g...
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Robbins, Lisa L.; Knorr, Paul O.; Daly, Kendra L.; Barrera, Kira E.
2014-01-01
As part of the U.S. Geological Survey (USGS) Coastal and Marine Geology Program project "Response of Florida Shelf Ecosystems to Climate Change" and in partnership with Kendra Daly, University of South Florida (USF), data on surface ocean carbonate chemistry were collected on five cruises along transects on the shallow inner west Florida shelf and northern Gulf of Mexico in 2012. Data from the 2011 cruises were also published (Robbins and others., 2013). The data collected allows the USGS, National Oceanic and Atmospheric Administration (NOAA), and USF scientists to map variations in ocean chemistry including carbonate saturation states along designated tracks. The USGS also partners with NOAA and the National Aeronautics and Space Administration (NASA) to model air-sea flux as part of a Gulf of Mexico Carbon Synthesis project led by NASA.
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NASA Technical Reports Server (NTRS)
Hilado, C. J.; Miller, C. M.
1976-01-01
Rankings of relative toxicity can be markedly affected by changes in test variables. Revision of the USF/NASA toxicity screening test procedure to eliminate the connecting tube and supporting floor and incorporate a 1.0 g sample weight, 200 C starting temperature, and 800 C upper limit temperature for pyrolysis, reversed the rankings of flexible polyurethane and polychloroprene foams, not only in relation to each other, but also in relation to cotton and red oak. Much of the change is attributed to reduction of the distance between the sample and the test animals, and reduction of the sample weight charged. Elimination of the connecting tube increased the relative toxicity of the polyurethane foams. The materials tested were flexible polyurethane foam, without and with fire retardant; rigid polyurethane foam with fire retardant; flexible polychloroprene foam; cotton, Douglas fir, red oak, hemlock, hardboard, particle board, polystyrene, and polymethyl methacrylate.
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2009-04-01
Patient Status ABD (%) Ext (%) Vasc (%) Uro (%) GYN (%) Thor (%) HN (%) Neuro (%) Burn (%) Other (%) Total USF (n 178) 6 (2.6) 125 (54.3) 3 (1.3) 0...Ext, extremity; Vasc, vascular; Uro , urological; GYN, gynecologic; Thor, thoracic; HN, head and neck; Neuro, neurologic. Table 8 Age, Sex, and...Shock Trauma Platoon with a similar patient cohort at Los Angeles County trauma center, found that 12.7% of patients treated by the Surgical Shock
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Salasovich, James; LoVullo, David; Kandt, Alicen
This report summarizes results from the energy efficiency, water efficiency, and renewable energy site assessment of the Mendenhall Glacier Visitor Center and site in Juneau, Alaska. The assessment is an American Society of Heating, Refrigerating, and Air-Conditioning Engineers Level 2 audit and meets Energy Independence and Security Act requirements. A team led by the U.S. Department of Energy's National Renewable Energy Laboratory conducted the assessment with U.S. Forest Service personnel August 19-20, 2015, as part of ongoing efforts by USFS to reduce energy and water use.
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2009-04-01
agreement that the National Aeronautics and Space Administration ( NASA ) has in place. However, with close coordination among all users, C-band...land is owned and administered by the U.S. Bureau of Land Management ( BLM ). White Sands National Monument is located to the southwest. WSMR...for the base. Much of Edwards AFB is surrounded by government lands managed by the BLM , U.S. Forest Service (USFS) and State of California. The
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Numerical modeling of laboratory-scale surface-to-crown fire transition
NASA Astrophysics Data System (ADS)
Castle, Drew Clayton
Understanding the conditions leading to the transition of fire spread from a surface fuel to an elevated (crown) fuel is critical to effective fire risk assessment and management. Surface fires that successfully transition to crown fires can be very difficult to suppress, potentially leading to damages in the natural and built environments. This is relevant to chaparral shrub lands which are common throughout parts of the Southwest U.S. and represent a significant part of the wildland urban interface. The ability of the Wildland-Urban Interface Fire Dynamic Simulator (WFDS) to model surface-to-crown fire transition was evaluated through comparison to laboratory experiments. The WFDS model is being developed by the U.S. Forest Service (USFS) and the National Institute of Standards and Technology. The experiments were conducted at the USFS Forest Fire Laboratory in Riverside, California. The experiments measured the ignition of chamise (Adenostoma fasciculatum) crown fuel held above a surface fire spreading through excelsior fuel. Cases with different crown fuel bulk densities, crown fuel base heights, and imposed wind speeds were considered. Cold-flow simulations yielded wind speed profiles that closely matched the experimental measurements. Next, fire simulations with only the surface fuel were conducted to verify the rate of spread while factors such as substrate properties were varied. Finally, simulations with both a surface fuel and a crown fuel were completed. Examination of specific surface fire characteristics (rate of spread, flame angle, etc.) and the corresponding experimental surface fire behavior provided a basis for comparison of the factors most responsible for transition from a surface fire to the raised fuel ignition. The rate of spread was determined by tracking the flame in the Smokeview animations using a tool developed for tracking an actual flame in a video. WFDS simulations produced results in both surface fire spread and raised fuel bed ignition which closely matched the trends reported in the laboratory experiments.
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Karttunen, Mikko; Choy, Wing-Yiu; Cino, Elio A
2018-06-07
Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor and principal regulator of the antioxidant pathway. The Kelch domain of Kelch-like ECH-associated protein 1 (Keap1) binds to motifs in the N-terminal region of Nrf2, promoting its degradation. There is interest in developing ligands that can compete with Nrf2 for binding to Kelch, thereby activating its transcriptional activities and increasing antioxidant levels. Using experimental Δ G bind values of Kelch-binding motifs determined previously, a revised hydrophobicity-based model was developed for estimating Δ G bind from amino acid sequence and applied to rank potential uncharacterized Kelch-binding motifs identified from interaction databases and BLAST searches. Model predictions and molecular dynamics (MD) simulations suggested that full-length MAD2A binds Kelch more favorably than a high-affinity 20-mer Nrf2 E78P peptide, but that the motif in isolation is not a particularly strong binder. Endeavoring to develop shorter peptides for activating Nrf2, new designs were created based on the E78P peptide, some of which showed considerable propensity to form binding-competent structures in MD, and were predicted to interact with Kelch more favorably than the E78P peptide. The peptides could be promising new ligands for enhancing the oxidative stress response.
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Travel to Food : Transportation Barriers for the Food Insecure in Tampa Bay
DOT National Transportation Integrated Search
2017-09-01
In partnership with the Center for Urban Transportation Research (CUTR) at the University of South Florida (USF), the Transportation Innovation Group informed practical transportation solutions aimed at improved food access in Tampa Bay (Hillsborough...
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Analysis of the Status and Impacts of NCTR Projects
DOT National Transportation Integrated Search
2012-08-01
The National Center for Transit Research (NCTR) at the University of South Florida (USF) assessed the implementation status and identified the outcomes and impacts of the results of 30 Florida Department of Transportation -sponsored NCTR research pro...
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78 FR 29063 - Survey of Urban Rates for Fixed Voice and Fixed Broadband Residential Services
Federal Register 2010, 2011, 2012, 2013, 2014
2013-05-17
... in alternative formats (computer diskette, large print, audio record, and Braille). Persons with... Company Name: Provider FRN (used on MONTH DAY, YEAR Form 477): Provider Study Area Code (if current USF...
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Structure of the human factor VIII C2 domain in complex with the 3E6 inhibitory antibody
Wuerth, Michelle E.; Cragerud, Rebecca K.; Spiegel, P. Clint
2015-11-24
Blood coagulation factor VIII is a glycoprotein cofactor that is essential for the intrinsic pathway of the blood coagulation cascade. Inhibitory antibodies arise either spontaneously or in response to therapeutic infusion of functional factor VIII into hemophilia A patients, many of which are specific to the factor VIII C2 domain. The immune response is largely parsed into “classical” and “non-classical” inhibitory antibodies, which bind to opposing faces cooperatively. In this study, the 2.61 Å resolution structure of the C2 domain in complex with the antigen-binding fragment of the 3E6 classical inhibitory antibody is reported. The binding interface is largely conservedmore » when aligned with the previously determined structure of the C2 domain in complex with two antibodies simultaneously. Further inspection of the B factors for the C2 domain in various X-ray crystal structures indicates that 3E6 antibody binding decreases the thermal motion behavior of surface loops in the C2 domain on the opposing face, thereby suggesting that cooperative antibody binding is a dynamic effect. Furthermore, understanding the structural nature of the immune response to factor VIII following hemophilia A treatment will help lead to the development of better therapeutic reagents.« less
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Diggs, Deacqunita L.; Myers, Jeremy N.; Banks, Leah D.; Niaz, Mohammad S.; Hood, Darryl B.; Roberts, L. Jackson; Ramesh, Aramandla
2013-01-01
In the US alone, around 60,000 lives/year are lost due to colon cancer. Diet and environment have been implicated in the development of sporadic colon tumors. The objective of this study was to determine how dietary fat potentiates the development of colon tumors through altered B(a)P biotransformation, using the Adenomatous polyposis coli with Multiple intestinal neoplasia mouse model. Benzo(a)pyrene was administered to mice through tricaprylin, and unsaturated (USF; peanut oil) and saturated (SF; coconut oil) fats at doses of 50 and 100 μg/kg via oral gavage over a 60-day period. Blood, colon, and liver were collected at the end of exposure period. The expression of B(a)P biotransformation enzymes [cytochrome P450 (CYP)1A1, CYP1B1 and glutathione-S-transferase] in liver and colon were assayed at the level of protein, mRNA and activities. Plasma and tissue samples were analyzed by reverse phase high-performance liquid chromatography for B(a)P metabolites. Additionally, DNA isolated from colon and liver tissues was analyzed for B(a)P-induced DNA adducts by the 32P-postlabeling method using a thin-layer chromatography system. Benzo(a)pyrene exposure through dietary fat altered its metabolic fate in a dose-dependent manner, with 100 μg/kg dose group registering an elevated expression of B(a)P biotransformation enzymes, and greater concentration of B(a)P metabolites, compared to the 50 μg/kg dose group (P<.05). This effect was more pronounced for SF group compared to USF group (P<.05). These findings establish that SF causes sustained induction of B(a)P biotransformation enzymes and extensive metabolism of this toxicant. As a consequence, B(a)P metabolites were generated to a greater extent in colon and liver, whose concentrations also registered a dose-dependent increase. These metabolites were found to bind with DNA and form B(a)P-DNA adducts, which may have contributed to colon tumors in a subchronic exposure regimen. PMID:24231098
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Dash, P K; Tian, L M; Moore, A N
1998-07-07
Axonal injury increases intracellular Ca2+ and cAMP and has been shown to induce gene expression, which is thought to be a key event for regeneration. Increases in intracellular Ca2+ and/or cAMP can alter gene expression via activation of a family of transcription factors that bind to and modulate the expression of CRE (Ca2+/cAMP response element) sequence-containing genes. We have used Aplysia motor neurons to examine the role of CRE-binding proteins in axonal regeneration after injury. We report that axonal injury increases the binding of proteins to a CRE sequence-containing probe. In addition, Western blot analysis revealed that the level of ApCREB2, a CRE sequence-binding repressor, was enhanced as a result of axonal injury. The sequestration of CRE-binding proteins by microinjection of CRE sequence-containing plasmids enhanced axon collateral formation (both number and length) as compared with control plasmid injections. These findings show that Ca2+/cAMP-mediated gene expression via CRE-binding transcription factors participates in the regeneration of motor neuron axons.
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EBP1 is a novel E2F target gene regulated by transforming growth factor-β.
Judah, David; Chang, Wing Y; Dagnino, Lina
2010-11-10
Regulation of gene expression requires transcription factor binding to specific DNA elements, and a large body of work has focused on the identification of such sequences. However, it is becoming increasingly clear that eukaryotic transcription factors can exhibit widespread, nonfunctional binding to genomic DNA sites. Conversely, some of these proteins, such as E2F, can also modulate gene expression by binding to non-consensus elements. E2F comprises a family of transcription factors that play key roles in a wide variety of cellular functions, including survival, differentiation, activation during tissue regeneration, metabolism, and proliferation. E2F factors bind to the Erb3-binding protein 1 (EBP1) promoter in live cells. We now show that E2F binding to the EBP1 promoter occurs through two tandem DNA elements that do not conform to typical consensus E2F motifs. Exogenously expressed E2F1 activates EBP1 reporters lacking one, but not both sites, suggesting a degree of redundancy under certain conditions. E2F1 increases the levels of endogenous EBP1 mRNA in breast carcinoma and other transformed cell lines. In contrast, in non-transformed primary epidermal keratinocytes, E2F, together with the retinoblastoma family of proteins, appears to be involved in decreasing EBP1 mRNA abundance in response to growth inhibition by transforming growth factor-β1. Thus, E2F is likely a central coordinator of multiple responses that culminate in regulation of EBP1 gene expression, and which may vary depending on cell type and context.
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Cooperative DNA binding and sequence discrimination by the Opaque2 bZIP factor.
Yunes, J A; Vettore, A L; da Silva, M J; Leite, A; Arruda, P
1998-01-01
The maize Opaque2 (O2) protein is a basic leucine zipper transcription factor that controls the expression of distinct classes of endosperm genes through the recognition of different cis-acting elements in their promoters. The O2 target region in the promoter of the alpha-coixin gene was analyzed in detail and shown to comprise two closely adjacent binding sites, named O2u and O2d, which are related in sequence to the GCN4 binding site. Quantitative DNase footprint analysis indicated that O2 binding to alpha-coixin target sites is best described by a cooperative model. Transient expression assays showed that the two adjacent sites act synergistically. This synergy is mediated in part by cooperative DNA binding. In tobacco protoplasts, O2 binding at the O2u site is more important for enhancer activity than is binding at the O2d site, suggesting that the architecture of the O2-DNA complex is important for interaction with the transcriptional machinery. PMID:9811800
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Cooperative DNA binding and sequence discrimination by the Opaque2 bZIP factor.
Yunes, J A; Vettore, A L; da Silva, M J; Leite, A; Arruda, P
1998-11-01
The maize Opaque2 (O2) protein is a basic leucine zipper transcription factor that controls the expression of distinct classes of endosperm genes through the recognition of different cis-acting elements in their promoters. The O2 target region in the promoter of the alpha-coixin gene was analyzed in detail and shown to comprise two closely adjacent binding sites, named O2u and O2d, which are related in sequence to the GCN4 binding site. Quantitative DNase footprint analysis indicated that O2 binding to alpha-coixin target sites is best described by a cooperative model. Transient expression assays showed that the two adjacent sites act synergistically. This synergy is mediated in part by cooperative DNA binding. In tobacco protoplasts, O2 binding at the O2u site is more important for enhancer activity than is binding at the O2d site, suggesting that the architecture of the O2-DNA complex is important for interaction with the transcriptional machinery.
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Surtees, Jennifer A; Alani, Eric
2006-07-14
Genetic studies in Saccharomyces cerevisiae predict that the mismatch repair (MMR) factor MSH2-MSH3 binds and stabilizes branched recombination intermediates that form during single strand annealing and gene conversion. To test this model, we constructed a series of DNA substrates that are predicted to form during these recombination events. We show in an electrophoretic mobility shift assay that S. cerevisiae MSH2-MSH3 specifically binds branched DNA substrates containing 3' single-stranded DNA and that ATP stimulates its release from these substrates. Chemical footprinting analyses indicate that MSH2-MSH3 specifically binds at the double-strand/single-strand junction of branched substrates, alters its conformation and opens up the junction. Therefore, MSH2-MSH3 binding to its substrates creates a unique nucleoprotein structure that may signal downstream steps in repair that include interactions with MMR and nucleotide excision repair factors.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Levine, Aaron L; Young, Katherine R
Developers have identified many non-technical barriers to geothermal power development, including access to land. Activities required for accessing land, such as environmental review and private and public leasing can take a considerable amount of time and can delay or prevent project development. This paper discusses the impacts to available geothermal resources and deployment caused by land access challenges, including tribal and cultural resources, environmentally sensitive areas, biological resources, land ownership, federal and state lease queues, and proximity to military installations. In this analysis, we identified challenges that have the potential to prevent development of identified and undiscovered hydrothermal geothermal resources.more » We found that an estimated 400 MW of identified geothermal resource potential and 4,000 MW of undiscovered geothermal resource potential were either unallowed for development or contained one or more significant barriers that could prevent development at the site. Potential improvement scenarios that could be employed to overcome these barriers include (1) providing continuous funding to the U.S. Forest Service (USFS) for processing geothermal leases and permit applications and (2) the creation of advanced environmental mitigation measures. The model results forecast that continuous funding to the USFS could result in deployment of an additional 80 MW of geothermal capacity by 2030 and 124 MW of geothermal capacity by 2050 when compared to the business-as-usual scenario. The creation of advanced environmental mitigation measures coupled with continuous funding to the USFS could result in deployment of an additional 97 MW of geothermal capacity by 2030 and 152 MW of geothermal capacity by 2050 when compared to the business-as-usual scenario. The small impact on potential deployment in these improvement scenarios suggests that these 4,400 MW have other barriers to development in addition to land access. In other words, simply making more resources available for development does not increase deployment; however, impacts to deployment could increase when coupled with other improvements (e.g., permitting, market and/or technology improvements).« less
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An Intercomparison of Large-Extent Tree Canopy Cover Geospatial Datasets
NASA Astrophysics Data System (ADS)
Bender, S.; Liknes, G.; Ruefenacht, B.; Reynolds, J.; Miller, W. P.
2017-12-01
As a member of the Multi-Resolution Land Characteristics Consortium (MRLC), the U.S. Forest Service (USFS) is responsible for producing and maintaining the tree canopy cover (TCC) component of the National Land Cover Database (NLCD). The NLCD-TCC data are available for the conterminous United States (CONUS), coastal Alaska, Hawai'i, Puerto Rico, and the U.S. Virgin Islands. The most recent official version of the NLCD-TCC data is based primarily on reference data from 2010-2011 and is part of the multi-component 2011 version of the NLCD. NLCD data are updated on a five-year cycle. The USFS is currently producing the next official version (2016) of the NLCD-TCC data for the United States, and it will be made publicly-available in early 2018. In this presentation, we describe the model inputs, modeling methods, and tools used to produce the 30-m NLCD-TCC data. Several tree cover datasets at 30-m, as well as datasets at finer resolution, have become available in recent years due to advancements in earth observation data and their availability, computing, and sensors. We compare multiple tree cover datasets that have similar resolution to the NLCD-TCC data. We also aggregate the tree class from fine-resolution land cover datasets to a percent canopy value on a 30-m pixel, in order to compare the fine-resolution datasets to the datasets created directly from 30-m Landsat data. The extent of the tree canopy cover datasets included in the study ranges from global and national to the state level. Preliminary investigation of multiple tree cover datasets over the CONUS indicates a high amount of spatial variability. For example, in a comparison of the NLCD-TCC and the Global Land Cover Facility's Landsat Tree Cover Continuous Fields (2010) data by MRLC mapping zones, the zone-level root mean-square deviation ranges from 2% to 39% (mean=17%, median=15%). The analysis outcomes are expected to inform USFS decisions with regard to the next cycle (2021) of NLCD-TCC production.
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Mazzoni, Esteban O; Mahony, Shaun; Closser, Michael; Morrison, Carolyn A; Nedelec, Stephane; Williams, Damian J; An, Disi; Gifford, David K; Wichterle, Hynek
2013-01-01
Efficient transcriptional programming promises to open new frontiers in regenerative medicine. However, mechanisms by which programming factors transform cell fate are unknown, preventing more rational selection of factors to generate desirable cell types. Three transcription factors, Ngn2, Isl1 and Lhx3, were sufficient to program rapidly and efficiently spinal motor neuron identity when expressed in differentiating mouse embryonic stem cells. Replacement of Lhx3 by Phox2a led to specification of cranial, rather than spinal, motor neurons. Chromatin immunoprecipitation–sequencing analysis of Isl1, Lhx3 and Phox2a binding sites revealed that the two cell fates were programmed by the recruitment of Isl1-Lhx3 and Isl1-Phox2a complexes to distinct genomic locations characterized by a unique grammar of homeodomain binding motifs. Our findings suggest that synergistic interactions among transcription factors determine the specificity of their recruitment to cell type–specific binding sites and illustrate how a single transcription factor can be repurposed to program different cell types. PMID:23872598
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76 FR 14968 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-18
.... 20110076, Draft Supplement, USFS, MT, Grizzly Vegetation and Transportation Management Project, Updated and... Management Actions, Three Rivers Ranger District, Kootenai National Forest, Lincoln County, MT, Comment..., Section 30 Limestone Mining Project, Proposal to Implement Mining Actions, Mystic Ranger District, Black...
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Ecosystem Services Derived from Headwater Catchments
We used data from the USEPA’s wadeable streams assessment (WSA), US Forest Service’s forest inventory and analysis (FIA), and select USFS experimental forests (EF) to investigate potential ecosystems services derived from headwater catchments. C, N, and P inputs to these catchmen...
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A Shared Docking Motif in TRF1 and TRF2 Used for Differential Recruitment of Telomeric Proteins
DOE Office of Scientific and Technical Information (OSTI.GOV)
Chen, Yong; Yang, Yuting; van Overbeek, Megan
2008-05-01
Mammalian telomeres are protected by a six-protein complex: shelterin. Shelterin contains two closely related proteins (TRF1 and TRF2), which recruit various proteins to telomeres. We dissect the interactions of TRF1 and TRF2 with their shared binding partner (TIN2) and other shelterin accessory factors. TRF1 recognizes TIN2 using a conserved molecular surface in its TRF homology (TRFH) domain. However, this same surface does not act as a TIN2 binding site in TRF2, and TIN2 binding to TRF2 is mediated by a region outside the TRFH domain. Instead, the TRFH docking site of TRF2 binds a shelterin accessory factor (Apollo), which doesmore » not interact with the TRFH domain of TRF1. Conversely, the TRFH domain of TRF1, but not of TRF2, interacts with another shelterin-associated factor: PinX1.« less
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Crystal structure of the Rasputin NTF2-like domain from Drosophila melanogaster
DOE Office of Scientific and Technical Information (OSTI.GOV)
Vognsen, Tina, E-mail: tv@farma.ku.dk; Kristensen, Ole, E-mail: ok@farma.ku.dk
2012-03-30
Highlights: Black-Right-Pointing-Pointer The crystal structure of the NTF2-like domain of Rasputin protein is presented. Black-Right-Pointing-Pointer Differences to known ligand binding sites of nuclear transport factor 2 are discussed. Black-Right-Pointing-Pointer A new ligand binding site for the Rasputin and G3BP proteins is proposed. -- Abstract: The crystal structure of the NTF2-like domain of the Drosophila homolog of Ras GTPase SH3 Binding Protein (G3BP), Rasputin, was determined at 2.7 A resolution. The overall structure is highly similar to nuclear transport factor 2: It is a homodimer comprised of a {beta}-sheet and three {alpha}-helices forming a cone-like shape. However, known binding sites formore » RanGDP and FxFG containing peptides show electrostatic and steric differences compared to nuclear transport factor 2. A HEPES molecule bound in the structure suggests a new, and possibly physiologically relevant, ligand binding site.« less
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Nishimura, R; Li, W; Kashishian, A; Mondino, A; Zhou, M; Cooper, J; Schlessinger, J
1993-11-01
Autophosphorylation sites of growth factor receptors with tyrosine kinase activity function as specific binding sites for Src homology 2 (SH2) domains of signaling molecules. This interaction appears to be a crucial step in a mechanism by which receptor tyrosine kinases relay signals to downstream signaling pathways. Nck is a widely expressed protein consisting exclusively of SH2 and SH3 domains, the overexpression of which causes cell transformation. It has been shown that various growth factors stimulate the phosphorylation of Nck and its association with autophosphorylated growth factor receptors. A panel of platelet-derived growth factor (PDGF) receptor mutations at tyrosine residues has been used to identify the Nck binding site. Here we show that mutation at Tyr-751 of the PDGF beta-receptor eliminates Nck binding both in vitro and in living cells. Moreover, the Y751F PDGF receptor mutant failed to mediate PDGF-stimulated phosphorylation of Nck in intact cells. A phosphorylated Tyr-751 is also required for binding of phosphatidylinositol-3 kinase to the PDGF receptor. Hence, the SH2 domains of p85 and Nck share a binding site in the PDGF receptor. Competition experiments with different phosphopeptides derived from the PDGF receptor suggest that binding of Nck and p85 is influenced by different residues around Tyr-751. Thus, a single tyrosine autophosphorylation site is able to link the PDGF receptor to two distinct SH2 domain-containing signaling molecules.
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Klein-Hessling, Stefan; Schneider, Günter; Heinfling, Annette; Chuvpilo, Sergei; Serfling, Edgar
1996-01-01
HMG I(Y) proteins bind to double-stranded A+T oligonucleotides longer than three base pairs. Such motifs form part of numerous NF-AT-binding sites of lymphokine promoters, including the interleukin 4 (IL-4) promoter. NF-AT factors share short homologous peptide sequences in their DNA-binding domain with NF-κB factors and bind to certain NF-κB sites. It has been shown that HMG I(Y) proteins enhance NF-κB binding to the interferon β promoter and virus-mediated interferon β promoter induction. We show that HMG I(Y) proteins exert an opposite effect on the DNA binding of NF-AT factors and the induction of the IL-4 promoter in T lymphocytes. Introduction of mutations into a high-affinity HMG I(Y)-binding site of the IL-4 promoter, which decreased HMG I(Y)-binding to a NF-AT-binding sequence, the Pu-bB (or P) site, distinctly increased the induction of the IL-4 promoter in Jurkat T leukemia cells. High concentrations of HMG I(Y) proteins are able to displace NF-ATp from its binding to the Pu-bB site. High HMG I(Y) concentrations are typical for Jurkat cells and peripheral blood T lymphocytes, whereas El4 T lymphoma cells and certain T helper type 2 cell clones contain relatively low HMG I(Y) concentrations. Our results indicate that HMG I(Y) proteins do not cooperate, but instead compete with NF-AT factors for the binding to DNA even though NF-AT factors share some DNA-binding properties with NF-kB factors. This competition between HMG I(Y) and NF-AT proteins for DNA binding might be due to common contacts with minor groove nucleotides of DNA and may be one mechanism contributing to the selective IL-4 expression in certain T lymphocyte populations, such as T helper type 2 cells. PMID:8986808
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NASA Technical Reports Server (NTRS)
Winchester, S. K.; Selvamurugan, N.; D'Alonzo, R. C.; Partridge, N. C.
2000-01-01
Collagenase-3 mRNA is initially detectable when osteoblasts cease proliferation, increasing during differentiation and mineralization. We showed that this developmental expression is due to an increase in collagenase-3 gene transcription. Mutation of either the activator protein-1 or the runt domain binding site decreased collagenase-3 promoter activity, demonstrating that these sites are responsible for collagenase-3 gene transcription. The activator protein-1 and runt domain binding sites bind members of the activator protein-1 and core-binding factor family of transcription factors, respectively. We identified core-binding factor a1 binding to the runt domain binding site and JunD in addition to a Fos-related antigen binding to the activator protein-1 site. Overexpression of both c-Fos and c-Jun in osteoblasts or core-binding factor a1 increased collagenase-3 promoter activity. Furthermore, overexpression of c-Fos, c-Jun, and core-binding factor a1 synergistically increased collagenase-3 promoter activity. Mutation of either the activator protein-1 or the runt domain binding site resulted in the inability of c-Fos and c-Jun or core-binding factor a1 to increase collagenase-3 promoter activity, suggesting that there is cooperative interaction between the sites and the proteins. Overexpression of Fra-2 and JunD repressed core-binding factor a1-induced collagenase-3 promoter activity. Our results suggest that members of the activator protein-1 and core-binding factor families, binding to the activator protein-1 and runt domain binding sites are responsible for the developmental regulation of collagenase-3 gene expression in osteoblasts.
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Auk Village Recreational Area : Conceptual Parking and Road Improvements Analysis.
DOT National Transportation Integrated Search
2008-07-01
The U.S. Forest Service, Alaska region, Tongass National Forest (USFS), plans to make improvements to the Auk Village Recreation Area (AVRA or recreation area) and specifically Point Louisa Road (the road or roadway) which passes through the AVRA. Th...
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77 FR 4318 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-01-27
..., Clearwater National Forest Travel Planning Project, Proposes to Manage Motorized and Mechanized Travel.../2012, Contact: Heather Berg (208) 476-4541. EIS No. 20120014, Revised Draft EIS, USFS, MT, East Deer Lodge Valley Landscape Restoration Management Project, To Conduct Landscape Restoration Management...
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Effect of test conditions on relative toxicity rankings of fifteen materials
NASA Technical Reports Server (NTRS)
Hilado, C. J.; Cumming, H. J.
1977-01-01
Fifteen materials were evaluated for relative toxicity of pyrolysis effluents, using different test conditions in the USF methodology. Wool fabrics were consistently among the most toxic materials, and polystyrene and polychloroprene flexible foam were consistently among the least toxic materials.
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76 FR 78252 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-16
..., Rubicon Trail Easement and Resource Improvement Project, Construction and Operation, Right-of-Way Grant...: Andrea Catanzaro (409) 766-6346 EIS No. 20110421, Draft EIS, USFS, CA, Greys Mountain Ecological...--La Crosse Transmission System Improvement Project, Proposed Construction and Operation of a 345...
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CCN2/CTGF binds to fibroblast growth factor receptor 2 and modulates its signaling.
Aoyama, Eriko; Kubota, Satoshi; Takigawa, Masaharu
2012-12-14
CCN2 plays a critical role in the development of mesenchymal tissues such as cartilage and bone, and the binding of CCN2 to various cytokines and receptors regulates their signaling.By screening a protein array, we found that CCN2 could bind to fibroblast growth factor receptors (FGFRs) 2 and 3, with a higher affinity toward FGFR2.We ascertained that FGFR2 bound to CCN2 and that the binding of FGFR2 to FGF2 and FGF4 was enhanced by CCN2.CCN2 and FGF2 had a collaborative effect on the phosphorylation of ERK and the differentiation of osteoblastic cells.The present results indicate the biological significance of the binding of CCN2 to FGFR2 in bone metabolism. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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Nag, Ronita; Maity, Manas Kanti; Dasgupta, Maitrayee
2005-11-01
The ABA responsive ABI3 and the auxin responsive ARF family of transcription factors bind the CATGCATG (Sph) and TGTCTC core motifs in ABA and auxin response elements (ABRE and AuxRE), respectively. Several evidences indicate ABI3s to act downstream to auxin too. Because DNA binding domain of ABI3s shows significant overlap with ARFs we enquired whether auxin responsiveness through ABI3s could be mediated by their binding to canonical AuxREs. Investigations were undertaken through in vitro gel mobility shift assays (GMSA) using the DNA binding domain B3 of PvAlf (Phaseolus vulgaris ABI3 like factor) and upstream regions of auxin responsive gene GH3 (-267 to -141) and ABA responsive gene Em (-316 to -146) harboring AuxRE and ABRE, respectively. We demonstrate that B3 domain of PvAlf could bind AuxRE only when B3 was associated with its flanking domain B2 (B2B3). Such strict requirement of B2 domain was not observed with ABRE, where B3 could bind with or without being associated with B2. This dual specificity in DNA binding of ABI3s was also demonstrated with nuclear extracts of cultured cells of Arachis hypogea. Supershift analysis of ABRE and AuxRE bound nuclear proteins with antibodies raised against B2B3 domains of PvAlf revealed that ABI3 associated complexes were detectable in association with both cis elements. Competition GMSA confirmed the same complexes to bind ABRE and AuxRE. This dual specificity of ABI3 like factors in DNA binding targeted to natural promoters responsive to ABA and auxin suggests them to have a potential role in conferring crosstalk between these two phytohormones.
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Measurements of nonlinear Hall-driven reconnection in the reversed field pinch
NASA Astrophysics Data System (ADS)
Tharp, Timothy D.
Complex organisms are able to develop because of the complex regulatory systems that control their gene expression. The first step in this regulation, transcription initiation, is controlled by transcription factors. Transcription factors are modular proteins composed of two distinct domains, the DNA binding domain and the regulatory domain. These molecules are involved in a plethora of important biological processes including embryogenesis, development, cell health, and cancer. Tissue enriched transcription factors Nkx-2.5 and Gata4 are involved in cardiac development and cardiac health. In this thesis the DNA binding specificity of Nkx-2.5 will be analyzed using a high throughput double stranded DNA platform called Cognate Site Identifier (CSI) arrays (Chapter 2). The full DNA binding specificity of Nkx-2.5 and Nkx-2.5 mutants will be visualized using Sequence Specificity Landscapes (SSLs). In Chapter 3, the definition of binding specificity will be investigated by evaluating a number of different DNA binding folds by CSI and SSLs. CSI and SSLs will also be used to evaluate different pyrrole/imidazole hairpin polyamides in order to better characterize these small molecule DNA binding domains. CSI and SSL data will be applied to the genome in order to explain the biological function an artificial transcription factor. Chapter 4 will discuss the mechanism of nonspecific DNA binding. The historical means of predicting DNA binding will be challenged by utilizing high throughput experiments. The effect of salt concentration on both specific and nonspecific binding will also be investigated. Finally, in Chapter 5, a generation of Protein DNA Dimerizer will be discussed. A PDD that regulates transcription on genomic DNA by binding cooperatively with the heart IF Gata4 will be characterized. These studies provide understanding of, and a means to control, how transcription factors sample the endless sea of DNA in the genome in order to regulate gene expression with such wonderful specificity.
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Presnell, Steven R.; Zhang, Lei; Chlebowy, Corrin N.; Al-Attar, Ahmad; Lutz, Charles T.
2012-01-01
KIR2DL4 is unique among human KIR genes in expression, cellular localization, structure, and function, yet the transcription factors required for its expression have not been identified. Using mutagenesis, electrophoretic mobility shift assay, and co-transfection assays, we identified two redundant Runx binding sites in the 2DL4 promoter as essential for constitutive 2DL4 transcription, with contributions by a CRE site and initiator elements. IL-2-and IL-15-stimulated human NK cell lines increased 2DL4 promoter activity, which required functional Runx, CRE, and Ets sites. Chromatin immunoprecipitation experiments show that Runx3 and Ets1 bind the 2DL4 promoter in situ. 2DL4 promoter activity had similar transcription factor requirements in T cells. Runx, CRE, and Ets binding motifs are present in 2DL4 promoters from across primate species, but other postulated transcription factor binding sites are not preserved. Differences between 2DL4 and clonally-restricted KIR promoters suggest a model that explains the unique 2DL4 expression pattern in human NK cells. PMID:22467658
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Some Recent USF Studies at Volcanoes in Central America
NASA Astrophysics Data System (ADS)
McNutt, S. R.
2014-12-01
Scientists at the University of South Florida (USF) have been working in Central America for several decades. Efforts have focused on Physical Volcanology in Nicaragua, GPS in Costa Rica, and assessment of Geothermal projects in El Salvador, amongst others. Two years ago a Seismology Lab was established at USF. Personnel now include three Professors, a Post-Doc, and 4 graduate students. Seismic and GPS networks were installed at Telica Volcano, Nicaragua, in 2010 by Roman, LaFemina and colleagues. Data are recorded on site and recovered several times per year at this persistently restless volcano, which has rates of 5 to 1400 low frequency seismic events per day (Rodgers et al., submitted). Proposals have been submitted to install instruments on other Nicaraguan volcanoes, including seismometers, GPS, infrasound, and lightning sensors. This suite of instruments has proven to be very effective to study a range of volcanic processes. The proposals have not been successful to date (some are pending), and alternative funding sources are being explored. One interesting scientific issue is the presence of strong seasonal effects, specifically a pronounced rainy season and dry season and possible interaction between shallow volcanic processes and surface waters. We are also pursuing a variety of studies that are complementary to the instrumental efforts. One such study is examining volcanic earthquake swarms, with the focus to date on identifying diagnostics. One clear pattern is that peak rates often occur early in swarms, whereas the largest M event occurs late. Additional evidence suggests that the seismic source size grows systematically, especially for events with similar waveforms (families). Recognition of such patterns, linked to processes, may help to improve monitoring and better take advantage of instrumental data to reduce vulnerability from eruptions.
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Kamanga-Sollo, E; Thornton, K J; White, M E; Dayton, W R
2014-10-01
In feedlot steers, estradiol-17β (E2) and combined E2 and trenbolone acetate (a testosterone analog) implants enhance rate and efficiency of muscle growth; and, consequently, these compounds are widely used as growth promoters. Although the positive effects of E2 on rate and efficiency of bovine muscle growth are well established, the mechanisms involved in these effects are not well understood. Combined E2 and trenbolone acetate implants result in significantly increased muscle satellite cell number in feedlot steers. Additionally, E2 treatment stimulates proliferation of cultured bovine satellite cells (BSC). Studies in nonmuscle cells have shown that binding of E2 to G protein-coupled estrogen receptor (GPER)-1 results in activation of matrix metalloproteinases 2 and 9 (MMP2/9) resulting in proteolytic release of heparin binding epidermal growth factor-like growth factor (hbEGF) from the cell surface. Released hbEGF binds to and activates the epidermal growth factor receptor resulting in increased proliferation. To assess if GPER-1, MMP2/9, and/or hbEGF are involved in the mechanism of E2-stimulated BSC proliferation, we have examined the effects of G36 (a specific inhibitor of GPER-1), CRM197 (a specific inhibitor of hbEGF), and MMP-2/MMP-9 Inhibitor II (an inhibitor of MMP2/9 activity) on E2-stimulated BSC proliferation. Inhibition of GPER-1, MMP2/9, or hbEGF suppresses E2-stimulated BSC proliferation (P < 0.001) suggesting that all these are required in order for E2 to stimulate BSC proliferation. These results strongly suggest that E2 may stimulate BSC proliferation by binding to GPER-1 resulting in MMP2/9-catalyzed release of cell membrane-bound hbEGF and subsequent activation of epidermal growth factor receptor by binding of released hbEGF. Copyright © 2014 Elsevier Inc. All rights reserved.
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Factors governing the substitution of La3+ for Ca2+ and Mg2+ in metalloproteins: a DFT/CDM study.
Dudev, Todor; Chang, Li-Ying; Lim, Carmay
2005-03-23
Trivalent lanthanide cations are extensively being used in biochemical experiments to probe various dication-binding sites in proteins; however, the factors governing the binding specificity of lanthanide cations for these binding sites remain unclear. Hence, we have performed systematic studies to evaluate the interactions between La3+ and model Ca2+ - and Mg2+ -binding sites using density functional theory combined with continuum dielectric methods. The calculations reveal the key factors and corresponding physical bases favoring the substitution of trivalent lanthanides for divalent Ca2+ and Mg2+ in holoproteins. Replacing Ca2+ or Mg2+ with La3+ is facilitated by (1) minimizing the solvent exposure and the flexibility of the metal-binding cavity, (2) freeing both carboxylate oxygen atoms of Asp/Glu side chains in the metal-binding site so that they could bind bidentately to La3+, (3) maximizing the number of metal-bound carboxylate groups in buried sites, but minimizing the number of metal-bound carboxylate groups in solvent-exposed sites, and (4) including an Asn/Gln side chain for sites lined with four Asp/Glu side chains. In proteins bound to both Mg2+ and Ca2+, La3+ would prefer to replace Ca2+, as compared to Mg2+. A second Mg2+-binding site with a net positive charge would hamper the Mg2+ --> La3+ exchange, as compared to the respective mononuclear site, although the La3+ substitution of the first native metal is more favorable than the second one. The findings of this work are in accord with available experimental data.
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2009-04-01
Administration ( NASA ) has in place. However, with close coordination among all users, C-band would be available until the primary means to control UAS shifts...Management ( BLM ). White Sands National Monument is located to the southwest. WSMR surrounds the Monument and borders Holloman AFB to the north, west...lands managed by the BLM , U.S. Forest Service (USFS) and State of California. The large population areas of Los Angeles County are located 60 miles
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1994-01-01
Elevation of cAMP can cause gene-specific inhibition of interleukin 2 (IL-2) expression. To investigate the mechanism of this effect, we have combined electrophoretic mobility shift assays and in vivo genomic footprinting to assess both the availability of putative IL-2 transcription factors in forskolin-treated cells and the functional capacity of these factors to engage their sites in vivo. All observed effects of forskolin depended upon protein kinase A, for they were blocked by introduction of a dominant negative mutant subunit of protein kinase A. In the EL4.E1 cell line, we report specific inhibitory effects of cAMP elevation both on NF-kappa B/Rel family factors binding at -200 bp, and on a novel, biochemically distinct "TGGGC" factor binding at -225 bp with respect to the IL-2 transcriptional start site. Neither NF-AT nor AP-1 binding activities are detectably inhibited in gel mobility shift assays. Elevation of cAMP inhibits NF-kappa B activity with delayed kinetics in association with a delayed inhibition of IL-2 RNA accumulation. Activation of cells in the presence of forskolin prevents the maintenance of stable protein- DNA interactions in vivo, not only at the NF-kappa B and TGGGC sites of the IL-2 enhancer, but also at the NF-AT, AP-1, and other sites. This result, and similar results in cyclosporin A-treated cells, imply that individual IL-2 transcription factors cannot stably bind their target sequences in vivo without coengagement of all other distinct factors at neighboring sites. It is proposed that nonhierarchical, cooperative enhancement of binding is a structural basis of combinatorial transcription factor action at the IL-2 locus. PMID:8113685
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77 FR 19281 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-30
..., FL, Central and Southern Florida Project, Broward County Water Preserve Areas, Updates Resulting from Policy Changes that occurred since 2007 Civil Works Board Approval, South Florida Water Management... for this project. EIS No. 20120089, Final EIS, USFS, CA, Greys Mountain Ecological Restoration Project...
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76 FR 76972 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-09
... Supplement, USFS, 00, Programmatic--Kootenai, Idaho Panhandle, and Lolo National Forest Plan Amendments for... has been Identified as the Forest Service's Preferred Alternative, ID, WA, MT, Review Period Ends: 01... Basin Management Unit South Shore Fuel Reduction and Healthy Forest Restoration, To Manage Fuel...
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78 FR 48672 - Environmental Impacts Statements;
Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-09
... Bend Vegetation Management Project, Review Period Ends: 09/19/2013, Contact: Beth Peer 541-383- 4769... Supplement, USFS, ID, Salmon-Challis National Forest Travel Planning and OHV Designation Project, Comment... Mine Project, Comment Period Ends: 09/23/2013, Contact: Shiva Achet 575-234-5924 EIS No. 20130235...
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78 FR 16500 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-15
... and Vegetation Management Project, Comment Period Ends: 04/29/2013, Contact: Marcy Anderson 541-419-0517. EIS No. 20130062, Draft EIS, USFS, NM, Roca Honda Mine Project, Exploration and Mine Development... Seco Riparian Sanctuary Unit Restoration and Pumping Plant/Fish Screen Facility Protection Project...
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76 FR 75543 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-02
... Profile Zones (DFPZs), Lassen National Forest, Almanor Ranger District, Plumas County, CA, Comment Period... Treatment Activities, Updated and New Information, Idaho Panhandle National Forests, Priest Lake Ranger... (937) 257-5899. EIS No. 20110403, Draft EIS, USFS, CA, Creeks II Forest Restoration Project, Proposal...
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White River National Forest Hanging Lake transportation and operations study
DOT National Transportation Integrated Search
2017-05-01
Hanging Lake is a recreation site located on land managed by the U.S. Forest Service (USFS) under the jurisdiction of the White River National Forests Eagle-Holy Cross Ranger District. Due to its increasing popularity over the past few years, the ...
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View of HiattStricklin property privy/outhouse. Note siding and shed design, ...
View of Hiatt-Stricklin property privy/outhouse. Note siding and shed design, facing northwest - Hiatt Property, Privy-Outhouse, West bank of Woof Creek, 400 feet northwest of intersection of U.S.F.S. Roads 651 & 349, Placerville, Boise County, ID
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Cutsforth, G A; Koppaka, V; Krishnaswamy, S; Wu, J R; Mann, K G; Lentz, B R
1996-01-01
The mechanism of binding of blood coagulation cofactor factor Va to acidic-lipid-containing membranes has been addressed. Binding isotherms were generated at room temperature using the change in fluorescence anisotropy of pyrene-labeled bovine factor Va to detect binding to sonicated membrane vesicles containing either bovine brain phosphatidylserine (PS) or 1,2-dioleoyl-3-sn-phosphatidylglycerol (DOPG) in combination with 1-palmitoyl-2-oleoyl-3-sn-phosphatidylcholine (POPC). The composition of the membranes was varied from 0 to 40 mol% for PS/POPC and from 0 to 65 mol % for DOPG/POPC membranes. Fitting the data to a classical Langmuir adsorption model yielded estimates of the dissociation constant (Kd) and the stoichiometry of binding. The values of Kd defined in this way displayed a maximum at low acidic lipid content but were nearly constant at intermediate to high fractions of acidic lipid. Fitting the binding isotherms to a two-process binding model (nonspecific adsorption in addition to binding of acidic lipids to sites on the protein) suggested a significant acidic-lipid-independent binding affinity in addition to occupancy of three protein sites that bind PS in preference to DOPG. Both analyses indicated that interaction of factor Va with an acidic-lipid-containing membrane is much more complex than those of factor Xa or prothrombin. Furthermore, a change in the conformation of bound pyrene-labeled factor Va with surface concentration of acidic lipid was implied by variation of both the saturating fluorescence anisotropy and the binding parameters with the acidic lipid content of the membrane. Finally, the results cannot support the contention that binding occurs through nonspecific adsorption to a patch or domain of acidic lipids in the membrane. Factor Va is suggested to associate with membranes by a complex process that includes both acidic-lipid-specific and acidic-lipid-independent sites and a protein structure change induced by occupancy of acidic-lipid-specific sites on the factor Va molecule. Images FIGURE 5 PMID:8744332
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NASA Technical Reports Server (NTRS)
Spruce, Joseph P.; Ryan, Robert E.; McKellip, Rodney
2008-01-01
The Healthy Forest Restoration Act of 2003 mandated that a national forest threat Early Warning System (EWS) be developed. The USFS (USDA Forest Service) is currently building this EWS. NASA is helping the USFS to integrate remotely sensed data into the EWS, including MODIS data for monitoring forest disturbance at broad regional scales. This RPC experiment assesses the potential of VIIRS (Visible/Infrared Imager/Radiometer Suite) and MODIS (Moderate Resolution Imaging Spectroradiometer) data for contribution to the EWS. In doing so, the RPC project employed multitemporal simulated VIIRS and MODIS data for detecting and monitoring forest defoliation from the non-native Eurasian gypsy moth (Lymantria despar). Gypsy moth is an invasive species threatening eastern U.S. hardwood forests. It is one of eight major forest insect threats listed in the Healthy Forest Restoration Act of 2003. This RPC experiment is relevant to several nationally important mapping applications, including carbon management, ecological forecasting, coastal management, and disaster management
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Butler, William H; Monroe, Ashley; McCaffrey, Sarah
2015-03-01
The Collaborative Forest Landscape Restoration Program (CFLRP), established in 2009, encourages collaborative landscape scale ecosystem restoration efforts on United States Forest Service (USFS) lands. Although the USFS employees have experience engaging in collaborative planning, CFLRP requires collaboration in implementation, a domain where little prior experience can be drawn on for guidance. The purpose of this research is to identify the ways in which CFLRP's collaborative participants and agency personnel conceptualize how stakeholders can contribute to implementation on landscape scale restoration projects, and to build theory on dynamics of collaborative implementation in environmental management. This research uses a grounded theory methodology to explore collaborative implementation from the perspectives and experiences of participants in landscapes selected as part of the CFLRP in 2010. Interviewees characterized collaborative implementation as encompassing three different types of activities: prioritization, enhancing treatments, and multiparty monitoring. The paper describes examples of activities in each of these categories and then identifies ways in which collaborative implementation in the context of CFLRP (1) is both hindered and enabled by overlapping legal mandates about agency collaboration, (2) creates opportunities for expanded accountability through informal and relational means, and, (3) creates feedback loops at multiple temporal and spatial scales through which monitoring information, prioritization, and implementation actions shape restoration work both within and across projects throughout the landscape creating more robust opportunities for adaptive management.
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NASA Astrophysics Data System (ADS)
Butler, William H.; Monroe, Ashley; McCaffrey, Sarah
2015-03-01
The Collaborative Forest Landscape Restoration Program (CFLRP), established in 2009, encourages collaborative landscape scale ecosystem restoration efforts on United States Forest Service (USFS) lands. Although the USFS employees have experience engaging in collaborative planning, CFLRP requires collaboration in implementation, a domain where little prior experience can be drawn on for guidance. The purpose of this research is to identify the ways in which CFLRP's collaborative participants and agency personnel conceptualize how stakeholders can contribute to implementation on landscape scale restoration projects, and to build theory on dynamics of collaborative implementation in environmental management. This research uses a grounded theory methodology to explore collaborative implementation from the perspectives and experiences of participants in landscapes selected as part of the CFLRP in 2010. Interviewees characterized collaborative implementation as encompassing three different types of activities: prioritization, enhancing treatments, and multiparty monitoring. The paper describes examples of activities in each of these categories and then identifies ways in which collaborative implementation in the context of CFLRP (1) is both hindered and enabled by overlapping legal mandates about agency collaboration, (2) creates opportunities for expanded accountability through informal and relational means, and, (3) creates feedback loops at multiple temporal and spatial scales through which monitoring information, prioritization, and implementation actions shape restoration work both within and across projects throughout the landscape creating more robust opportunities for adaptive management.
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Dewi, Vitri; Kwok, Alister; Lee, Stella; Lee, Ming Min; Tan, Yee Mun; Nicholas, Hannah R; Isono, Kyo-ichi; Wienert, Beeke; Mak, Ka Sin; Knights, Alexander J; Quinlan, Kate G R; Cordwell, Stuart J; Funnell, Alister P W; Pearson, Richard C M; Crossley, Merlin
2015-03-27
Krüppel-like factor 3 (KLF3/BKLF), a member of the Krüppel-like factor (KLF) family of transcription factors, is a widely expressed transcriptional repressor with diverse biological roles. Although there is considerable understanding of the molecular mechanisms that allow KLF3 to silence the activity of its target genes, less is known about the signal transduction pathways and post-translational modifications that modulate KLF3 activity in response to physiological stimuli. We observed that KLF3 is modified in a range of different tissues and found that the serine/threonine kinase homeodomain-interacting protein kinase 2 (HIPK2) can both bind and phosphorylate KLF3. Mass spectrometry identified serine 249 as the primary phosphorylation site. Mutation of this site reduces the ability of KLF3 to bind DNA and repress transcription. Furthermore, we also determined that HIPK2 can phosphorylate the KLF3 co-repressor C-terminal binding protein 2 (CtBP2) at serine 428. Finally, we found that phosphorylation of KLF3 and CtBP2 by HIPK2 strengthens the interaction between these two factors and increases transcriptional repression by KLF3. Taken together, our results indicate that HIPK2 potentiates the activity of KLF3. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
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Xu, Emma-Ruoqi; Blythe, Emily E; Fischer, Gerhard; Hyvönen, Marko
2017-07-28
Bone morphogenetic proteins (BMPs) are secreted growth factors that promote differentiation processes in embryogenesis and tissue development. Regulation of BMP signaling involves binding to a variety of extracellular proteins, among which are many von Willebrand factor C (vWC) domain-containing proteins. Although the crystal structure of the complex of crossveinless-2 (CV-2) vWC1 and BMP-2 previously revealed one mode of the vWC/BMP-binding mechanism, other vWC domains may bind to BMP differently. Here, using X-ray crystallography, we present for the first time structures of the vWC domains of two proteins thought to interact with BMP-2: collagen IIA and matricellular protein CCN3. We found that these two vWC domains share a similar N-terminal fold that differs greatly from that in CV-2 vWC, which comprises its BMP-2-binding site. We analyzed the ability of these vWC domains to directly bind to BMP-2 and detected an interaction only between the collagen IIa vWC and BMP-2. Guided by the collagen IIa vWC domain crystal structure and conservation of surface residues among orthologous domains, we mapped the BMP-binding epitope on the subdomain 1 of the vWC domain. This binding site is different from that previously observed in the complex between CV-2 vWC and BMP-2, revealing an alternative mode of interaction between vWC domains and BMPs. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
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View of McKenzieRichey covered well showing log and lumber construction ...
View of McKenzie-Richey covered well showing log and lumber construction and shingles, facing southeast - McKenzie Property, Covered Well, North Bank of Sailor Gulch, 750 feet northwest of intersection of U.S.F.S. Roads 651 & 349, Placerville, Boise County, ID
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Design and evaluation of steel bridges with double composite action
DOT National Transportation Integrated Search
2010-02-01
This report presents findings from a cooperative USF/URS/FDOT research study undertaken to develop design rules for : double composite steel bridges. In the study, a 48 ft long, 16 ft wide, 4 ft. 10 in. deep trapezoidal HPS 70W box section : desig...
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75 FR 21625 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-26
...: Christopher Worthington 775-635-4000. EIS No. 20100136, Final EIS, USFS, 00, Nebraska National Forests and..., Buffalo Gap National Grassland, Oglala National Grassland, Samuel R. McKelvie National Forest, and the Pine Ridge and Bessey Units of the Nebraska National Forest, Fall River, Custer, Pennington, Jackson...
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The potential for alternative transportation at Chimney Rock, San Juan National Forest
DOT National Transportation Integrated Search
2012-05-30
Increased visitation at Chimney Rock in the San Juan National Forest in southwest Colorado has led to increasing interest in the addition of a shuttle system. Piloting a shuttle system at Chimney Rock is a relatively low-cost option that the USFS cou...
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ERIC Educational Resources Information Center
Universal Service Administrative Company, 2009
2009-01-01
The Universal Service Administrative Company (USAC) is an independent, not-for-profit corporation designated as the administrator of the federal Universal Service Fund (USF) by the Federal Communications Commission (FCC). USAC administers the Universal Service Fund and the four Universal Service programs: High Cost, Low Income, Rural Health Care,…
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76 FR 19772 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-08
... Barker 208-735-2072. EIS No. 20110101, Final EIS, USFS, CO, Big Moose Vegetation Management Project... Salvage Project, Proposal to Treat Timer Harvest, Prescribe Fire, and Mechanical Thinning, Uinta-Wasatch... No. 20110107, Final EIS, FHWA, IL, Illinois 336 Corridor Project, (Federal Aid Primary Route 315...
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View of McKenzieRichey barn showing shed design, rolled roofing and ...
View of McKenzie-Richey barn showing shed design, rolled roofing and wood shed roof, facing southwest - McKenzie Property, Barn, North Bank of Sailor Gulch, 750 feet northwest of intersection of U.S.F.S. Roads 651 & 349, Placerville, Boise County, ID
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Code of Federal Regulations, 2010 CFR
2010-10-01
..., usable square footage delivered will be confirmed by either: (1) The Government's measurement of plans... of usable square footage stated in the lease. (c) If the amount of usable square footage delivered is... space delivered and the annual rental will be adjusted as follows: Usable square feet (USF) not...
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Long-term trends in fire behavior and changes in population at risk
Long-term trends in fire behavior and changes in population at risk Rappold AG, Peterson GC, US EPA Matt Jolly, USFS Air pollution regulations and technological advances have successfully reduced emissions of air pollutants from many anthropogenic sources in recent decades. Duri...
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Anderson, Sean M.; Chen, Tom T.; Iruela-Arispe, M. Luisa; Segura, Tatiana
2010-01-01
Growth factors are a class of signaling proteins that direct cell fate through interaction with cell surface receptors. Although a myriad of possible cell fates stem from a growth factor binding to its receptor, the signaling cascades that result in one fate over another are still being elucidated. One possible mechanism by which nature modulates growth factor signaling is through the method of presentation of the growth factor – soluble or immobilized (matrix bound). Here we present the methodology to study signaling of soluble versus immobilized VEGF through VEGFR-2. We have designed a strategy to covalently immobilize VEGF using its heparin-binding domain to orient the molecule (bind) and a secondary functional group to mediate covalent binding (lock). This bind-and-lock approach aims to allow VEGF to assume a bioactive orientation before covalent immobilization. Surface plasmon resonance (SPR) demonstrated heparin and VEGF binding with surface densities of 60 ng/cm2 and 100 pg/cm2, respectively. ELISA experiments confirmed VEGF surface density and showed that electrostatically bound VEGF releases in cell medium and heparin solutions while covalently bound VEGF remains immobilized. Electrostatically bound VEGF and covalently bound VEGF phosphorylate VEGFR-2 in both VEGFR-2 transfected cells and VEGFR-2 endogenously producing cells. HUVECs plated on VEGF functionalized surfaces showed different morphologies between surface-bound VEGF and soluble VEGF. The surfaces synthesized in these studies allow for the study of VEGF/VEGFR-2 signaling induced by covalently bound, electrostatically bound, and soluble VEGF and may provide further insight into the design of materials for the generation of a mature and stable vasculature. PMID:19540581
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PRODORIC2: the bacterial gene regulation database in 2018
Dudek, Christian-Alexander; Hartlich, Juliane; Brötje, David; Jahn, Dieter
2018-01-01
Abstract Bacteria adapt to changes in their environment via differential gene expression mediated by DNA binding transcriptional regulators. The PRODORIC2 database hosts one of the largest collections of DNA binding sites for prokaryotic transcription factors. It is the result of the thoroughly redesigned PRODORIC database. PRODORIC2 is more intuitive and user-friendly. Besides significant technical improvements, the new update offers more than 1000 new transcription factor binding sites and 110 new position weight matrices for genome-wide pattern searches with the Virtual Footprint tool. Moreover, binding sites deduced from high-throughput experiments were included. Data for 6 new bacterial species including bacteria of the Rhodobacteraceae family were added. Finally, a comprehensive collection of sigma- and transcription factor data for the nosocomial pathogen Clostridium difficile is now part of the database. PRODORIC2 is publicly available at http://www.prodoric2.de. PMID:29136200
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Heparin Microparticle Effects on Presentation and Bioactivity of Bone Morphogenetic Protein-2
Hettiaratchi, Marian H.; Miller, Tobias; Temenoff, Johnna S.; Guldberg, Robert E.; McDevitt, Todd C.
2014-01-01
Biomaterials capable of providing localized and sustained presentation of bioactive proteins are critical for effective therapeutic growth factor delivery. However, current biomaterial delivery vehicles commonly suffer from limitations that can result in low retention of growth factors at the site of interest or adversely affect growth factor bioactivity. Heparin, a highly sulfated glycosaminoglycan, is an attractive growth factor delivery vehicle due to its ability to reversibly bind positively charged proteins, provide sustained delivery, and maintain protein bioactivity. This study describes the fabrication and characterization of heparin methacrylamide (HMAm) microparticles for recombinant growth factor delivery. HMAm microparticles were shown to efficiently bind several heparin-binding growth factors (e.g. bone morphogenetic protein-2 (BMP-2), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (FGF-2)), including a wide range of BMP-2 concentrations that exceeds the maximum binding capacity of other common growth factor delivery vehicles, such as gelatin. BMP-2 bioactivity was assessed on the basis of alkaline phosphatase (ALP) activity induced in skeletal myoblasts (C2C12). Microparticles loaded with BMP-2 stimulated comparable C2C12 ALP activity to soluble BMP-2 treatment, indicating that BMP-2-loaded microparticles retain bioactivity and potently elicit a functional cell response. In summary, our results suggest that heparin microparticles stably retain large amounts of bioactive BMP-2 for prolonged periods of time, and that presentation of BMP-2 via heparin microparticles can elicit cell responses comparable to soluble BMP-2 treatment. Consequently, heparin microparticles present an effective method of delivering and spatially retaining growth factors that could be used in a variety of systems to enable directed induction of cell fates and tissue regeneration. PMID:24881028
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Wang, G; Liao, J; Tang, M; Yu, S
2018-02-01
1. Microphthalmia-associated transcription factor (MITF) plays a pivotal role in melanocyte development by regulating the transcription of major pigmentation enzymes (e.g. TYR, TYRP1 and DCT). A single-nucleotide polymorphism (SNP), c.-638T>C, was identified in the MITF promoter, and genotyping of a population (n = 426) revealed that SNP c.-638T>C was associated with skin colour in black-boned chickens. 2. Individuals with genotypes CC and TC exhibited greater MTIF expression than those with genotype TT. Luciferase assays also revealed that genotype CC and TC promoters had higher activity levels than genotype TT. Expression of melanogenesis-related gene (TYR) was higher in the skin of chickens with the CC and CT genotype compared to TT chickens (P < 0.05). 3. Transcription factor-binding site analyses showed that the c.-638C allele contains a putative binding site for transcription factor sterol regulatory element-binding transcription factor 2, aryl hydrocarbon receptor nuclear translocator, transcription factor binding to IGHM enhancer 3 and upstream transcription factor 2. In contrast, the c.-638T allele contains binding sites for Sp3 transcription factor and Krüppel-like factor 1. 4. It was concluded that MITF promoter polymorphisms affected chicken skin colour. SNP c.-638T>C could be used for the marker-assisted selection of skin colour in black-boned chicken breeding.
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Wang, Hao; Jurado, Kellie A; Wu, Xiaolin; Shun, Ming-Chieh; Li, Xiang; Ferris, Andrea L; Smith, Steven J; Patel, Pratiq A; Fuchs, James R; Cherepanov, Peter; Kvaratskhelia, Mamuka; Hughes, Stephen H; Engelman, Alan
2012-12-01
The binding of integrase (IN) to lens epithelium-derived growth factor (LEDGF)/p75 in large part determines the efficiency and specificity of HIV-1 integration. However, a significant residual preference for integration into active genes persists in Psip1 (the gene that encodes for LEDGF/p75) knockout (KO) cells. One other cellular protein, HRP2, harbors both the PWWP and IN-binding domains that are important for LEDGF/p75 co-factor function. To assess the role of HRP2 in HIV-1 integration, cells generated from Hdgfrp2 (the gene that encodes for HRP2) and Psip1/Hdgfrp2 KO mice were infected alongside matched control cells. HRP2 depleted cells supported normal infection, while disruption of Hdgfrp2 in Psip1 KO cells yielded additional defects in the efficiency and specificity of integration. These deficits were largely restored by ectopic expression of either LEDGF/p75 or HRP2. The double-KO cells nevertheless supported residual integration into genes, indicating that IN and/or other host factors contribute to integration specificity in the absence of LEDGF/p75 and HRP2. Psip1 KO significantly increased the potency of an allosteric inhibitor that binds the LEDGF/p75 binding site on IN, a result that was not significantly altered by Hdgfrp2 disruption. These findings help to rule out the host factor-IN interactions as the primary antiviral targets of LEDGF/p75-binding site IN inhibitors.
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77 FR 3265 - Information Collection Being Reviewed by the Federal Communications Commission
Federal Register 2010, 2011, 2012, 2013, 2014
2012-01-23
... business concerns with fewer than 25 employees. The FCC may not conduct or sponsor a collection of... of a currently approved collection. Respondents: Business or other for-profit entities. Number of... previous burden estimates. On November 18, 2011, the Commission adopted the USF/ICC Transformation Order...
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Chelcy Miniat
2013-01-01
The EcoTrends Editorial Committee sorted through vase amounts of historical and ongoing data from 50 ecological sites in the continental United States including Alaska, several islands, and Antarctica to present in a logical format the variables commonly collected. This report presents a subset of data and variables from these sites and illustrates through detailed...
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The "Collaboration in Action: US EPA's Office of Research and Develop - Current Wildfire Research Program" was invited by the USDA's US Forest Service's Scientific Executive Committee to provide USFS scientific leadership active and potential future opportunities for co...
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DOT National Transportation Integrated Search
2007-12-12
At the request of the U.S. Department of Agriculture Forest Service (USFS), the U.S. DOT : Volpe Center conducted a review of the status of Intelligent Transportation Systems (ITS) : planning by the Roaring Fork Transit Authority (RFTA). The assessme...
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Summary of southeastern group breakout sessions
Bob Ford; Charles P. Nicholson
1993-01-01
The breakout sessions held by the southeastern representatives at the Partners In Flight meeting in Colorado were extremely well attended Most states were represented, as well as several federal agencies (including USFS, USFWS, TVA, EPA), and non-government organizations. Two sessions were held, one to discuss a strategy of management by...
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ERIC Educational Resources Information Center
Universal Service Administrative Company, 2009
2009-01-01
This paper presents the activities of the Universal Service Administrative Company (USAC) for 2009. The past year was one of accomplishment for USAC. USAC implemented a host of advances in operations, infrastructure, and outreach in an effort to continue to improve collection and disbursement of the Universal Service Fund (USF) support and to…
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US forest service technical cooperation visit Badia Rangeland and irrigation analysis
USDA-ARS?s Scientific Manuscript database
A US Forest Service (USFS) team comprised of a rangeland management advisor, a dryland water resource, and irrigation specialist, and a Middle East program specialist visited Jordan to provide technical assistance to the Ministry of Agriculture-Water Harvesting Directorate (MoA) and the Hashemite Fu...
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77 FR 60986 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-05
.../2012, Contact: Liana Liu 360-753-9553. EIS No. 20120312, Final EIS, USFS, NV, Geothermal Leasing on the Humboldt-Toiyabe National Forest, To Facilitate the Development and Production of Geothermal Energy, Ely..., Contact: Jerald Weaver 360-378-2223. EIS No. 20120314, Draft EIS, WAPA, WY, Hermosa West Wind Energy...
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Deborah J. Chavez; Patricia C. Winter; James D. Absher
2008-01-01
In 1987, the Pacific Southwest Research Station (PSW) of the U.S. Department of Agriculture, Forest Service (USFS), chartered a research work unit to examine outdoor recreation in the wildland-urban interface. The need for the work unit was identified by the four forest supervisors in southern California, from the Angeles National Forest, the Cleveland National Forest...
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77 FR 9652 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-17
... Road-Related Impacts to Wildlife, Fish, Soil, and Water Resources and Restoration of 2010 Forest Plan..., Amendment 18A to the Fishery Management Plan for the Snapper-Grouper Fishery of the South Atlantic Region...-9235. EIS No. 20120034, Draft EIS, USFS, CA, Harris Vegetation Management Project, To Improve Forest...
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McGlade, C J; Ellis, C; Reedijk, M; Anderson, D; Mbamalu, G; Reith, A D; Panayotou, G; End, P; Bernstein, A; Kazlauskas, A
1992-01-01
The binding of cytoplasmic signaling proteins such as phospholipase C-gamma 1 and Ras GTPase-activating protein to autophosphorylated growth factor receptors is directed by their noncatalytic Src homology region 2 (SH2) domains. The p85 alpha regulatory subunit of phosphatidylinositol (PI) 3-kinase, which associates with several receptor protein-tyrosine kinases, also contains two SH2 domains. Both p85 alpha SH2 domains, when expressed individually as fusion proteins in bacteria, bound stably to the activated beta receptor for platelet-derived growth factor (PDGF). Complex formation required PDGF stimulation and was dependent on receptor tyrosine kinase activity. The bacterial p85 alpha SH2 domains recognized activated beta PDGF receptor which had been immobilized on a filter, indicating that SH2 domains contact autophosphorylated receptors directly. Several receptor tyrosine kinases within the PDGF receptor subfamily, including the colony-stimulating factor 1 receptor and the Steel factor receptor (Kit), also associate with PI 3-kinase in vivo. Bacterially expressed SH2 domains derived from the p85 alpha subunit of PI 3-kinase bound in vitro to the activated colony-stimulating factor 1 receptor and to Kit. We infer that the SH2 domains of p85 alpha bind to high-affinity sites on these receptors, whose creation is dependent on receptor autophosphorylation. The SH2 domains of p85 are therefore primarily responsible for the binding of PI 3-kinase to activated growth factor receptors. Images PMID:1372092
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Chang, Chun-Chun; Hsu, Hao-Jen; Yen, Jui-Hung; Lo, Shih-Yen
2017-01-01
Hepatitis C virus (HCV) is a species-specific pathogenic virus that infects only humans and chimpanzees. Previous studies have indicated that interactions between the HCV E2 protein and CD81 on host cells are required for HCV infection. To determine the crucial factors for species-specific interactions at the molecular level, this study employed in silico molecular docking involving molecular dynamic simulations of the binding of HCV E2 onto human and rat CD81s. In vitro experiments including surface plasmon resonance measurements and cellular binding assays were applied for simple validations of the in silico results. The in silico studies identified two binding regions on the HCV E2 loop domain, namely E2-site1 and E2-site2, as being crucial for the interactions with CD81s, with the E2-site2 as the determinant factor for human-specific binding. Free energy calculations indicated that the E2/CD81 binding process might follow a two-step model involving (i) the electrostatic interaction-driven initial binding of human-specific E2-site2, followed by (ii) changes in the E2 orientation to facilitate the hydrophobic and van der Waals interaction-driven binding of E2-site1. The sequence of the human-specific, stronger-binding E2-site2 could serve as a candidate template for the future development of HCV-inhibiting peptide drugs. PMID:28481946
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Chen, Zeming; Kolokoltsov, Andrey A.; Wang, Jia; Adhikary, Shramika; Lorinczi, Marta; Elferink, Lisa A.
2012-01-01
For retroviruses such as HIV-1 and murine leukemia virus (MLV), active receptor recruitment and trafficking occur during viral entry. However, the underlying mechanisms and cellular factors involved in the process are largely uncharacterized. The viral receptor for ecotropic MLV (eMLV), a classical model for retrovirus infection mechanisms and pathogenesis, is mouse cationic amino acid transporter 1 (mCAT-1). Growth factor receptor-bound protein 2 (GRB2) is an adaptor protein that has been shown to couple cell surface receptors, such as epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor, to intracellular signaling events. Here we examined if GRB2 could also play a role in controlling infection by retroviruses by affecting receptor function. The GRB2 RNA interference (RNAi)-mediated suppression of endogenous GRB2 resulted in a consistent and significant reduction of virus binding and membrane fusion. The binding between eMLV and cells promoted increased GRB2–mCAT-1 interactions, as detected by immunoprecipitation. Consistently, the increased colocalization of GRB2 and mCAT-1 signals was detected by confocal microscopy. This association was time dependent and paralleled the kinetics of cell-virus membrane fusion. Interestingly, unlike the canonical binding pattern seen for GRB2 and growth factor receptors, GRB2–mCAT-1 binding does not depend on the GRB2-SH2 domain-mediated recognition of tyrosine phosphorylation on the receptor. The inhibition of endogenous GRB2 led to a reduction in surface levels of mCAT-1, which was detected by immunoprecipitation and by a direct binding assay using a recombinant MLV envelope protein receptor binding domain (RBD). Consistent with this observation, the expression of a dominant negative GRB2 mutant (R86K) resulted in the sequestration of mCAT-1 from the cell surface into intracellular vesicles. Taken together, these findings suggest a novel role for GRB2 in ecotropic MLV entry and infection by facilitating mCAT-1 trafficking. PMID:22090132
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Chen, Zeming; Kolokoltsov, Andrey A; Wang, Jia; Adhikary, Shramika; Lorinczi, Marta; Elferink, Lisa A; Davey, Robert A
2012-02-01
For retroviruses such as HIV-1 and murine leukemia virus (MLV), active receptor recruitment and trafficking occur during viral entry. However, the underlying mechanisms and cellular factors involved in the process are largely uncharacterized. The viral receptor for ecotropic MLV (eMLV), a classical model for retrovirus infection mechanisms and pathogenesis, is mouse cationic amino acid transporter 1 (mCAT-1). Growth factor receptor-bound protein 2 (GRB2) is an adaptor protein that has been shown to couple cell surface receptors, such as epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor, to intracellular signaling events. Here we examined if GRB2 could also play a role in controlling infection by retroviruses by affecting receptor function. The GRB2 RNA interference (RNAi)-mediated suppression of endogenous GRB2 resulted in a consistent and significant reduction of virus binding and membrane fusion. The binding between eMLV and cells promoted increased GRB2-mCAT-1 interactions, as detected by immunoprecipitation. Consistently, the increased colocalization of GRB2 and mCAT-1 signals was detected by confocal microscopy. This association was time dependent and paralleled the kinetics of cell-virus membrane fusion. Interestingly, unlike the canonical binding pattern seen for GRB2 and growth factor receptors, GRB2-mCAT-1 binding does not depend on the GRB2-SH2 domain-mediated recognition of tyrosine phosphorylation on the receptor. The inhibition of endogenous GRB2 led to a reduction in surface levels of mCAT-1, which was detected by immunoprecipitation and by a direct binding assay using a recombinant MLV envelope protein receptor binding domain (RBD). Consistent with this observation, the expression of a dominant negative GRB2 mutant (R86K) resulted in the sequestration of mCAT-1 from the cell surface into intracellular vesicles. Taken together, these findings suggest a novel role for GRB2 in ecotropic MLV entry and infection by facilitating mCAT-1 trafficking.
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Vadivel, Kanagasabai; Ponnuraj, Sathya-Moorthy; Kumar, Yogesh; Zaiss, Anne K.; Bunce, Matthew W.; Camire, Rodney M.; Wu, Ling; Evseenko, Denis; Herschman, Harvey R.; Bajaj, Madhu S.; Bajaj, S. Paul
2014-01-01
Tissue factor pathway inhibitor-2 (TFPI-2) is a homologue of TFPI-1 and contains three Kunitz-type domains and a basic C terminus region. The N-terminal domain of TFPI-2 is the only inhibitory domain, and it inhibits plasma kallikrein, factor XIa, and plasmin. However, plasma TFPI-2 levels are negligible (≤20 pm) in the context of influencing clotting or fibrinolysis. Here, we report that platelets contain significant amounts of TFPI-2 derived from megakaryocytes. We employed RT-PCR, Western blotting, immunohistochemistry, and confocal microscopy to determine that platelets, MEG-01 megakaryoblastic cells, and bone marrow megakaryocytes contain TFPI-2. ELISA data reveal that TFPI-2 binds factor V (FV) and partially B-domain-deleted FV (FV-1033) with Kd ∼9 nm and binds FVa with Kd ∼100 nm. Steady state analysis of surface plasmon resonance data reveal that TFPI-2 and TFPI-1 bind FV-1033 with Kd ∼36–48 nm and bind FVa with Kd ∼252–456 nm. Further, TFPI-1 (but not TFPI-1161) competes with TFPI-2 in binding to FV. These data indicate that the C-terminal basic region of TFPI-2 is similar to that of TFPI-1 and plays a role in binding to the FV B-domain acidic region. Using pull-down assays and Western blots, we show that TFPI-2 is associated with platelet FV/FVa. TFPI-2 (∼7 nm) in plasma of women at the onset of labor is also, in part, associated with FV. Importantly, TFPI-2 in platelets and in plasma of pregnant women inhibits FXIa and tissue-type plasminogen activator-induced clot fibrinolysis. In conclusion, TFPI-2 in platelets from normal or pregnant subjects and in plasma from pregnant women binds FV/Va and regulates intrinsic coagulation and fibrinolysis. PMID:25262870
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COUP-TF1 antagonizes Nkx2.5-mediated activation of the calreticulin gene during cardiac development.
Guo, L; Lynch, J; Nakamura, K; Fliegel, L; Kasahara, H; Izumo, S; Komuro, I; Agellon, L B; Michalak, M
2001-01-26
Calreticulin, a Ca(2+) binding chaperone of the endoplasmic reticulum, is also highly expressed in the embryonic heart, and knockout of the calreticulin gene is lethal during embryogenesis because of impaired cardiac development. The protein is down-regulated after birth, and elevated expression of calreticulin in newborn hearts is associated with severe cardiac pathology and death. Here we show that the transcription factor Nkx2.5 activates expression of the calreticulin gene in the heart. Binding of chicken ovalbumin upstream promoter-transcription factor 1 to the Nkx2.5 binding site suppresses transcription from the calreticulin promoter. Nkx2.5 and chicken ovalbumin upstream promoter-transcription factor 1 play antagonistic roles in regulating the expression of calreticulin during cardiac development. These studies indicate that cardiac-specific transcription factor Nkx2.5 plays a central role in activating calreticulin expression and that there is a cooperation between chicken ovalbumin upstream promoter-transcription factor 1 and Nkx2.5 at the calreticulin promoter.
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Noda, Chieko; Narita, Yohei; Watanabe, Takahiro; Yoshida, Masahiro; Ashio, Keiji; Sato, Yoshitaka; Goshima, Fumi; Kanda, Teru; Yoshiyama, Hironori; Tsurumi, Tatsuya; Kimura, Hiroshi
2016-01-01
ABSTRACT Latent membrane protein 1 (LMP1) is a major oncogene essential for primary B cell transformation by Epstein-Barr virus (EBV). Previous studies suggested that some transcription factors, such as PU.1, RBP-Jκ, NF-κB, and STAT, are involved in this expression, but the underlying mechanism is unclear. Here, we identified binding sites for PAX5, AP-2, and EBF in the proximal LMP1 promoter (ED-L1p). We first confirmed the significance of PU.1 and POU domain transcription factor binding for activation of the promoter in latency III. We then focused on the transcription factors AP-2 and early B cell factor (EBF). Interestingly, among the three AP-2-binding sites in the LMP1 promoter, two motifs were also bound by EBF. Overexpression, knockdown, and mutagenesis in the context of the viral genome indicated that AP-2 plays an important role in LMP1 expression in latency II in epithelial cells. In latency III B cells, on the other hand, the B cell-specific transcription factor EBF binds to the ED-L1p and activates LMP1 transcription from the promoter. IMPORTANCE Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) is crucial for B cell transformation and oncogenesis of other EBV-related malignancies, such as nasopharyngeal carcinoma and T/NK lymphoma. Its expression is largely dependent on the cell type or condition, and some transcription factors have been implicated in its regulation. However, these previous reports evaluated the significance of specific factors mostly by reporter assay. In this study, we prepared point-mutated EBV at the binding sites of such transcription factors and confirmed the importance of AP-2, EBF, PU.1, and POU domain factors. Our results will provide insight into the transcriptional regulation of the major oncogene LMP1. PMID:26819314
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Wang, Jun; Lee, Seungsoo; Teh, Charis En-Yi; Bunting, Karen; Ma, Lina; Shannon, M Frances
2009-03-01
Activation of T cells leads to the induction of many cytokine genes that are required for appropriate immune responses, including IL-2, a key cytokine for T cell proliferation and homeostasis. The activating transcription factors such as nuclear factor of activated T cells, nuclear factor kappaB/Rel and activated protein-1 family members that regulate inducible IL-2 gene expression have been well documented. However, negative regulation of the IL-2 gene is less studied. Here we examine the role of zinc finger E-box-binding protein (ZEB) 1, a homeodomain/Zn finger transcription factor, as a repressor of IL-2 gene transcription. We show here that ZEB1 is expressed in non-stimulated and stimulated T cells and using chromatin immunoprecipitation assays we show that ZEB1 binds to the IL-2 promoter. Over-expression of ZEB1 can repress IL-2 promoter activity, as well as endogenous IL-2 mRNA production in EL-4 T cells, and this repression is dependent on the ZEB-binding site at -100. ZEB1 cooperates with the co-repressor C-terminal-binding protein (CtBP) 2 and with histone deacetylase 1 to repress the IL-2 promoter and this cooperation depends on the ZEB-binding site in the promoter as well as the Pro-X-Asp-Leu-Ser protein-protein interaction domain in CtBP2. Thus, ZEB1 may function to recruit a repressor complex to the IL-2 promoter.
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Barcelona, Pablo F.
2015-01-01
Nerve growth factor (NGF) is generated from a precursor, proNGF, that is proteolytically processed. NGF preferentially binds a trophic tyrosine kinase receptor, TrkA, while proNGF binds a neurotrophin receptor (NTR), p75NTR, that can have neurotoxic activity. Previously, we along with others showed that the soluble protein α2-macroglobulin (α2M) is neurotoxic. Toxicity is due in part to α2M binding to NGF and inhibiting trophic activity, presumably by preventing NGF binding to TrkA. However, the mechanisms remained unclear. Here, we show ex vivo and in vivo three mechanisms for α2M neurotoxicity. First, unexpectedly the α2M-NGF complexes do bind TrkA receptors but do not induce TrkA dimerization or activation, resulting in deficient trophic support. Second, α2M makes stable complexes with proNGF, conveying resistance to proteolysis that results in more proNGF and less NGF. Third, α2M-proNGF complexes bind p75NTR and are more potent agonists than free proNGF, inducing tumor necrosis factor alpha (TNF-α) production. Hence, α2M regulates proNGF/p75NTR positively and mature NGF/TrkA negatively, causing neuronal death ex vivo. These three mechanisms are operative in vivo, and α2M causes neurodegeneration in a p75NTR- and proNGF-dependent manner. α2M could be exploited as a therapeutic target, or as a modifier of neurotrophin signals. PMID:26217017
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DOE Office of Scientific and Technical Information (OSTI.GOV)
F. Robert Tabita
The ocean/atmosphere interface is the major conduit for the entry of atmospheric CO2 into oceanic carbon pools that can lead to sequestration or recycled release. The surface layers of the temperate and tropical oceans are often too oligotrophic to result in significant primary production that might lead to carbon sequestration. However, nutrient-rich river plumes can alter the primary production schemes of oligotrophic ocean basins, resulting in increased phytoplankton biomass and carbon fixation. The ultimate goal of this proposal is to understand these carbon cycling processes in major river plumes from the molecular processes involved in biological DIC uptake to contributionmore » to basin-wide production and potential sequestration. Our research efforts include a field component to answer the questions raised concerning DIC in plumes entering ocean basins and an intensive genomics approach to understanding these processes on the cellular level using genomic fragments obtained from plume biota. This project is actually composed of 3 separate PI-initiated projects, including projects at the University of South Florida (USF) College of Marine Science, the University of Puerto Rico, and The Ohio State University. This report concerns research conducted at The Ohio State University and studies performed in collaboration with USF. In order to understand what might occur in the field, two model sysytems were studied in the laboratory. Carbon fixation in the unicellular cyanobacterium Synechococcus sp Strain PCC 7002 took place mainly through the CBB pathway. Nitrogen nutrition in cyanobacteria is regulated by NtcA, a transcriptional regulatory protein. We show that the rubisco activity and gene (rbcL) expression were not affected when cells were exposed to prolonged periods of nitrogen stress, however cells appear to use intracellular nitrogen reserves during nitrogen starvation. Transcripts of the global transcriptional regulator NtcA are expressed under nitrogen starved and nitrogen replete (nitrate or ammonia) growth conditions, with slight decrease in transcription in the presence of ammonia. These results suggest that intracellular levels of NtcA do not directly affect carbon metabolism. Gene expression of the other nitrogen regulatory signal transducer, encoded by glnB was also studied. The glnB gene was highly transcribed in nitrogen-limited cells compared to nitrogen depleted growth conditions. Therefore in the cyanobacterium Synechococcus sp PCC 7002, nitrogen does not affect the metabolic potential and carbon fixation. The NtcA regulator behaved differently and studies indicate that the product of the ntcA gene (NtcA) has an indirect effect on ca rbon assimilation and the genes involved in the carbon concentrating mechanism of strain 7002. The product of the ccmM gene plays an important role in carboxysome assembly and inorganic carbon transport within the cell. We hypothesized that under nitrogen limiting conditions the transcriptional regulator NtcA binds at the region upstream of ccmM, near the transcription start site, and blocks the transcription of ccmM. This hypothesis was experimentally proven. In another study, with USF researchers, we performed experiments in situ on RubisCO espression. To determine the relationship between expression of the major gene in carbon fixation, we evaluated rbcL mRNA abundance using novel quantitative PCR assays, phytoplankton cell analyses, photophysiological parameters, and pCO2 in and around the Mississippi River plume (MRP) in the Gulf of Mexico. Lower salinity (30–32) stations were dominated by rbcL mRNA concentrations from heterokonts; i.e., diatoms and pelagophytes, which were at least an order of magnitude greater than haptophytes, a-Synechococcus or high-light Prochlorococcus. However, rbcL transcript abundances were similar among these groups at oligotrophic stations (salinity 34–36). Diatom cell counts and heterokont rbcL RNA showed a strong negative correlation to seawater pCO2. While Prochlorococcus cells did not exhibit a large difference between low and high pCO2 water, Prochlorococcus rbcL RNA concentrations had a strong positive correlation to pCO2, suggesting a very low level of RuBisCO RNA transcription among Prochlorococcus in the plume waters, possibly due to their relatively poor carbon concentrating mechanisms (CCMs). These results provide molecular evidence that diatom/pelagophyte productivity is largely responsible for the large CO2 drawdown occurring in the MRP, based on the cooccurrence of elevated RuBisCO gene transcript concentrations from this group and reduced seawater pCO2 levels. This may partly be due to efficient CCMs that enable heterokont eukaryotes such as diatoms to continue fixing CO2 in the face of strong CO2 drawdown. This work represents the first attempt to relate in situ microbial gene expression to contemporaneous CO2 flux measurements in the ocean.« less
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Automated system for smoke dispersion prediction due to wild fires in Alaska
NASA Astrophysics Data System (ADS)
Kulchitsky, A.; Stuefer, M.; Higbie, L.; Newby, G.
2007-12-01
Community climate models have enabled development of specific environmental forecast systems. The University of Alaska (UAF) smoke group was created to adapt a smoke forecast system to the Alaska region. The US Forest Service (USFS) Missoula Fire Science Lab had developed a smoke forecast system based on the Weather Research and Forecasting (WRF) Model including chemistry (WRF/Chem). Following the successful experience of USFS, which runs their model operationally for the contiguous U.S., we develop a similar system for Alaska in collaboration with scientists from the USFS Missoula Fire Science Lab. Wildfires are a significant source of air pollution in Alaska because the climate and vegetation favor annual summer fires that burn huge areas. Extreme cases occurred in 2004, when an area larger than Maryland (more than 25000~km2) burned. Small smoke particles with a diameter less than 10~μm can penetrate deep into lungs causing health problems. Smoke also creates a severe restriction to air transport and has tremendous economical effect. The smoke dispersion and forecast system for Alaska was developed at the Geophysical Institute (GI) and the Arctic Region Supercomputing Center (ARSC), both at University of Alaska Fairbanks (UAF). They will help the public and plan activities a few days in advance to avoid dangerous smoke exposure. The availability of modern high performance supercomputers at ARSC allows us to create and run high-resolution, WRF-based smoke dispersion forecast for the entire State of Alaska. The core of the system is a Python program that manages the independent pieces. Our adapted Alaska system performs the following steps \\begin{itemize} Calculate the medium-resolution weather forecast using WRF/Met. Adapt the near real-time satellite-derived wildfire location and extent data that are received via direct broadcast from UAF's "Geographic Information Network of Alaska" (GINA) Calculate fuel moisture using WRF forecasts and National Fire Danger Rating System (NFDRS) fuel maps Calculate smoke emission components using a first order fire emission model Model the smoke plume rise yielding a vertically distribution that accounts for one-dimensional (vertical) concentrations of smoke constituents in the atmosphere above the fire Run WRF/Chem at high resolution for the forecast Use standard graphical tools to provide accessible smoke dispersion The system run twice each day at ARSC. The results will be freely available from a dedicated wildfire smoke web portal at ARSC.
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Kunsan AB, Korea. Revised Uniform Summary of Surface Weather Observations (RUSSWO). Parts A-F
1981-05-01
A.. . .... .. . . . . . ... .. . ... _ GLOBAL CLI AT L-O Y BRA CH usF ,cCEILING VERSUS VISIBILITY 4 3 2 1 9 K U N S A N A K O 6 8 -7 0 97 3...77,.C 77.S 78,3 78,3 78*4 7805 78,5 7845 78,5 79,0 79,3S0 oo 70 74o 7417 791( 80.3 81. 8W. 81.6 NOV 81.9 81.9 81*9 1.9 82*3 82.6 > 35oo 74.5 79*1 796
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Human blood-brain barrier insulin-like growth factor receptor
DOE Office of Scientific and Technical Information (OSTI.GOV)
Duffy, K.R.; Pardridge, W.M.; Rosenfeld, R.G.
1988-02-01
Insulin-like growth factor (IGF)-1 and IGF-2, may be important regulatory molecules in the CNS. Possible origins of IGFs in brain include either de novo synthesis or transport of circulating IGFs from blood into brain via receptor mediated transcytosis mechanisms at the brain capillary endothelial wall, ie, the blood-brain barrier (BBB). In the present studies, isolated human brain capillaries are used as an in vitro model system of the human BBB and the characteristics of IGF-1 or IGF-2 binding to this preparation were assessed. The total binding of IGF-2 at 37 degrees C exceeded 130% per mg protein and was threefoldmore » greater than the total binding for IGF-1. However, at 37 degrees C nonsaturable binding equaled total binding, suggesting that endocytosis is rate limiting at physiologic temperatures. Binding studies performed at 4 degrees C slowed endocytosis to a greater extent than membrane binding, and specific binding of either IGF-1 or IGF-2 was detectable. Scatchard plots for either peptide were linear and the molar dissociation constant of IGF-1 and IGF-2 binding was 2.1 +/- 0.4 and 1.1 +/- 0.1 nmol/L, respectively. Superphysiologic concentrations of porcine insulin inhibited the binding of both IGF-1 (ED50 = 2 micrograms/mL) and IGF-2 (ED50 = 0.5 microgram/mL). Affinity cross linking of /sup 125/I-IGF-1, /sup 125/I-IGF-2, and /sup 125/I-insulin to isolated human brain capillaries was performed using disuccinimidylsuberate (DSS). These studies revealed a 141 kd binding site for both IGF-1 and IGF-2, and a 133 kd binding site for insulin.« less
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Kucharski, Amir N; Scott, Caitlin E; Davis, Jonathan P; Kekenes-Huskey, Peter M
2016-08-25
Parvalbumin (PV) is a globular calcium (Ca(2+))-selective protein expressed in a variety of biological tissues. Our computational studies of the rat β-parvalbumin (β-PV) isoform seek to elucidate the molecular thermodynamics of Ca(2+) versus magnesium (Mg(2+)) binding at the protein's two EF-hand motifs. Specifically, we have utilized molecular dynamics (MD) simulations and a mean-field electrolyte model (mean spherical approximation (MSA) theory) to delineate how the EF-hand scaffold controls the "local" thermodynamics of Ca(2+) binding selectivity over Mg(2+). Our MD simulations provide the probability density of metal-chelating oxygens within the EF-hand scaffolds for both Ca(2+) and Mg(2+), as well the conformational strain induced by Mg(2+) relative to Ca(2+) binding. MSA theory utilizes the binding domain oxygen and charge distributions to predict the chemical potential of ion binding, as well as their corresponding concentrations within the binding domain. We find that the electrostatic and steric contributions toward ion binding were similar for Mg(2+) and Ca(2+), yet the latter was 5.5 kcal/mol lower in enthalpy when internal strain within the EF hand was considered. We therefore speculate that beyond differences in dehydration energies for the Ca(2+) versus Mg(2+), strain induced in the β-PV EF hand by cation binding significantly contributes to the nearly 10,000-fold difference in binding affinity reported in the literature. We further complemented our analyses of local factors governing cation binding selectivity with whole-protein (global) contributions, such as interhelical residue-residue contacts and solvent exposure of hydrophobic surface. These contributions were found to be comparable for both Ca(2+)- and Mg(2+)-bound β-PV, which may implicate local factors, EF-hand strain, and dehydration, in providing the primary means of selectivity. We anticipate these methods could be used to estimate metal binding thermodynamics across a broad range of PV sequence homologues and EF-hand-containing, Ca(2+) binding proteins.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Barber, Claire; Netherton, Chris; Goatley, Lynnett
The African swine fever virus DP71L protein recruits protein phosphatase 1 (PP1) to dephosphorylate the translation initiation factor 2α (eIF2α) and avoid shut-off of global protein synthesis and downstream activation of the pro-apoptotic factor CHOP. Residues V16 and F18A were critical for binding of DP71L to PP1. Mutation of this PP1 binding motif or deletion of residues between 52 and 66 reduced the ability of DP71L to cause dephosphorylation of eIF2α and inhibit CHOP induction. The residues LSAVL, between 57 and 61, were also required. PP1 was co-precipitated with wild type DP71L and the mutant lacking residues 52- 66 ormore » the LSAVL motif, but not with the PP1 binding motif mutant. The residues in the LSAVL motif play a critical role in DP71L function but do not interfere with binding to PP1. Instead we propose these residues are important for DP71L binding to eIF2α. - Highlights: •The African swine fever virus DP71L protein recruits protein phosphatase 1 (PP1) to dephosphorylate translation initiation factor eIF2α (eIF2α). •The residues V{sup 16}, F{sup 18} of DP71L are required for binding to the α, β and γ isoforms of PP1 and for DP71L function. •The sequence LSAVL downstream from the PP1 binding site (residues 57–61) are also important for DP71L function. •DP71L mutants of the LSAVL sequence retain ability to co-precipitate with PP1 showing these sequences have a different role to PP1 binding.« less
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Zheng, Zhaoqing; Ambigapathy, Ganesh; Keifer, Joyce
2017-01-01
MECP2 mutations underlying Rett syndrome cause widespread misregulation of gene expression. Functions for MeCP2 other than transcriptional are not well understood. In an ex vivo brain preparation from the pond turtle Trachemys scripta elegans, an intraexonic splicing event in the brain-derived neurotrophic factor (BDNF) gene generates a truncated mRNA transcript in naïve brain that is suppressed upon classical conditioning. MeCP2 and its partners, splicing factor Y-box binding protein 1 (YB-1) and methylcytosine dioxygenase 1 (Tet1), bind to BDNF chromatin in naïve but dissociate during conditioning; the dissociation correlating with decreased DNA methylation. Surprisingly, conditioning results in new occupancy of BDNF chromatin by DNA insulator protein CCCTC-binding factor (CTCF), which is associated with suppression of splicing in conditioning. Knockdown of MeCP2 shows it is instrumental for splicing and inhibits Tet1 and CTCF binding thereby negatively impacting DNA methylation and conditioning-dependent splicing regulation. Thus, mutations in MECP2 can have secondary effects on DNA methylation and alternative splicing. DOI: http://dx.doi.org/10.7554/eLife.25384.001 PMID:28594324
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Nordqvist, A C; Peyrard, M; Pettersson, H; Mathiesen, T; Collins, V P; Dumanski, J P; Schalling, M
1997-07-01
Insulin-like growth factors (IGFs) I and II have been implicated as autocrine or paracrine growth promoters. These growth factors bind to specific receptors, and the response is modulated by interaction with IGF-binding proteins (IGFBPs). We observed a strong correlation between anaplastic/atypical histopathology and a high IGF-II/IGFBP-2 mRNA ratio in a set of 68 sporadic meningiomas. A strong correlation was also found between clinical outcome and IGF-II/IGFBP-2 ratio, whereas previously used histochemical markers were less correlated to outcome. We suggest that a high IGF-II/IGFBP-2 mRNA ratio may be a sign of biologically aggressive behavior in meningiomas that can influence treatment strategies. We propose that low IGFBP-2 levels in combination with increased levels of IGF-II would result in more free IGF-II and consequently greater stimulation of proliferation.
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The Human Splicing Factor ASF/SF2 can Specifically Recognize Pre-mRNA 5' Splice Sites
NASA Astrophysics Data System (ADS)
Zuo, Ping; Manley, James L.
1994-04-01
ASF/SF2 is a human protein previously shown to function in in vitro pre-mRNA splicing as an essential factor necessary for all splices and also as an alternative splicing factor, capable of switching selection of 5' splice sites. To begin to study the protein's mechanism of action, we have investigated the RNA binding properties of purified recombinant ASF/SF2. Using UV crosslinking and gel shift assays, we demonstrate that the RNA binding region of ASF/SF2 can interact with RNA in a sequence-specific manner, recognizing the 5' splice site in each of two different pre-mRNAs. Point mutations in the 5' splice site consensus can reduce binding by as much as a factor of 100, with the largest effects observed in competition assays. These findings support a model in which ASF/SF2 aids in the recognition of pre-mRNA 5' splice sites.
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Osman, Narin; Grande-Allen, K Jane; Ballinger, Mandy L; Getachew, Robel; Marasco, Silvana; O'Brien, Kevin D; Little, Peter J
2013-01-01
Calcific aortic valve disease is a progressive condition that shares some common pathogenic features with atherosclerosis. Transforming growth factor-β1 is a recognized mediator of atherosclerosis and is expressed in aortic valve lesions. Transforming growth factorβ1 stimulates glycosaminoglycan elongation of proteoglycans that is associated with increased lipid binding. We investigated the presence of transforming growth factor-β1 and downstream signaling intermediates in diseased human aortic valves and the effects of activated transforming growth factor-β1 receptor signaling on aortic valve interstitial cell proteoglycan synthesis and lipid binding as a possible mechanism for the initiation of the early lesion of calcific aortic valve disease. Diseased human aortic valve leaflets demonstrated strong immunohistochemical staining for transforming growth factor-β1 and phosphorylated Smad2/3. In primary porcine aortic valve interstitial cells, Western blots showed that transforming growth factor-β1 stimulated phosphorylation in both the carboxy and linker regions of Smad2/3, which was inhibited by the transforming growth factor-β1 receptor inhibitor SB431542. Gel electrophoresis and size exclusion chromatography demonstrated that SB431542 decreased transforming growth factor-β1-mediated [(35)S]-sulfate incorporation into proteoglycans in a dose-dependent manner. Further, in proteoglycans derived from transforming growth factor-β1-treated valve interstitial cells, gel mobility shift assays demonstrated that inhibition of transforming growth factor-β1 receptor signaling resulted in decreased lipid binding. Classic transforming growth factor-β1 signaling is present in human aortic valves in vivo and contributes to the modification of proteoglycans expressed by valve interstitial cells in vitro. These findings suggest that transforming growth factor-β1 may promote increased low-density lipoprotein binding in the early phases of calcific aortic valve disease. Copyright © 2013 Elsevier Inc. All rights reserved.
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Hamada, K; Gleason, S L; Levi, B Z; Hirschfeld, S; Appella, E; Ozato, K
1989-11-01
Transcription of major histocompatibility complex (MHC) class I genes is regulated by the conserved MHC class I regulatory element (CRE). The CRE has two factor-binding sites, region I and region II, both of which elicit enhancer function. By screening a mouse lambda gt 11 library with the CRE as a probe, we isolated a cDNA clone that encodes a protein capable of binding to region II of the CRE. This protein, H-2RIIBP (H-2 region II binding protein), bound to the native region II sequence, but not to other MHC cis-acting sequences or to mutant region II sequences, similar to the naturally occurring region II factor in mouse cells. The deduced amino acid sequence of H-2RIIBP revealed two putative zinc fingers homologous to the DNA-binding domain of steroid/thyroid hormone receptors. Although sequence similarity in other regions was minimal, H-2RIIBP has apparent modular domains characteristic of the nuclear hormone receptors. Further analyses showed that both H-2RIIBP and the natural region II factor bind to the estrogen response element (ERE) of the vitellogenin A2 gene. The ERE is composed of a palindrome, and half of this palindrome resembles the region II binding site of the MHC CRE. These results indicate that H-2RIIBP (i) is a member of the superfamily of nuclear hormone receptors and (ii) may regulate not only MHC class I genes but also genes containing the ERE and related sequences. Sequences homologous to the H-2RIIBP gene are widely conserved in the animal kingdom. H-2RIIBP mRNA is expressed in many mouse tissues, in agreement with the distribution of the natural region II factor.
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TREEGRAD: a grading program for eastern hardwoods
J.W. Stringer; D.W. Cremeans
1991-01-01
Assigning tree grades to eastern hardwoods is often a difficult task for neophyte graders. Recently several "dichotomous keys" have been developed for training graders in the USFS hardwood tree grading system. TREEGRAD uses the Tree Grading Algorithm (TGA) for determining grades from defect location data and is designed to be used as a teaching aid.
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5. Photocopy of site plan drawing, with later amendments dhowing ...
5. Photocopy of site plan drawing, with later amendments dhowing original USFS building shceduled for removal, Annex moved to present location, and addition to Warehouse (from the U.S. Forest Service, Wenatchee National Forest) Date Unkown - U.S. Forest Service Chelan Ranger Station, 428 West Woodin Avenue, Chelan, Chelan County, WA
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ERIC Educational Resources Information Center
Congress of the U.S., Washington, DC. Congressional Budget Office.
The Telecommunications Act of 1996 directs the Federal Communications Commission (FCC) to include support for advanced telecommunications--such as the Internet and computer networking--for elementary and secondary schools, public libraries, and nonprofit rural health care providers among the Universal Service Fund (USF) mandates. In its plan, the…
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Assessing the Usefulness of SAT and ACT Tests in Minority Admissions
ERIC Educational Resources Information Center
Micceri, Theodore
2010-01-01
This study sought to determine whether the use of standardized test scores contributes any useful information regarding First Time in College (FTIC) students' probable success at USF, using more detailed analysis of underrepresented minorities and women, who Micceri (2009) shows, experience substantial negative bias relative to males and whites on…
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-04-10
... DEPARTMENT OF AGRICULTURE Forest Service Lake Tahoe Basin Management Unit and Tahoe National... hereby given that the USDA Forest Service (USFS), Lake Tahoe Basin Management Unit (LTBMU), together with... reliable electrical transmission system for the north Lake Tahoe area, while accommodating currently...
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The Mammoth-June Ecosystem Management Project, Inyo National Forest
Connie Millar
1996-01-01
The Sierra Nevada Ecosystem Project (SNEP) case-study assessmentof the Mammoth-June Ecosystem Management Project(MJEMP) was undertaken to review and analyze the efficacy of alocal landscape analysis in achieving ecosystem-management objectivesin the Sierra Nevada. Of primary interest to SNEP was applicationof the new U.S. Forest Service (USFS) regional process...
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78 FR 53754 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-30
... Everglades Planning Project, Comment Period Ends: 10/15/2013, Contact: Gretchen Ehlinger 904 232-1682. EIS No. 20130251, Final EIS, USFS, MN, BWCAW Non-native Invasive Plant Management Project, Review Period Ends: 10... Project, Comment Period Ends: 10/15/2013, Contact: Pete Gomben 801-999-2182. EIS No. 20130255, Draft EIS...
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U.S. EPA, Pesticide Product Label, HOGUE AND KNOTT BLEACH, 09/26/1972
2011-04-14
... _, I' " , / I '. .t(:t. / /' / 11"" F"p,j( 11 'c, 0" '\\In (', ,( i tt·~~~d:: ... f·l I .... hln.jr. ('<1.,I,un Do not uSf' t, ,I,'! h 0 ':. i c i ear-, er ':. ! n (" 0 rl) bin at Ion \\fy! tt ...
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77 FR 49792 - Environmental Impacts Statements; Notice of Availability
Federal Register 2010, 2011, 2012, 2013, 2014
2012-08-17
....epa.gov/compliance/nepa/ . Weekly receipt of Environmental Impact Statements Filed 08/06/2012 Through..., Comment Period Ends: 10/01/2012, Contact: Brian Hasselbach 406-441-3908. EIS No. 20120266, Draft EIS, USFS... Ends: 10/01/2012, Contact: Harold Dyer 719-852-6215. EIS No. 20120267, Draft EIS, USN, VA, Outdoor...
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People, Places, and Pandas: Engaging Preschoolers with Interactive Whiteboards
ERIC Educational Resources Information Center
Berson, Ilene R.; Cross, Megan D.; Ward, Jennifer; Berson, Michael J.
2014-01-01
In this article, the authors describe a recent project undertaken at the University of South Florida's (USF) Preschool for Creative Learning. To align with the inquiry approach of their laboratory school, the environment at the Preschool is designed so that children can learn through exploration and individual initiative. The administration and…
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-05
... of National Forest System land in the Shoshone National Forest from mining in order to protect the... of National Forest System land in the Shoshone National Forest from location and entry under the... of Land Management, Interior. ACTION: Notice. SUMMARY: The United States Forest Service (USFS) has...
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76 FR 45848 - Notice of Application for Withdrawal and Public Meeting; Oregon
Federal Register 2010, 2011, 2012, 2013, 2014
2011-08-01
... approximately 5,610 acres of National Forest System lands, for a period of 5 years in aid of legislation to... period, the following described National Forest System lands from location and entry under the United... Management, Interior. ACTION: Notice. SUMMARY: The United States Forest Service (USFS) has filed an...
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76 FR 22340 - Further Inquiry Into Tribal Issues Relating to Establishment of a Mobility Fund
Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-21
... Fund (USF) to create a Mobility Fund which would employ a market-based, reverse auction mechanism to... Into Tribal Issues Relating to Establishment of a Mobility Fund AGENCY: Federal Communications... consideration by the Federal Communication Commission in connection with the proposed creation of a new Mobility...
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Second interagency conference on research in the watersheds
Kelly Elder; Kate Dwire; Robert Hubbard; Charles Rhoades; Sandra Ryan; Mike Young; Laurie Porth; Mark Dixon; Angue Goodbody
2006-01-01
The report comprises papers and abstracts from the Second Interagency Conference on Research in the Watersheds, May 16-18, Coweeta Hydrologic Laboratory, Otto NC. Authors from several agencies (e.g., USDA ARS, US-EPA, USFS) and the private sector contributed papers on topics ranging from new technology and analytical procedures, modeling, and hydrologic responses to...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-10
... application deadlines and other dates related to Auction 901 after considering comments provided in response... must file an application for support. Only after review of the application to confirm compliance with... for support to Tribal lands. 7. The USF/ICC Transformation Order established application, performance...
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Relationships between harvest of American ginseng and hardwood timber production
Stephen P. Prisley; James Chamberlain; Michael McGuffin
2012-01-01
The goal of this research was to quantify the relationship between American ginseng (Panax quinquefolius) and timber inventory and harvest. This was done through compilation and analysis of county-level data from public datasets: ginseng harvest data from U.S. Fish and Wildlife Service, US Forest Service (USFS) forest inventory and analysis (FIA)...
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DOT National Transportation Integrated Search
2016-04-18
This report for the U.S. Forest Service (USFS) documents the observations and findings of a transportation assistance group (TAG) study of Forest Road 151 on the Santa Fe National Forest near Abiquiu, New Mexico. Over the course of a three day site v...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-12
... DEPARTMENT OF AGRICULTURE Forest Service Extension of the Comment Period: The Village at Wolf Creek Access Project Draft Environmental Impact Statement AGENCY: Forest Service, USDA. ACTION... (USFS), Rio Grande National Forest announces the extension of the comment period for the Village at Wolf...
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Recreation visitor research: studies of diversity
Deborah J. Chavez; Patricia L. Winter; James D. Absher
2008-01-01
In 1987, the Pacific Southwest Research Station (PSW) of the U.S. Department of Agriculture Forest Service (USFS) chartered a research work unit to examine outdoor recreation in the wildland-urban interface. The new work unit was established to address the needs of the increasingly diverse recreation visitors to national forests. The four forest supervisors in southern...
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ERIC Educational Resources Information Center
Griffin, Melanie; Schmidt, LeEtta
2017-01-01
This article explores the implementation of special collections interlibrary loan policies and procedures at the University of South Florida (USF), focusing particularly on the development of policies related to physically loaning published materials, and traces the development of these policies through a pilot project to routinized…
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78 FR 26261 - Connect America Fund; Developing a Unified Intercarrier Compensation Regime
Federal Register 2010, 2011, 2012, 2013, 2014
2013-05-06
... acts pursuant to its delegated authority to clarify and correct certain rules in response to recent... 51.915 and 51.917, to file data with the Universal Service Administrative Company (USAC), the... consistent with the USF/ICC Transformation Order. IX. Procedural Matters A. Paperwork Reduction Act 33. This...
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A large amount of research exploring the relationship between watershed forest cover and streamflow quantity has been conducted in the southern Blue Ridge Mountains, particularly in association with the USFS Coweeta Hydrologic Laboratory and the Coweeta LTER. However, a clear ans...
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Effects of climate change on outdoor recreation [Chapter 10
Michael S. Hand; Jordan W. Smith; David L. Peterson; Nancy A. Brunswick; Carol P. Brown
2018-01-01
Federal agencies and other public land management agencies in Utah, Nevada, and southern Idaho provide and manage for numerous outdoor recreation opportunities. National forests in the U.S. Department of Agriculture Forest Service (USFS) Intermountain Region have nearly 19 million visits per year (table 10.1); adjacent National Park System units account for an...
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Reproducibility of polycarbonate reference material in toxicity evaluation
NASA Technical Reports Server (NTRS)
Hilado, C. J.; Huttlinger, P. A.
1981-01-01
A specific lot of bisphenol A polycarbonate has been used for almost four years as the reference material for the NASA-USF-PSC toxicity screening test method. The reproducibility of the test results over this period of time indicate that certain plastics may be more suitable reference materials than the more traditional cellulosic materials.
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Risk complexity and the wildland firefighter
Ivan Pupulidy
2012-01-01
Between 2000 and 2010 the US Forest Service and the Department of the Interior experienced 82 wildland fire fatalities. Our most recent organizational focus has been to eliminate fatalities. The chief of the USFS, in a letter to all employees, asked us to "suspend disbelief" with regard to the concept of a "zero fatality organization". This plea...
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Command Performance The USFS presents its annual Termiticide Report for 2014
Thomas Shelton; Donald Fye; Juliet Tang; Mark Mankowski
2015-01-01
For a termiticide to reach the American market where it's available for purchase by pest management professionals (PMPs), it must be federally registered by the U.S. Environmental Protection Agency (EPA). Chemical manufacturers wishing to register their products as a termiticide submit a registration package to the EPA containing fairly standard, nontarget...
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USDA-ARS?s Scientific Manuscript database
Tenofovir (TDF) is associated with phosphaturia and elevated 1,25 dihydroxy vitamin D (1,25-OH(2)D). Fibroblast growth factor-23 causes phosphaturia and increases in response to elevated 1,25-OH(2)D. Vitamin D binding proetin (VDBP) binds to 1,25-OH(2)D, decreasing biologic activity, and is elevated...
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USDA-ARS?s Scientific Manuscript database
BACKGROUND: Tenofovir (TDF) is associated with phosphaturia and elevated 1,25 dihydroxy vitamin D (1,25-OH(2)D). Fibroblast growth factor 23 (FGF23) causes phosphaturia and increases in response to elevated 1,25-OH(2)D. Vitamin D binding protein (VDBP) binds to 1,25-OH(2)D, decreasing its biologic...
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NASA Technical Reports Server (NTRS)
Sharina, Iraida G.; Martin, Emil; Thomas, Anthony; Uray, Karen L.; Murad, Ferid
2003-01-01
Soluble guanylyl cyclase (sGC) is a cytosolic enzyme producing the intracellular messenger cyclic guanosine monophosphate (cGMP) on activation with nitric oxide (NO). sGC is an obligatory heterodimer composed of alpha and beta subunits. We investigated human beta1 sGC transcriptional regulation in BE2 human neuroblastoma cells. The 5' upstream region of the beta1 sGC gene was isolated and analyzed for promoter activity by using luciferase reporter constructs. The transcriptional start site of the beta1 sGC gene in BE2 cells was identified. The functional significance of consensus transcriptional factor binding sites proximal to the transcriptional start site was investigated by site deletions in the 800-bp promoter fragment. The elimination of CCAAT-binding factor (CBF) and growth factor independence 1 (GFI1) binding cores significantly diminished whereas deletion of the NF1 core elevated the transcription. Electrophoretic mobility-shift assay (EMSA) and Western analysis of proteins bound to biotinated EMSA probes confirmed the interaction of GFI1, CBF, and NF1 factors with the beta1 sGC promoter. Treatment of BE2 cells with genistein, known to inhibit the CBF binding to DNA, significantly reduced protein levels of beta1 sGC by inhibiting transcription. In summary, our study represents an analysis of the human beta1 sGC promoter regulation in human neuroblastoma BE2 cells and identifies CBF as a critically important factor in beta1 sGC expression.
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MacQuarrie, Kyle L.; Yao, Zizhen; Fong, Abraham P.; Diede, Scott J.; Rudzinski, Erin R.; Hawkins, Douglas S.
2013-01-01
Rhabdomyosarcoma is a pediatric tumor of skeletal muscle that expresses the myogenic basic helix-loop-helix protein MyoD but fails to undergo terminal differentiation. Prior work has determined that DNA binding by MyoD occurs in the tumor cells, but myogenic targets fail to activate. Using MyoD chromatin immunoprecipitation coupled to high-throughput sequencing and gene expression analysis in both primary human muscle cells and RD rhabdomyosarcoma cells, we demonstrate that MyoD binds in a similar genome-wide pattern in both tumor and normal cells but binds poorly at a subset of myogenic genes that fail to activate in the tumor cells. Binding differences are found both across genomic regions and locally at specific sites that are associated with binding motifs for RUNX1, MEF2C, JDP2, and NFIC. These factors are expressed at lower levels in RD cells than muscle cells and rescue myogenesis when expressed in RD cells. MEF2C is located in a genomic region that exhibits poor MyoD binding in RD cells, whereas JDP2 exhibits local DNA hypermethylation in its promoter in both RD cells and primary tumor samples. These results demonstrate that regional and local silencing of differentiation factors contributes to the differentiation defect in rhabdomyosarcomas. PMID:23230269
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Briegel, K; Hentsch, B; Pfeuffer, I; Serfling, E
1991-01-01
The inducible, T cell-specific enhancers of murine and human Interleukin 2 (Il-2) genes contain the kB-like sequence GGGATTTCACC as an essential cis-acting enhancer motif. When cloned in multiple copies this so-called TCEd (distal T cell element) acts as an inducible proto-enhancer element in E14 T lymphoma cells, but not in HeLa cells. In extracts of induced, Il-2 secreting El4 cells three individual protein factors bind to TCEd DNA. The binding of the most prominent factor, named TCF-1 (T cell factor 1), is correlated with the proto-enhancer activity of TCEd. TCF-1 consists of two polypeptides of about 50 kD and 105 kD; the former seems to be related to the 50 kD polypeptide of NF-kB. Purified NF-kB is also able to bind to the TCEd, but TCF-1 binds stronger than NF-kB to TCEd DNA. The conversion of the TCEd to a 'perfect' NF-kB binding site leads to a tighter binding of NF-kB to TCEd DNA and, as a functional consequence, to the activity of the 'converted' TCEd motifs in HeLa cells. Thus, the substitution of the underlined A residue to a C within the GGGATTTCACC motif abolishes its T cell-restricted activity and leads to its functioning in both El4 cells and HeLa cells. These results indicate that lymphocyte-specific factors binding to the TCEd are involved in the control of T cell specific-transcription of the Il-2 gene. Images PMID:1945879
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NASA Astrophysics Data System (ADS)
Hickey-Vargas, R.; Holbik, S. P.; Ryan, J. G.; MacDonald, J. H., Jr.; Beck, M.
2015-12-01
Geoscience faculty at the University of South Florida (USF), Florida Gulf Coast University (FCGU), Valencia College (VC) and Florida International University (FIU) have teamed to construct, test and disseminate geoscience curricula in which microbeam analytical instruments are operated by undergraduates, with data gathered in the classroom in real-time over the internet. Activities have been developed for courses Physical Geology, Oceanography, Earth Materials, Mineralogy/Petrology and Stratigraphy using the Scanning Electron Microscope (SEM) and Electron Probe Microanalyzer (EPMA) housed in the Florida Center for Analytical Electron Microscopy (FCAEM; https://fcaem.fiu.edu) at FIU. Students and faculty send research materials such as polished rock sections and microfossil mounts to FCAEM to be examined during their scheduled class and lab periods. Student control of both decision-making and selection of analytical targets is encouraged. The objective of these activities is to move students from passive learning to active, self-directed inquiry at an early stage in their undergraduate career, while providing access to advanced instruments that are not available at USF, FGCU and VC. These strategies strongly facilitate student interest in undergraduate research making use of these instruments and one positive outcome to date is an increased number of students undertaking independent research projects. Prior research by USF PI Jeff Ryan indicated that various barriers related to instrument access and use hindered interested geoscience faculty in making use of these tools and strategies. In the current project, post-doctoral researcher Dr. Sven Holbik acts as a facilitator, working directly with faculty from other institutions one-on-one to provide initial training and support, including on-site visits to field check classroom technology when needed. Several new educators and institutions will initiate classroom activities using FCAEM instrumentation this Fall.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Davydova, E.K.
1987-01-01
One of the proteins participating in the process of elongation of polypeptide chains - elongation factor 2 (EF-2) - can be ADP-ribosylated at a unique amino acid residue - diphthamide. Since the ADP-ribosylation of EF-2 at dipthamide leads to a loss of affinity of the factor for RNA while the presence of RNA inhibits the ADP-ribosylation reaction, it seemed probable to the authors that diphthamide participated directly in the binding of EF-2 to DNA. The experiments presented in this article showed that this was not the case: diphthamide and the RNA-binding site are situated on different domains of EF-2. Thus,more » ADP-ribosylation of factor EF-2 in one domain leads to a loss of the ability to bind to RNA in the other. The authors investigated the mutual arrangement of diphthamide and the RNA-binding site on the EF-2 molecule by preparing a factor from rabbit reticulocytes and subjecting it to proteolytic digestion with elastase. The factor was incubated with elastase for 15 min at 37/sup 0/C at an enzyme:substrate ratio of 1:100 in buffer solution containing 20 mM Tris-HCl, pH 7.6, 10 mM KCl, 1 mM MgCl/sub 2/, and 2 mM dithiothreitol. The reaction was stopped by adding para-methylsulfonyl fluoride to 50 micro-M. The authors obtained a preparation as a result of proteolysis and applied it on a column with RNA-Sepharose and separated into two fractions: RNA-binding and without affinity for RNA. The initial preparation and its fractions were subjected to exhaustive ADP-ribosylation in the presence of diphtheria toxin and (U-/sup 14/C) nicotinaide adenine dinucleotide ((/sup 14/C)NAD) (296 mCi/mmole). The samples were analyzed electrophoretically in a polyacrylamide gel gradient in the presence of sodium dodecyl sulfate. For the detection of (/sup 14/C) ADP-ribosylated components, the gels were dried and exposed with RM-V x-ray film.« less
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Granoff, Dan M.; Giuntini, Serena; Gowans, Flor A.; Lujan, Eduardo; Sharkey, Kelsey; Beernink, Peter T.
2016-01-01
Meningococcal factor H-binding protein (FHbp) is an antigen in 2 serogroup B meningococcal vaccines. FHbp specifically binds human and some nonhuman primate complement FH. To investigate the effect of binding of FH to FHbp on protective antibody responses, we immunized infant rhesus macaques with either a control recombinant FHbp antigen that bound macaque FH or a mutant antigen with 2 amino acid substitutions and >250-fold lower affinity for FH. The mutant antigen elicited 3-fold higher serum IgG anti-FHbp titers and up to 15-fold higher serum bactericidal titers than the control FHbp vaccine. When comparing sera with similar IgG anti-FHbp titers, the antibodies elicited by the mutant antigen gave greater deposition of complement component C4b on live meningococci (classical complement pathway) and inhibited binding of FH, while the anti-FHbp antibodies elicited by the control vaccine enhanced FH binding. Thus, the mutant FHbp vaccine elicited an anti-FHbp antibody repertoire directed at FHbp epitopes within the FH binding site, which resulted in greater protective activity than the antibodies elicited by the control vaccine, which targeted FHbp epitopes outside of the FH combining site. Binding of a host protein to a vaccine antigen impairs protective antibody responses, which can be overcome with low-binding mutant antigens. PMID:27668287
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Wyhs, Nicolas; Walker, David; Giovinazzo, Hugh; Yegnasubramanian, Srinivasan; Nelson, William G
2014-08-01
Methylated DNA binding proteins such as Methyl-CpG Binding Domain Protein 2 (MBD2) can transduce DNA methylation alterations into a repressive signal by recruiting transcriptional co-repressor complexes. Interfering with MBD2 could lead to reactivation of tumor suppressor genes and therefore represents an attractive strategy for epigenetic therapy. We developed and compared fluorescence polarization (FP) and time-resolved fluorescence resonance energy transfer (TR-FRET)-based high-throughput screening (HTS) assays to identify small-molecule inhibitors of the interaction between the methyl binding domain of MBD2 (MBD2-MBD) and methylated DNA. Although both assays performed well in 96-well format, the TR-FRET assay (Z' factor = 0.58) emerged as a superior screening strategy compared with FP (Z' factor = 0.08) when evaluated in an HTS 384-well plate format. Using TR-FRET, we screened the Sigma LOPAC library for MBD2-MBD inhibitors and identified four compounds that also validated in a dose-response series. This included two known DNA intercalators (mitoxantrone and idarubicin) among two other inhibitory compounds (NF449 and aurintricarboxylic acid). All four compounds also inhibited the binding of SP-1, a transcription factor with a GC-rich binding sequence, to a methylated oligonucleotide, demonstrating that the activity was nonspecific. Our results provide proof of principle for using TR-FRET-based HTS to identify small-molecule inhibitors of MBD2 and other DNA-protein interactions. © 2014 Society for Laboratory Automation and Screening.
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Pedersen, Kim Brint; Chodavarapu, Harshita
2017-01-01
Angiotensin-converting enzyme 2 (ACE2) has protective effects on a wide range of morbidities associated with elevated angiotensin-II signaling. Most tissues, including pancreatic islets, express ACE2 mainly from the proximal promoter region. We previously found that hepatocyte nuclear factors 1α and 1β stimulate ACE2 expression from three highly conserved hepatocyte nuclear factor 1 binding motifs in the proximal promoter region. We hypothesized that other highly conserved motifs would also affect ACE2 expression. By systematic mutation of conserved elements, we identified five regions affecting ACE2 expression, of which two regions bound transcriptional activators. One of these is a functional FOXA binding motif. We further identified the main protein binding the FOXA motif in 832/13 insulinoma cells as well as in mouse pancreatic islets as FOXA2. PMID:29082356
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Saito, Motoki; Ishikawa, Fuyuki
2002-09-20
Although mammalian MBD3 contains the mCpG-binding domain (MBD) and is highly homologous with the authentic mCpG-binding protein MBD2, it was reported that the protein does not bind to mCpG specifically. Using recombinant human wild type and mutant MBD3 proteins, we demonstrated that atypical amino acids found in MBD3 MBD, namely, His-30 and Phe-34, are responsible for the inability of MBD3 to bind to mCpG. Interestingly, although H30K/F34Y MBD3 mutant protein binds to mCpG efficiently in vitro, it was not localized at the mCpG-rich pericentromeric regions in mouse cells. We also showed that Y34F MBD2b MBD, which possesses not the mCpG-specific DNA-binding activity but the nonspecific DNA-binding activity, was localized at the pericentromeric regions. These results suggested that the mCpG-specific DNA-binding activity is largely dispensable, and another factor(s) is required for the localization of MBD proteins in vivo. MBD3 was identified as a component of the NuRD/Mi2 complex that shows chromatin remodeling and histone deacetylase activities. We demonstrated that MBD3 MBD is necessary and sufficient for binding to HDAC1 and MTA2, two components of the NuRD/Mi2 complex. It was therefore suggested that mCpG-binding-defective MBD3 has evolutionarily conserved its MBD because of the secondary role played by the MBD in protein-protein interactions.
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Structural basis of gene regulation by the Grainyhead/CP2 transcription factor family
Ming, Qianqian; Roske, Yvette; Schuetz, Anja; Walentin, Katharina; Ibraimi, Ibraim; Schmidt-Ott, Kai M
2018-01-01
Abstract Grainyhead (Grh)/CP2 transcription factors are highly conserved in multicellular organisms as key regulators of epithelial differentiation, organ development and skin barrier formation. In addition, they have been implicated as being tumor suppressors in a variety of human cancers. Despite their physiological importance, little is known about their structure and DNA binding mode. Here, we report the first structural study of mammalian Grh/CP2 factors. Crystal structures of the DNA-binding domains of grainyhead-like (Grhl) 1 and Grhl2 reveal a closely similar conformation with immunoglobulin-like core. Both share a common fold with the tumor suppressor p53, but differ in important structural features. The Grhl1 DNA-binding domain binds duplex DNA containing the consensus recognition element in a dimeric arrangement, supporting parsimonious target-sequence selection through two conserved arginine residues. We elucidate the molecular basis of a cancer-related mutation in Grhl1 involving one of these arginines, which completely abrogates DNA binding in biochemical assays and transcriptional activation of a reporter gene in a human cell line. Thus, our studies establish the structural basis of DNA target-site recognition by Grh transcription factors and reveal how tumor-associated mutations inactivate Grhl proteins. They may serve as points of departure for the structure-based development of Grh/CP2 inhibitors for therapeutic applications. PMID:29309642
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Sysko, Robyn; Glasofer, Deborah R.; Hildebrandt, Tom; Klimek, Patrycja; Mitchell, James E.; Berg, Kelly C.; Peterson, Carol B.; Wonderlich, Stephen A.; Walsh, B. Timothy
2016-01-01
Objective Existing measures for DSM-IV eating disorder diagnoses have notable limitations, and there are important differences between DSM-IV and DSM-5 feeding and eating disorders. This study developed and validated a new semi-structured interview, the Eating Disorders Assessment for DSM-5 (EDA-5). Method Two studies evaluated the utility of the EDA-5. Study 1 compared the diagnostic validity of the EDA-5 to the Eating Disorder Examination (EDE) and evaluated the test-retest reliability of the new measure. Study 2 compared the diagnostic validity of an EDA-5 electronic application (“app”) to clinician interview and self-report assessments. Results In Study 1, the kappa for EDE and EDA-5 eating disorder diagnoses was 0.74 across all diagnoses (n= 64), with a range of κ=0.65 for Other Specified Feeding or Eating Disorder (OSFED)/Unspecified Feeding or Eating Disorder (USFED) to κ=0.90 for Binge Eating Disorder (BED). The EDA-5 test-retest kappa coefficient was 0.87 across diagnoses. For Study 2, clinical interview versus “app” conditions revealed a kappa of 0.83 for all eating disorder diagnoses (n=71). Across individual diagnostic categories, kappas ranged from 0.56 for OSFED/USFED to 0.94 for BN. Discussion High rates of agreement were found between diagnoses by EDA-5 and the EDE, and EDA-5 and clinical interviews. As this study supports the validity of the EDA-5 to generate DSM-5 eating disorders and the reliability of these diagnoses, the EDA-5 may be an option for the assessment of Anorexia Nervosa, Bulimia Nervosa, and BED. Additional research is needed to evaluate the utility of the EDA-5 in assessing DSM-5 feeding disorders. PMID:25639562
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Raths, S.K.
1987-01-01
Alpha-factor is a peptide of thirteen amino acids which is required for mating between the haploid mating types, a and ..cap alpha.., in Saccharomyces cerevisiae. An analogue of alpha-factor, DHP/sup 8/ DHP/sup 11/ Nle/sup 12/ tridecapeptide, was catalytically reduced in the presence of /sup 3/H gas for production of a radiolabeled pheromone suitable for use in binding studies. Incorporation of tritium resulted in /sup 3/H-alpha-factor with high specific activity, purity, biological activity and long shelf-life. Binding studies revealed that alpha-factor interacts with its receptor via a simple, reversible process which obeys the law of mass action. Association and dissociation kineticsmore » indicate values of 2.92 x 10/sup 6/ M/sup /minus/1/ min/sup -1/ for k/sub 1/ and between 4 and 7 x 10/sup /minus/2/ min/sup /minus/1/ for k/sub /minus/1/. Saturation binding studies reveal an equilibrium dissociation constant equal to 2.32 x 10/sup /minus/8/ M which approximate the kinetically-derived K/sub D/ of 2.12 x 10/sup /minus/8/ M. Scatchard and Hill analyses as well as dissociation behavior in the presence of excess unlabeled ligand indicate alpha-factor interacts with a homogeneous population of binding sites which do not interact and exhibit one affinity for the alpha-factor pheromone.« less
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Modulation of gene expression via overlapping binding sites exerted by ZNF143, Notch1 and THAP11
Ngondo-Mbongo, Richard Patryk; Myslinski, Evelyne; Aster, Jon C.; Carbon, Philippe
2013-01-01
ZNF143 is a zinc-finger protein involved in the transcriptional regulation of both coding and non-coding genes from polymerase II and III promoters. Our study deciphers the genome-wide regulatory role of ZNF143 in relation with the two previously unrelated transcription factors Notch1/ICN1 and thanatos-associated protein 11 (THAP11) in several human and murine cells. We show that two distinct motifs, SBS1 and SBS2, are associated to ZNF143-binding events in promoters of >3000 genes. Without co-occupation, these sites are also bound by Notch1/ICN1 in T-lymphoblastic leukaemia cells as well as by THAP11, a factor involved in self-renewal of embryonic stem cells. We present evidence that ICN1 binding overlaps with ZNF143 binding events at the SBS1 and SBS2 motifs, whereas the overlap occurs only at SBS2 for THAP11. We demonstrate that the three factors modulate expression of common target genes through the mutually exclusive occupation of overlapping binding sites. The model we propose predicts that the binding competition between the three factors controls biological processes such as rapid cell growth of both neoplastic and stem cells. Overall, our study establishes a novel relationship between ZNF143, THAP11 and ICN1 and reveals important insights into ZNF143-mediated gene regulation. PMID:23408857
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Mutant botrocetin-2 inhibits von Willebrand factor-induced platelet agglutination.
Matsui, T; Hori, A; Hamako, J; Matsushita, F; Ozeki, Y; Sakurai, Y; Hayakawa, M; Matsumoto, M; Fujimura, Y
2017-03-01
Essentials Botrocetin-2 (Bot2) binds to von Willebrand factor (VWF) and induces platelet agglutination. We identified Bot2 residues that are required for binding to VWF and glycoprotein (GP) Ib. We produced a mutant Bot2 that binds to VWF but inhibits platelet agglutination. Mutant Bot2 could be used as a potential anti-thrombotic reagent to block VWF-GPIb interaction. Background Botrocetin-2 (Bot2) is a botrocetin-like protein composed of α and β subunits that have been cloned from the snake Bothrops jararaca. Bot2 binds specifically to von Willebrand factor (VWF), and the complex induces glycoprotein (GP) Ib-dependent platelet agglutination. Objectives To exploit Bot2's VWF-binding capacity in order to attempt to create a mutant Bot2 that binds to VWF but inhibits platelet agglutination. Methods and Results Several point mutations were introduced into Bot2 cDNA, and the recombinant protein (recombinant Bot2 [rBot2]) was purified on an anti-botrocetin column. The mutant rBot2 with either Ala at Asp70 in the β subunit (Aspβ70Ala), or Argβ115Ala and Lysβ117Ala, showed reduced platelet agglutination-inducing activity. rBot2 with Aspβ70Ala showed little binding activity towards immobilized VWF on an ELISA plate, whereas rBot2 with Argβ115Ala/Lysβ117Ala showed reduced binding activity towards GPIb (glycocalicin) after forming a complex with VWF. rBot2 point-mutated to oppositely charged Glu at both Argβ115 and Lysβ117 showed normal binding activity towards VWF but no platelet-agglutinating activity. Furthermore, this doubly mutated protein inhibited ristocetin-induced or high shear stress-induced platelet aggregation, and restrained thrombus formation under flow conditions. Conclusions Asp70 in the β subunit of botrocetin is important for VWF binding, and Arg115 and Lys117 in the β subunit are essential for interaction with GPIb. Doubly mutated rBot2, with Argβ115Glu and Lysβ117Glu, repels GPIb and might have potential as an antithrombotic reagent that specifically blocks VWF function. This is the first report on an artificial botrocetin that can inhibit the VWF-GPIb interaction. © 2017 International Society on Thrombosis and Haemostasis.
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2011-01-01
Background The hepatitis B virus (HBV) is a major etiological factor of inflammation and damage to the liver resulting in hepatocellular carcinoma. Transcription factors play important roles in the disordered gene expression and liver injury caused by HBV. However, the molecular mechanisms behind this observation have not been defined. Results In this study, we observed that circulating prostaglandin (PGE) 2 synthesis was increased in patients with chronic hepatitis B infection, and detected elevated cyclooxygenase (COX)-2 expression in HBV- and HBx-expressing liver cells. Likewise, the association of HBx with C/EBPβ contributed to the induction of COX-2. The COX-2 promoter was hypomethylated in HBV-positive cells, and specific demethylation of CpG dinucleotides within each of the two NF-AT sites in the COX-2 promoter resulted in the increased binding affinity of NF-AT to the cognate sites in the promoter, followed by increased COX-2 expression and PGE2 accumulation. The DNA methylatransferase DNMT3B played a key role in the methylation of the COX-2 promoter, and its decreased binding to the promoter was responsible for the regional demethylation of CpG sites, and for the increased binding of transcription factors in HBV-positive cells. Conclusion Our results indicate that upregulation of COX-2 by HBV and HBx is mediated by both demethylation events and recruitment of multiple transcription factors binding to the promoter. PMID:21401943
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Heparin-binding epidermal growth factor-like growth factor promotes neuroblastoma differentiation.
Gaviglio, Angela L; Knelson, Erik H; Blobe, Gerard C
2017-05-01
High-risk neuroblastoma is characterized by undifferentiated neuroblasts and low schwannian stroma content. The tumor stroma contributes to the suppression of tumor growth by releasing soluble factors that promote neuroblast differentiation. Here we identify heparin-binding epidermal growth factor-like growth factor (HBEGF) as a potent prodifferentiating factor in neuroblastoma. HBEGF mRNA expression is decreased in human neuroblastoma tumors compared with benign tumors, with loss correlating with decreased survival. HBEGF protein is expressed only in stromal compartments of human neuroblastoma specimens, with tissue from high-stage disease containing very little stroma or HBEGF expression. In 3 human neuroblastoma cell lines (SK-N-AS, SK-N-BE2, and SH-SY5Y), soluble HBEGF is sufficient to promote neuroblast differentiation and decrease proliferation. Heparan sulfate proteoglycans and heparin derivatives further enhance HBEGF-induced differentiation by forming a complex with the epidermal growth factor receptor, leading to activation of the ERK1/2 and STAT3 pathways and up-regulation of the inhibitor of DNA binding transcription factor. These data support a role for loss of HBEGF in the neuroblastoma tumor microenvironment in neuroblastoma pathogenesis.-Gaviglio, A. L., Knelson, E. H., Blobe, G. C. Heparin-binding epidermal growth factor-like growth factor promotes neuroblastoma differentiation. © FASEB.
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Kibbey, Megan M; Jameson, Mark J; Eaton, Erin M; Rosenzweig, Steven A
2006-03-01
Signaling by the insulin-like growth factor (IGF)-1 receptor (IGF-1R) has been implicated in the promotion and aggressiveness of breast, prostate, colorectal, and lung cancers. The IGF binding proteins (IGFBPs) represent a class of natural IGF antagonists that bind to and sequester IGF-1/2 from the IGF-1R, making them attractive candidates as therapeutics for cancer prevention and control. Recombinant human IGFBP-2 significantly attenuated IGF-1-stimulated MCF-7 cell proliferation with coaddition of 20 or 100 nM IGFBP-2 (50 or 80% inhibition, respectively). We previously identified IGF-1 contact sites both upstream and downstream of the CWCV motif (residues 247-250) in human IGFBP-2 (J Biol Chem 276:2880-2889, 2001). To further test their contributions to IGFBP-2 function, the single tryptophan in human IGFBP-2, Trp-248, was selectively cleaved with 2-(2'nitrophenylsulfenyl)-3-methyl-3 bromoindolenine (BNPS-skatole) and the BNPS-skatole products IGFBP-2(1-248) and IGFBP-2(249-289) as well as IGFBP-2(1-190) were expressed as glutathione S-transferase-fusion proteins and purified. Based on competition binding analysis, deletion of residues 249 to 289 caused an approximately 20-fold decrease in IGF-1 binding affinity (IGFBP-2 EC50 = 0.35 nM and IGFBP-2(1-248) = 7 nM). Removal of the remainder of the C-terminal domain had no further effect on affinity (IGFBP-2(1-190) EC50 = 9.2 nM). In kinetic assays, IGFBP-2(1-248) and IGFBP-2(1-190) exhibited more rapid association and dissociation rates than full-length IGFBP-2. These results confirm that regions upstream and downstream of the CWCV motif participate in IGF-1 binding. They further support the development of full-length IGFBP-2 as a cancer therapeutic.
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Tan, Cheng; Li, Wenfei; Wang, Wei
2013-12-19
Protein TFIIIA is composed of nine tandemly arranged Cys2His2 zinc fingers. It can bind either to the 5S RNA gene as a transcription factor or to the 5S RNA transcript as a chaperone. Although structural and biochemical data provided valuable information on the recognition between the TFIIIIA and the 5S DNA/RNA, the involved conformational motions and energetic factors contributing to the binding affinity and specificity remain unclear. In this work, we conducted MD simulations and MM/GBSA calculations to investigate the binding-induced conformational changes in the recognition of the 5S RNA by the central three zinc fingers of TFIIIA and the energetic factors that influence the binding affinity and specificity at an atomistic level. Our results revealed drastic interdomain conformational changes between these three zinc fingers, involving the exposure/burial of several crucial DNA/RNA binding residues, which can be related to the competition between DNA and RNA for the binding of TFIIIA. We also showed that the specific recognition between finger 4/finger 6 and the 5S RNA introduces frustrations to the nonspecific interactions between finger 5 and the 5S RNA, which may be important to achieve optimal binding affinity and specificity.
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Kumar, Sunil; Ambrosini, Giovanna; Bucher, Philipp
2017-01-04
SNP2TFBS is a computational resource intended to support researchers investigating the molecular mechanisms underlying regulatory variation in the human genome. The database essentially consists of a collection of text files providing specific annotations for human single nucleotide polymorphisms (SNPs), namely whether they are predicted to abolish, create or change the affinity of one or several transcription factor (TF) binding sites. A SNP's effect on TF binding is estimated based on a position weight matrix (PWM) model for the binding specificity of the corresponding factor. These data files are regenerated at regular intervals by an automatic procedure that takes as input a reference genome, a comprehensive SNP catalogue and a collection of PWMs. SNP2TFBS is also accessible over a web interface, enabling users to view the information provided for an individual SNP, to extract SNPs based on various search criteria, to annotate uploaded sets of SNPs or to display statistics about the frequencies of binding sites affected by selected SNPs. Homepage: http://ccg.vital-it.ch/snp2tfbs/. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.
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NASA Technical Reports Server (NTRS)
Whitson, Peggy A.; Stuart, Charles A.; Huls, M. H.; Sams, Clarence F.; Cintron, Nitza M.
1989-01-01
The effect of dexamethasone on the activity of creatine kinase (CK) and the insulin-like growth factor I (IGF-I) binding were investigated using skeletal- and cardiac-muscle-derived cultured cell lines (mouse, C2C12; rat, L6 and H9c2). It was found that, in skeletal muscle cells, dexamethasone treatment during differentiation of skeletal-muscle cells caused dose-dependent increases in CK activity and increases in the degree of myotube formation, whereas cardiac cells (H9c2) exhibited very low CK activity during culture or dexamethasone treatment. Results for IGF-I binding were similar in all three cell lines. The IGF-I binding to dexamethasone-treated cells (50 nM for 24 hr on the day prior to confluence) resulted in an increased number of available binding sites, with no effect on the binding affinities.
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-04
... burden for small business concerns with fewer than 25 employees. The FCC may not conduct or sponsor a... FCC Form 525. Type of Review: Revision of a currently approved collection. Respondents: Business or... Rulemaking, 26 FCC Rcd 17663 (2011) (USF/ICC Transformation Order); see also Connect America Fund et al., WC...
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Wapiti and warblers: integrating game and nongame management in Idaho
C. R. Groves; J. W. Unsworth
1993-01-01
The primary concern of wildlife managers in the USDA Forest Service (USFS) and ldaho Department of Fish and Game (IDFG) is maintaining elk herds and quality elk hunting. As a result, nongame species like neotropical migratory landbirds do not receive much management attention. Cause for concern over this neglect are twofold: 1) forest fragmentation may be having...
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Forest Land Ownership in the Conterminous United States [map
Mark D. Nelson; Greg C. Liknes
2007-01-01
Patterns of public and private forestland ownership vary across the United States. For example, two-thirds of western forestland is publicly owned, mostly by federal agencies such as the U.S. Forest Service (USFS), Bureau of Land Management, and National Park Service. However, more than 80 percent of eastern forestland is privately owned. Private forestland is further...
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Aaron Weiskittel; Jereme Frank; David Walker; Phil Radtke; David Macfarlane; James Westfall
2015-01-01
Prediction of forest biomass and carbon is becoming important issues in the United States. However, estimating forest biomass and carbon is difficult and relies on empirically-derived regression equations. Based on recent findings from a national gap analysis and comprehensive assessment of the USDA Forest Service Forest Inventory and Analysis (USFS-FIA) component...
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Effects of climate change on nonforest vegetation [Chapter 7
Wayne G. Padgett; Matthew C. Reeves; Stanley G. Kitchen; David L. Tart; Jeanne C. Chambers; Cheri Howell; Mary E. Manning; John G. Proctor
2018-01-01
Nonforest ecosystems, as they are addressed in this chapter, contain woodland, shrubland, herbaceous, wetland, or riparian vegetation types. They are estimated to occupy over 30 million acres and 50 percent of the U.S. Department of Agriculture Forest Service (USFS) Intermountain Region (table 7.1). These diverse ecosystems range in elevation from desert floors to...
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Combining FIA plot data with topographic variables: Are precise locations needed?
Stephen P. Prisley; Huei-Jin Wang; Philip J Radtke; John Coulston
2009-01-01
Plot data from the USFS FIA program could be combined with terrain variables to attempt to explain how terrain characteristics influence forest growth, species composition, productivity, fire behavior, wildlife habitat, and other phenomena. While some types of analyses using FIA data have been shown to be insensitive to precision of plot locations, it has been...
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Linda A. Joyce; Geoffry M. Blate; Jeremy S. Littell; Steven G. McNulty; Constance I. Millar; Susanne C. Moser; Ronald P. Neilson; Kathy O' Halloran; David L. Peterson
2008-01-01
The National Forest System (NFS) is composed of 155 national forests (NFs) and 20 national grasslands (NGs), which encompass a wide range of ecosystems, harbor much of the nationâs biodiversity, and provide myriad goods and services. The mission of the U.S. Forest Service (USFS), which manages the NFS, has broadened from water and timber to sustaining ecosystem health...
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Hannah Gosnell; Nicole Robinson-Maness; Susan Charnley
2011-01-01
Unsustainable rangeland management and land conversion are significant sources of greenhouse gas emissions and global warming; however, rangelands also can be managed to mitigate climate change by enhancing carbon uptake through increased plant production and biological sequestration. According to a 2000 USFS General Technical Report, there are opportunities to make...
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DOT National Transportation Integrated Search
2007-06-27
Mountain National Forest by the interagency Transportation Assistance Group (TAG) was conducted June 27-29, 2007, on behalf of the U.S. Forest Service (USFS). This TAG report was prepared subsequent to the site visit and documents the conditions obse...
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Effects of climate change on recreation in the Northern Rockies Region [Chapter 10
Michael S. Hand; Megan Lawson
2018-01-01
Outdoor recreation is an important benefit provided by Federally managed and other public lands throughout the Rocky Mountains. National forests in the Forest Service, U.S. Department of Agriculture (USFS) Northern Region and Greater Yellowstone Area (a region hereafter called the Northern Rockies region) have an estimated 13.3 million visits per year; Yellowstone,...
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ERIC Educational Resources Information Center
Todorinova, Lily; Huse, Andy; Lewis, Barbara; Torrence, Matt
2011-01-01
Declining reference statistics, diminishing human resources, and the desire to be more proactive and embedded in academic departments, prompted the University of South Florida Library to create a taskforce for re-envisioning reference services. The taskforce was charged with examining the staffing patterns at the desk and developing…
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Jeremy S. Fried; J. Keith Gilless; Robert E. Martin
1987-01-01
The University of California's Department of Forestry and Resource Management, under contract with the California Department of Forestry and Fire Protection, has developed and released the first version of the California Fire Economics Simulator (CFES). The current release is adapted from the Initial Action Assessment component of the USFS's National Fire...
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Trends in land and water available for outdoor recreation
Lloyd C. Irland; Thomas Rumpf
1980-01-01
A data base for assessing the availability of land for outdoor recreation does not exist. Information on related issues such as vandalism, easements, and land posting is scanty. Construction of a data base for assessing land availability should be a high priority for USFS and HCRS, and for SCORP's and the RPA and RCA assessments.
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Field validation of Burned Area Reflectance Classification (BARC) products for post fire assessment
Andrew T. Hudak; Peter R. Robichaud; Jeffery B. Evans; Jess Clark; Keith Lannom; Penelope Morgan; Carter Stone
2004-01-01
The USFS Remote Sensing Applications Center (RSAC) and the USGS EROS Data Center (EDC) produce Burned Area Reflectance Classification (BARC) maps for use by Burned Area Emergency Rehabilitation (BAER) teams in rapid response to wildfires. BAER teams desire maps indicative of soil burn severity, but photosynthetic and nonphotosynthetic vegetation also influences the...
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Overview of the Future Forest Webinar Series [Chapter 1
Sarah Hines; Megan Matonis
2014-01-01
The Future Forest Webinar Series was created to facilitate dialogue between scientists and managers about the challenges and opportunities created by the mountain pine beetle1 (MPB) epidemic. A core team of scientists and managers from the USFS Rocky Mountain Research Station and the Northern and Rocky Mountain Regions worked together to develop the format and content...
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Law enforcement officers in the USDA forest service
Deborah J. Chavez; Joanne F. Tynon
2006-01-01
This paper reports results fiom the first in a series of studies evaluating perceptions of law enforcement officers (LEOs) in the US Forest Service (USFS). It is a follow-up to previous qualitative studies conducted to learn more about crime and violence in national forests and the impacts on recreation visitation and management, and test key characteristics of success...
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Rehabilitating Afghanistan's natural resources
George Hernandez
2011-01-01
The Soviet Union invaded Afghanistan in late 1979. During the next 23 years, the war between the Mujahideen Resistance and the Soviet forces, the ensuing civil war, and eventual take over by the Taliban caused enormous harm to the natural resources of Afghanistan. In 2003, the USDA Forest Service (USFS) was asked by the USDA Foreign Agricultural Service to provide...
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Michael J. Dockry; Sophia A. Gutterman; Mae A. Davenport
2017-01-01
American Indian tribes have inherent rights to national forestland and resources codified in treaties, the US Constitution, statutes, Presidential Executive Orders, and case law. These rights require a government-togovernment relationship between each tribe and the US Forest Service (USFS), which recognizes federal trust responsibilities and tribal sovereignty. This is...
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Influences on USFS District Rangers' Decision to Authorize Wildland Fire Use
Martha A. Williamson
2006-01-01
United States wildland fire policy and program reviews in 1995 and 2000 required reduction of hazardous fuel and recognition of fire as a natural process. Although an existing policy, Wildland Fire Use (WFU), permitted managing natural ignitions to meet resource benefits, most fuel reduction is still achieved through mechanical treatments and prescribed burning....
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-21
... Information Relay Service (FIRS) at 1-(800) 877-8339 to reach either of the named contacts during normal... herein by reference. The purpose of the proposed withdrawal extension is to continue the protection of... cooperative agreement would not provide adequate protection. The USFS would not need to acquire water rights...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-06
...), via fax at 202-395-5167 or via Internet at [email protected] and to Judith B. Herman, Federal Communications Commission, via the Internet at [email protected] . To submit your PRA... waste, fraud and abuse of the Universal Service Fund (USF or Fund). Among other things, the Lifeline...
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Biggar, Kyle K; Storey, Kenneth B
2018-01-01
In many cases, the DNA-binding activity of a transcription factor does not change, while its transcriptional activity is greatly influenced by the make-up of bound proteins. In this study, we assessed the protein composition and DNA-binding ability of the E2F transcription factor complex to provide insight into cell cycle control in an anoxia tolerant turtle through the use of a modified ELISA protocol. This modification also permits the use of custom DNA probes that are tailored to a specific DNA binding region, introducing the ability to design capture probes for non-model organisms. Through the use of EMSA and ELISA DNA binding assays, we have successfully determined the in vitro DNA binding activity and complex dynamics of the Rb/E2F cell cycle regulatory mechanisms in an anoxic turtle, Trachemys scripta elegans . Repressive cell cycle proteins (E2F4, Rb, HDAC4 and Suv39H1) were found to significantly increase at E2F DNA-binding sites upon anoxic exposure in anoxic turtle liver. The lack of p130 involvement in the E2F DNA-bound complex indicates that anoxic turtle liver may maintain G 1 arrest for the duration of stress survival.
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Biggar, Kyle K.
2018-01-01
In many cases, the DNA-binding activity of a transcription factor does not change, while its transcriptional activity is greatly influenced by the make-up of bound proteins. In this study, we assessed the protein composition and DNA-binding ability of the E2F transcription factor complex to provide insight into cell cycle control in an anoxia tolerant turtle through the use of a modified ELISA protocol. This modification also permits the use of custom DNA probes that are tailored to a specific DNA binding region, introducing the ability to design capture probes for non-model organisms. Through the use of EMSA and ELISA DNA binding assays, we have successfully determined the in vitro DNA binding activity and complex dynamics of the Rb/E2F cell cycle regulatory mechanisms in an anoxic turtle, Trachemys scripta elegans. Repressive cell cycle proteins (E2F4, Rb, HDAC4 and Suv39H1) were found to significantly increase at E2F DNA-binding sites upon anoxic exposure in anoxic turtle liver. The lack of p130 involvement in the E2F DNA-bound complex indicates that anoxic turtle liver may maintain G1 arrest for the duration of stress survival. PMID:29770276
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Protein-Protein Interactions within Late Pre-40S Ribosomes
Campbell, Melody G.; Karbstein, Katrin
2011-01-01
Ribosome assembly in eukaryotic organisms requires more than 200 assembly factors to facilitate and coordinate rRNA transcription, processing, and folding with the binding of the ribosomal proteins. Many of these assembly factors bind and dissociate at defined times giving rise to discrete assembly intermediates, some of which have been partially characterized with regards to their protein and RNA composition. Here, we have analyzed the protein-protein interactions between the seven assembly factors bound to late cytoplasmic pre-40S ribosomes using recombinant proteins in binding assays. Our data show that these factors form two modules: one comprising Enp1 and the export adaptor Ltv1 near the beak structure, and the second comprising the kinase Rio2, the nuclease Nob1, and a regulatory RNA binding protein Dim2/Pno1 on the front of the head. The GTPase-like Tsr1 and the universally conserved methylase Dim1 are also peripherally connected to this second module. Additionally, in an effort to further define the locations for these essential proteins, we have analyzed the interactions between these assembly factors and six ribosomal proteins: Rps0, Rps3, Rps5, Rps14, Rps15 and Rps29. Together, these results and previous RNA-protein crosslinking data allow us to propose a model for the binding sites of these seven assembly factors. Furthermore, our data show that the essential kinase Rio2 is located at the center of the pre-ribosomal particle and interacts, directly or indirectly, with every other assembly factor, as well as three ribosomal proteins required for cytoplasmic 40S maturation. These data suggest that Rio2 could play a central role in regulating cytoplasmic maturation steps. PMID:21283762
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Protein-protein interactions within late pre-40S ribosomes.
Campbell, Melody G; Karbstein, Katrin
2011-01-20
Ribosome assembly in eukaryotic organisms requires more than 200 assembly factors to facilitate and coordinate rRNA transcription, processing, and folding with the binding of the ribosomal proteins. Many of these assembly factors bind and dissociate at defined times giving rise to discrete assembly intermediates, some of which have been partially characterized with regards to their protein and RNA composition. Here, we have analyzed the protein-protein interactions between the seven assembly factors bound to late cytoplasmic pre-40S ribosomes using recombinant proteins in binding assays. Our data show that these factors form two modules: one comprising Enp1 and the export adaptor Ltv1 near the beak structure, and the second comprising the kinase Rio2, the nuclease Nob1, and a regulatory RNA binding protein Dim2/Pno1 on the front of the head. The GTPase-like Tsr1 and the universally conserved methylase Dim1 are also peripherally connected to this second module. Additionally, in an effort to further define the locations for these essential proteins, we have analyzed the interactions between these assembly factors and six ribosomal proteins: Rps0, Rps3, Rps5, Rps14, Rps15 and Rps29. Together, these results and previous RNA-protein crosslinking data allow us to propose a model for the binding sites of these seven assembly factors. Furthermore, our data show that the essential kinase Rio2 is located at the center of the pre-ribosomal particle and interacts, directly or indirectly, with every other assembly factor, as well as three ribosomal proteins required for cytoplasmic 40S maturation. These data suggest that Rio2 could play a central role in regulating cytoplasmic maturation steps.
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Brabletz, T; Pietrowski, I; Serfling, E
1991-01-01
Like Cyclosporin A (CsA), the macrolide FK 506 is a potent immunosuppressive that inhibits early steps of T cell activation, including the synthesis of Interleukin 2 (II-2) and numerous other lymphokines. The block of II-2 synthesis occurs at the transcriptional level. At concentrations that block T cell activation, FK 506 and CsA inhibit the proto-enhancer activity of Purine boxes of the II-2 promoter and the generation of lymphocyte-specific factors binding to the Purine boxes. Under the same conditions, the DNA binding of other II-2 enhancer factors remains unaffected by both compounds. These results support the view that FK 506 and CsA, which both inhibit the activity of peptidylprolyl cis/trans isomerases, suppress T cell activation by a similar, if not identical mechanism. Images PMID:1707162
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Brabletz, T; Pietrowski, I; Serfling, E
1991-01-11
Like Cyclosporin A (CsA), the macrolide FK 506 is a potent immunosuppressive that inhibits early steps of T cell activation, including the synthesis of Interleukin 2 (II-2) and numerous other lymphokines. The block of II-2 synthesis occurs at the transcriptional level. At concentrations that block T cell activation, FK 506 and CsA inhibit the proto-enhancer activity of Purine boxes of the II-2 promoter and the generation of lymphocyte-specific factors binding to the Purine boxes. Under the same conditions, the DNA binding of other II-2 enhancer factors remains unaffected by both compounds. These results support the view that FK 506 and CsA, which both inhibit the activity of peptidylprolyl cis/trans isomerases, suppress T cell activation by a similar, if not identical mechanism.
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The pig CYP2E1 promoter is activated by COUP-TF1 and HNF-1 and is inhibited by androstenone.
Tambyrajah, Winston S; Doran, Elena; Wood, Jeffrey D; McGivan, John D
2004-11-15
Functional analysis of the pig cytochrome P4502E1 (CYP2E1) promoter identified two major activating elements. One corresponded to the hepatic nuclear factor 1 (HNF-1) consensus binding sequence at nucleotides -128/-98 and the other was located in the region -292/-266. The binding of proteins in pig liver nuclear extracts to a synthetic double-stranded oligonucleotide corresponding to this more distal activating sequence was studied by electrophoretic mobility shift assay. The minimum protein binding sequence was identified as TGTTCTGACCTCTGGG. Gel super-shift assays identified the protein binding to this site as chick ovalbumin upstream promoter transcription factor 1 (COUP-TF1). Androstenone inhibited promoter activity in transfection experiments only with constructs which included the COUP-TF1 binding site. Androstenone inhibited COUP-TF1 binding to synthetic oligonucleotides but did not affect HNF-1 binding. The results offer an explanation for the inhibition of CYP2E1 protein expression by androstenone in isolated pig hepatocytes and may be relevant to the low expression of hepatic CYP2E1 in those pigs which accumulate high levels of androstenone in vivo.
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Selective Activation of Transcription by a Novel CCAAT Binding Factor
NASA Astrophysics Data System (ADS)
Maity, Sankar N.; Golumbek, Paul T.; Karsenty, Gerard; de Crombrugghe, Benoit
1988-07-01
A novel CCAAT binding factor (CBF) composed of two different subunits has been extensively purified from rat liver. Both subunits are needed for specific binding to DNA. Addition of this purified protein to nuclear extracts of NIH 3T3 fibroblasts stimulates transcription from several promoters including the α 2(I) collagen, the α 1(I) collagen, the Rous sarcoma virus long terminal repeat (RSV-LTR), and the adenovirus major late promoter. Point mutations in the CCAAT motif that show either no binding or a decreased binding of CBF likewise abolish or reduce activation of transcription by CBF. Activation of transcription requires, therefore, the specific binding of CBF to its recognition sites.
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Zamora, Paul O [Gaithersburg, MD; Pena, Louis A [Poquott, NY; Lin, Xinhua [Plainview, NY; Takahashi, Kazuyuki [Germantown, MD
2012-07-24
The present invention provides a fibroblast growth factor heparin-binding analog of the formula: ##STR00001## where R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, X, Y and Z are as defined, pharmaceutical compositions, coating compositions and medical devices including the fibroblast growth factor heparin-binding analog of the foregoing formula, and methods and uses thereof.
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Heparin-binding epidermal growth factor-like growth factor promotes neuroblastoma differentiation
Gaviglio, Angela L.; Knelson, Erik H.; Blobe, Gerard C.
2017-01-01
High-risk neuroblastoma is characterized by undifferentiated neuroblasts and low schwannian stroma content. The tumor stroma contributes to the suppression of tumor growth by releasing soluble factors that promote neuroblast differentiation. Here we identify heparin-binding epidermal growth factor–like growth factor (HBEGF) as a potent prodifferentiating factor in neuroblastoma. HBEGF mRNA expression is decreased in human neuroblastoma tumors compared with benign tumors, with loss correlating with decreased survival. HBEGF protein is expressed only in stromal compartments of human neuroblastoma specimens, with tissue from high-stage disease containing very little stroma or HBEGF expression. In 3 human neuroblastoma cell lines (SK-N-AS, SK-N-BE2, and SH-SY5Y), soluble HBEGF is sufficient to promote neuroblast differentiation and decrease proliferation. Heparan sulfate proteoglycans and heparin derivatives further enhance HBEGF-induced differentiation by forming a complex with the epidermal growth factor receptor, leading to activation of the ERK1/2 and STAT3 pathways and up-regulation of the inhibitor of DNA binding transcription factor. These data support a role for loss of HBEGF in the neuroblastoma tumor microenvironment in neuroblastoma pathogenesis.—Gaviglio, A. L., Knelson, E. H., Blobe, G. C. Heparin-binding epidermal growth factor-like growth factor promotes neuroblastoma differentiation. PMID:28174207
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Structural and functional analysis of the YAP-binding domain of human TEAD2
DOE Office of Scientific and Technical Information (OSTI.GOV)
Tian, Wei; Yu, Jianzhong; Tomchick, Diana R.
2010-06-15
The Hippo pathway controls organ size and suppresses tumorigenesis in metazoans by blocking cell proliferation and promoting apoptosis. The TEAD1-4 proteins (which contain a DNA-binding domain but lack an activation domain) interact with YAP (which lacks a DNA-binding domain but contains an activation domain) to form functional heterodimeric transcription factors that activate proliferative and prosurvival gene expression programs. The Hippo pathway inhibits the YAP-TEAD hybrid transcription factors by phosphorylating and promoting cytoplasmic retention of YAP. Here we report the crystal structure of the YAP-binding domain (YBD) of human TEAD2. TEAD2 YBD adopts an immunoglobulin-like {beta}-sandwich fold with two extra helix-turn-helixmore » inserts. NMR studies reveal that the TEAD-binding domain of YAP is natively unfolded and that TEAD binding causes localized conformational changes in YAP. In vitro binding and in vivo functional assays define an extensive conserved surface of TEAD2 YBD as the YAP-binding site. Therefore, our studies suggest that a short segment of YAP adopts an extended conformation and forms extensive contacts with a rigid surface of TEAD. Targeting a surface-exposed pocket of TEAD might be an effective strategy to disrupt the YAP-TEAD interaction and to reduce the oncogenic potential of YAP.« less
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Yasuhiko, Yukuto; Kitajima, Satoshi; Takahashi, Yu; Oginuma, Masayuki; Kagiwada, Harumi; Kanno, Jun; Saga, Yumiko
2008-11-01
The T-box transcription factor Tbx6 controls the expression of Mesp2, which encodes a basic helix-loop-helix transcription factor that has crucial roles in somitogenesis. In cultured cells, Tbx6 binding to the Mesp2 enhancer region is essential for the activation of Mesp2 by Notch signaling. However, it is not known whether this binding is required in vivo. Here we report that an Mesp2 enhancer knockout mouse bearing mutations in two crucial Tbx6 binding sites does not express Mesp2 in the presomitic mesoderm. This absence leads to impaired skeletal segmentation identical to that reported for Mesp2-null mice, indicating that these Tbx6 binding sites are indispensable for Mesp2 expression. T-box binding to the consensus sequences in the Mesp2 upstream region was confirmed by chromatin immunoprecipitation assays. Further enhancer analyses indicated that the number and spatial organization of the T-box binding sites are critical for initiating Mesp2 transcription via Notch signaling. We also generated a knock-in mouse in which the endogenous Mesp2 enhancer was replaced by the core enhancer of medaka mespb, an ortholog of mouse Mesp2. The homozygous enhancer knock-in mouse was viable and showed normal skeletal segmentation, indicating that the medaka mespb enhancer functionally replaced the mouse Mesp2 enhancer. These results demonstrate that there is significant evolutionary conservation of Mesp regulatory mechanisms between fish and mice.
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Signaling by Kit protein-tyrosine kinase--the stem cell factor receptor.
Roskoski, Robert
2005-11-11
Signaling by stem cell factor and Kit, its receptor, plays important roles in gametogenesis, hematopoiesis, mast cell development and function, and melanogenesis. Moreover, human and mouse embryonic stem cells express Kit transcripts. Stem cell factor exists as both a soluble and a membrane-bound glycoprotein while Kit is a receptor protein-tyrosine kinase. The complete absence of stem cell factor or Kit is lethal. Deficiencies of either produce defects in red and white blood cell production, hypopigmentation, and sterility. Gain-of-function mutations of Kit are associated with several human neoplasms including acute myelogenous leukemia, gastrointestinal stromal tumors, and mastocytomas. Kit consists of an extracellular domain, a transmembrane segment, a juxtamembrane segment, and a protein kinase domain that contains an insert of about 80 amino acid residues. Binding of stem cell factor to Kit results in receptor dimerization and activation of protein kinase activity. The activated receptor becomes autophosphorylated at tyrosine residues that serve as docking sites for signal transduction molecules containing SH2 domains. The adaptor protein APS, Src family kinases, and Shp2 tyrosyl phosphatase bind to phosphotyrosine 568. Shp1 tyrosyl phosphatase and the adaptor protein Shc bind to phosphotyrosine 570. C-terminal Src kinase homologous kinase and the adaptor Shc bind to both phosphotyrosines 568 and 570. These residues occur in the juxtamembrane segment of Kit. Three residues in the kinase insert domain are phosphorylated and attract the adaptor protein Grb2 (Tyr703), phosphatidylinositol 3-kinase (Tyr721), and phospholipase Cgamma (Tyr730). Phosphotyrosine 900 in the distal kinase domain binds phosphatidylinositol 3-kinase which in turn binds the adaptor protein Crk. Phosphotyrosine 936, also in the distal kinase domain, binds the adaptor proteins APS, Grb2, and Grb7. Kit has the potential to participate in multiple signal transduction pathways as a result of interaction with several enzymes and adaptor proteins.
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Binding and Translocation of Termination Factor Rho Studied at the Single-Molecule Level
Koslover, Daniel J.; Fazal, Furqan M.; Mooney, Rachel A.; Landick, Robert; Block, Steven M.
2012-01-01
Rho termination factor is an essential hexameric helicase responsible for terminating 20–50% of all mRNA synthesis in E. coli. We used single- molecule force spectroscopy to investigate Rho-RNA binding interactions at the Rho- utilization (rut) site of the ? tR1 terminator. Our results are consistent with Rho complexes adopting two states, one that binds 57 ±2 nucleotides of RNA across all six of the Rho primary binding sites, and another that binds 85 ±2 nucleotides at the six primary sites plus a single secondary site situated at the center of the hexamer. The single-molecule data serve to establish that Rho translocates 5′-to-3′ towards RNA polymerase (RNAP) by a tethered-tracking mechanism, looping out the intervening RNA between the rut site and RNAP. These findings lead to a general model for Rho binding and translocation, and establish a novel experimental approach that should facilitate additional single- molecule studies of RNA-binding proteins. PMID:22885804
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Selective inhibition of c-Myc/Max dimerization and DNA binding by small molecules.
Kiessling, Anke; Sperl, Bianca; Hollis, Angela; Eick, Dirk; Berg, Thorsten
2006-07-01
bZip and bHLHZip protein family members comprise a large fraction of eukaryotic transcription factors and need to bind DNA in order to exert most of their fundamental biological roles. Their binding to DNA requires homo- or heterodimerization via alpha-helical domains, which generally do not contain obvious binding sites for small molecules. We have identified two small molecules, dubbed Mycro1 and Mycro2, which inhibit the protein-protein interactions between the bHLHZip proteins c-Myc and Max. Mycros are the first inhibitors of c-Myc/Max dimerization, which have been demonstrated to inhibit DNA binding of c-Myc with preference over other dimeric transcription factors in vitro. Mycros inhibit c-Myc-dependent proliferation, gene transcription, and oncogenic transformation in the low micromolar concentration range. Our data support the idea that dimeric transcription factors can be druggable even in the absence of obvious small-molecule binding pockets.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Buchman, A.R.; Kimmerly, W.J.; Rine, J.
1988-01-01
Two DNA-binding factors from Saccharomyces cerevisiae have been characterized, GRFI (general regulatory factor I) and ABFI (ARS-binding factor I), that recognize specific sequences within diverse genetic elements. GRFI bound to sequences at the negative regulatory elements (silencers) of the silent mating type loci HML E and HMR E and to the upstream activating sequence (UAS) required for transcription of the MAT ..cap alpha.. genes. A putative conserved UAS located at genes involved in translation (RPG box) was also recognized by GRFI. In addition, GRFI bound with high affinity to sequences within the (C/sub 1-3/A)-repeat region at yeast telomeres. Binding sitesmore » for GRFI with the highest affinity appeared to be of the form 5'-(A/G)(A/C)ACCCAN NCA(T/C)(T/C)-3', where N is any nucleotide. ABFI-binding sites were located next to autonomously replicating sequences (ARSs) at controlling elements of the silent mating type loci HMR E, HMR I, and HML I and were associated with ARS1, ARS2, and the 2..mu..m plasmid ARS. Two tandem ABFI binding sites were found between the HIS3 and DED1 genes, several kilobase pairs from any ARS, indicating that ABFI-binding sites are not restricted to ARSs. The sequences recognized by AFBI showed partial dyad-symmetry and appeared to be variations of the consensus 5'-TATCATTNNNNACGA-3'. GRFI and ABFI were both abundant DNA-binding factors and did not appear to be encoded by the SIR genes, whose product are required for repression of the silent mating type loci. Together, these results indicate that both GRFI and ABFI play multiple roles within the cell.« less
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Stockbauer, K E; Magoun, L; Liu, M; Burns, E H; Gubba, S; Renish, S; Pan, X; Bodary, S C; Baker, E; Coburn, J; Leong, J M; Musser, J M
1999-01-05
The human pathogenic bacterium group A Streptococcus produces an extracellular cysteine protease [streptococcal pyrogenic exotoxin B (SpeB)] that is a critical virulence factor for invasive disease episodes. Sequence analysis of the speB gene from 200 group A Streptococcus isolates collected worldwide identified three main mature SpeB (mSpeB) variants. One of these variants (mSpeB2) contains an Arg-Gly-Asp (RGD) sequence, a tripeptide motif that is commonly recognized by integrin receptors. mSpeB2 is made by all isolates of the unusually virulent serotype M1 and several other geographically widespread clones that frequently cause invasive infections. Only the mSpeB2 variant bound to transfected cells expressing integrin alphavbeta3 (also known as the vitronectin receptor) or alphaIIbbeta3 (platelet glycoprotein IIb-IIIa), and binding was blocked by a mAb that recognizes the streptococcal protease RGD motif region. In addition, mSpeB2 bound purified platelet integrin alphaIIbbeta3. Defined beta3 mutants that are altered for fibrinogen binding were defective for SpeB binding. Synthetic peptides with the mSpeB2 RGD motif, but not the RSD sequence present in other mSpeB variants, blocked binding of mSpeB2 to transfected cells expressing alphavbeta3 and caused detachment of cultured human umbilical vein endothelial cells. The results (i) identify a Gram-positive virulence factor that directly binds integrins, (ii) identify naturally occurring variants of a documented Gram-positive virulence factor with biomedically relevant differences in their interactions with host cells, and (iii) add to the theme that subtle natural variation in microbial virulence factor structure alters the character of host-pathogen interactions.
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Characterization of insulin-like growth factor I receptor on human erythrocytes.
Hizuka, N; Takano, K; Tanaka, I; Honda, N; Tsushima, T; Shizume, K
1985-12-01
[125I]Insulin-like growth factor I (IGF-I) specifically bound to erythrocytes; the binding was saturable, and time and temperature dependent. Steady state binding was reached at 16 h at 4 C, and specific binding averaged 14.3 +/- 0.7% (+/- SEM) at a concentration of 3.6 X 10(9) cells/ml in seven normal subjects. [125I]IGF-I binding to the cells was displaced by unlabeled IGF-I in a dose-dependent manner. Scatchard analysis indicated a linear plot, and Ka and number of binding sites/cell were 1.43 +/- 0.07 X 10(9) M-1 and 20.7 +/- 2.2, respectively. Compared to IGF-I, the relative potencies of multiplication-stimulating activity and insulin for displacing [125I]IGF-I binding were 20% and 1%, respectively. [125I]IGF-I binding to erythrocytes from patients with acromegaly was lower than binding to cells from pituitary dwarfs. An inverse correlation between plasma IGF-I level and the number of IGF-I-binding sites per cell was found (r = -0.75; P less than 0.005). These results demonstrate that [125I]IGF-I binding to erythrocytes can be used for clinical measurement of the IGF-I receptor.
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Jayanthi, Srinivas; Kathir, Karuppanan Muthusamy; Rajalingam, Dakshinamurthy; Furr, Mercede; Daily, Anna; Thurman, Ryan; Rutherford, Lindsay; Chandrashekar, Reena; Adams, Paul; Prudovsky, Igor; Suresh Kumar, Thallapuranam Krishnaswamy
2014-01-01
Fibroblast growth factor 1 (FGF1) is a heparin-binding proangiogenic protein. FGF1 lacks the conventional N-terminal signal peptide required for secretion through the endoplasmic reticulum (ER) -Golgi secretory pathway. FGF1 is released through a Cu2+ - mediated nonclassical secretion pathway. The secretion of FGF1 involves the formation of a Cu2+- mediated multiprotein release complex (MRC) including FGF1, S100A13 (a calcium-binding protein) and p40 synaptotagmin (Syt1). It is believed that binding of Cu2+ to the C2B domain is important for the release of FGF1 in to the extracellular medium. In this study, using a variety of biophysical studies, Cu2+ and lipid interactions of the C2B domain of Syt1were characterized. Isothermal titration calorimetry (ITC) experiments reveal that C2B domain binds to Cu2+ in a biphasic manner involving an initial endothermic and a subsequent exothermic phase. Fluorescence energy transfer experiments using Tb3+ show that there are two Cu2+- binding pockets on the C2B domain, and one of these is also a Ca2+- binding site. Lipid-binding studies using ITC demonstrate that the C2B domain preferentially binds to small unilamellar vesicles of phosphatidyl serine (PS). Results of the differential scanning calorimetry and limited trypsin digestion experiments suggest that C2B domain is marginally destabilized upon binding to PS vesicles. These results, for the first time, suggest that the main role of the C2B domain of Syt1 is to serve as an anchor for the FGF1 MRC on the membrane bilayer. In addition, binding of the C2B domain to the lipid bilayer is shown to significantly decrease the binding affinity of the protein to Cu2+. The study provides valuable insights on the sequence of structural events that occur in the nonclassical secretion of FGF1. PMID:25224745
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Zhao, Jinshan; Li, Hegang; Liu, Kaidong; Zhang, Baoxun; Li, Peipei; He, Jianning; Cheng, Ming; De, Wei; Liu, Jifeng; Zhao, Yaofeng; Yang, Lihua; Liu, Nan
2016-10-01
Goats are an important source of fibers. In the present study microarray technology was used to investigate the potential genes primarily involved in hair and cashmere growth in the Laiwu black goat. A total of 655 genes differentially expressed in body (hair‑growing) and groin (hairless) skin were identified, and their potential association with hair and cashmere growth was analyzed. The majority of genes associated with hair growth regulation could be assigned to intracellular, intracellular organelle, membrane‑bound vesicle, cytoplasmic vesicle, pattern binding, heparin binding, polysaccharide binding, glycosaminoglycan binding and cytoplasmic membrane‑bound vesicle categories. Numerous genes upregulated in body compared with groin skin contained common motifs for nuclear factor 1A, Yi, E2 factor (E2F) and cyclic adenosine monophosphate response element binding (CREB)/CREBβ binding sites in their promoter region. The promoter region of certain genes downregulated in body compared with groin skin contained three common regions with LF‑A1, Yi, E2F, Collier/Olfactory‑1/early B‑cell factor 1, peroxisome proliferator‑activated receptor α or U sites. Thus, the present study identified molecules in the cashmere‑bearing skin area of the Laiwu black goat, which may contribute to hair and cashmere traits.
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Dai, Hanjun; Umarov, Ramzan; Kuwahara, Hiroyuki; Li, Yu; Song, Le; Gao, Xin
2017-11-15
An accurate characterization of transcription factor (TF)-DNA affinity landscape is crucial to a quantitative understanding of the molecular mechanisms underpinning endogenous gene regulation. While recent advances in biotechnology have brought the opportunity for building binding affinity prediction methods, the accurate characterization of TF-DNA binding affinity landscape still remains a challenging problem. Here we propose a novel sequence embedding approach for modeling the transcription factor binding affinity landscape. Our method represents DNA binding sequences as a hidden Markov model which captures both position specific information and long-range dependency in the sequence. A cornerstone of our method is a novel message passing-like embedding algorithm, called Sequence2Vec, which maps these hidden Markov models into a common nonlinear feature space and uses these embedded features to build a predictive model. Our method is a novel combination of the strength of probabilistic graphical models, feature space embedding and deep learning. We conducted comprehensive experiments on over 90 large-scale TF-DNA datasets which were measured by different high-throughput experimental technologies. Sequence2Vec outperforms alternative machine learning methods as well as the state-of-the-art binding affinity prediction methods. Our program is freely available at https://github.com/ramzan1990/sequence2vec. xin.gao@kaust.edu.sa or lsong@cc.gatech.edu. Supplementary data are available at Bioinformatics online. © The Author(s) 2017. Published by Oxford University Press.
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Baumann, G; Geisse, S; Sullivan, M
1991-03-01
The structurally unrelated immunosuppressive drugs cyclosporin A (Sandimmun) and FK-506 both interfere with the process of T-cell proliferation by blocking the transcription of the T-cell growth factor interleukin-2 (IL-2). Here we demonstrate that the transcriptional activation of this gene requires the binding of regulatory nuclear proteins to a promoter element with sequence similarity to the consensus binding site for NF-kappa B-related transcription factors. We present evidence that the binding by regulatory nuclear proteins to the kappa B element of the IL-2 promoter is affected negatively by cyclosporin A and FK-506 at concentrations paralleling their immunosuppressive activity in vivo. The decrease in DNA-protein complex formation induced by the immunosuppressive drugs correlates with a decrease in IL-2 production. FK-506 is 10 to 100 times more potent than cyclosporin A in its ability to inhibit sequence-specific DNA binding and IL-2 production. Our findings suggest that the actions of both drugs converge at the level of DNA-protein interaction.
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Ectoderm gene activation in sea urchin embryos mediated by the CCAAT-binding factor.
Li, Xiaotao; Bhattacharya, Chitralekha; Dayal, Sandeep; Maity, Sankar; Klein, William H
2002-05-01
Transcriptional enhancers are short stretches of DNA that function to achieve highly specific patterns of gene expression. To identify the mechanisms by which enhancers achieve their specificity, we made use of an enhancer from the aboral ectoderm-specific spec2a gene of the sea urchin Strongylocentrotus purpuratus. The spec2a enhancer contains five cis-regulatory elements within 78 base pairs that interact with five distinct DNA-binding proteins to confer aboral ectoderm expression. Here, we present an analysis of the sea urchin CCAAT binding factor (CBF), which binds to a CCAAT motif within the spec2a enhancer. S. purpuratus CBF and SpOtx, a ubiquitously expressed factor, act together at closely placed cis-regulatory elements to mediate spec2a transcription in the ectoderm. SpCBF was the sole factor that bound to the spec2a CCAAT element, and two of the three subunits that make up the CBF holoprotein were cloned and shown to have high sequence conservation with their vertebrate orthologs. Based on its involvement in the regulation of several other sea urchin genes, SpCBF appears to be a major transcription factor in the sea urchin embryo for positive regulation of ectoderm gene expression. In addition to its role in vertebrate cell growth and proliferation, our results indicate that CBF also functions at the early stages of germ layer formation, namely ectoderm differentiation.
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Larabee, Jason L; Hocker, James R; Hanas, Jay S
2009-03-01
The anti-inflammatory selenium compounds, ebselen (2-phenyl-1,2-benzisoselenazol-3[2H]-one) and selenite, were found to alter the DNA binding mechanisms and structures of cysteine-rich zinc-finger transcription factors. As assayed by DNase I protection, DNA binding by TFIIIA (transcription factor IIIA, prototypical Cys(2)His(2) zinc finger protein), was inhibited by micromolar amounts of ebselen. In a gel shift assay, ebselen inhibited the Cys(2)His(2) zinc finger-containing DNA binding domain (DBD) of the NF-kappaB mediated transcription factor Sp1. Ebselen also inhibited DNA binding by the p50 subunit of the pro-inflammatory Cys-containing NF-kappaB transcription factor. Electrospray ionization mass spectrometry (ESI-MS) was utilized to elucidate mechanisms of chemical interaction between ebselen and a zinc-bound Cys(2)His(2) zinc finger polypeptide modeled after the third finger of Sp1 (Sp1-3). Exposing Sp1-3 to micromolar amounts of ebselen resulted in Zn(2+) release from this peptide and the formation of a disulfide bond by oxidation of zinc finger SH groups, the likely mechanism for DNA binding inhibition. Selenite was shown by ESI-MS to also eject zinc from Sp1-3 as well as induce disulfide bond formation through SH oxidation. The selenite-dependent inhibition/oxidation mechanism differed from that of ebselen by inducing the formation of a stable selenotrisulfide bond. Selenite-induced selenotrisulfide formation was dependent upon the structure of the Cys(2)His(2) zinc finger as alteration in the finger structure enhanced this reaction as well as selenite-dependent zinc release. Ebselen and selenite-dependent inhibition/oxidation of Cys-rich zinc finger proteins, with concomitant release of zinc and finger structural changes, points to mechanisms at the atomic and protein level for selenium-induced alterations in Cys-rich proteins, and possible amelioration of certain inflammatory, neurodegenerative, and oncogenic responses.
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Core-binding factor beta interacts with Runx2 and is required for skeletal development.
Yoshida, Carolina A; Furuichi, Tatsuya; Fujita, Takashi; Fukuyama, Ryo; Kanatani, Naoko; Kobayashi, Shinji; Satake, Masanobu; Takada, Kenji; Komori, Toshihisa
2002-12-01
Core-binding factor beta (CBFbeta, also called polyomavirus enhancer binding protein 2beta (PEBP2B)) is associated with an inversion of chromosome 16 and is associated with acute myeloid leukemia in humans. CBFbeta forms a heterodimer with RUNX1 (runt-related transcription factor 1), which has a DNA binding domain homologous to the pair-rule protein runt in Drosophila melanogaster. Both RUNX1 and CBFbeta are essential for hematopoiesis. Haploinsufficiency of another runt-related protein, RUNX2 (also called CBFA1), causes cleidocranial dysplasia in humans and is essential in skeletal development by regulating osteoblast differentiation and chondrocyte maturation. Mice deficient in Cbfb (Cbfb(-/-)) die at midgestation, so the function of Cbfbeta in skeletal development has yet to be ascertained. To investigate this issue, we rescued hematopoiesis of Cbfb(-/-) mice by introducing Cbfb using the Gata1 promoter. The rescued Cbfb(-/-) mice recapitulated fetal liver hematopoiesis in erythroid and megakaryocytic lineages and survived until birth, but showed severely delayed bone formation. Although mesenchymal cells differentiated into immature osteoblasts, intramembranous bones were poorly formed. The maturation of chondrocytes into hypertrophic cells was markedly delayed, and no endochondral bones were formed. Electrophoretic mobility shift assays and reporter assays showed that Cbfbeta was necessary for the efficient DNA binding of Runx2 and for Runx2-dependent transcriptional activation. These findings indicate that Cbfbeta is required for the function of Runx2 in skeletal development.
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The complete sequence and promoter activity of the human A-raf-1 gene (ARAF1)
DOE Office of Scientific and Technical Information (OSTI.GOV)
Lee, J.E.; Beck, T.W.; Brennscheidt, U.
1994-03-01
The raf proto-oncogenes encode cytoplasmic protein serine/threonine kinases, which play a critical role in cell growth and development. One of these, A-raf-1 (human gene symbol, ARAF1), which is predominantly expressed in mouse urogenital tissues, has been mapped to an evolutionarily conserved linkage group composed of ARAF1, SYN1, TIMP, and properdin located at human chromosome Xp11.2. The authors have isolated human genomic DNA clones containing the expressed gene (ARAF1) on the X chromosome and a pseudogene (ARAF2) on chromosome 7p12-q11.21. Analysis of the nucleotide sequence from the ARAF1 genomic clones demonstrated that it consists of 16 exons encoded by minimally 10,776more » nucleotides. The major transcriptional start site (+1) was determined by RNase protection and primer extension assays. Promoter activity was confirmed by functional assays using DNA fragments fused to a CAT reporter gene. The ARAF1 minimal promoter, located between nucleotides -59 and +93, has a low G + C content and lacks consensus TATA and Inr sequences but shows sequence similarity at position -1 to the E box that is known to interact with USF and TFII-I transcription factors. 65 refs., 7 figs., 1 tab.« less
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Hyrsova, Lucie; Smutny, Tomas; Carazo, Alejandro; Moravcik, Stefan; Mandikova, Jana; Trejtnar, Frantisek; Gerbal‐Chaloin, Sabine
2016-01-01
Background and Purpose The organic cation transporter 1 (OCT1) transports cationic drugs into hepatocytes. The high hepatic expression of OCT1 is controlled by the HNF4α and USF transcription factors. Pregnane X receptor (PXR) mediates induction of the principal xenobiotic metabolizing enzymes and transporters in the liver. Here, we have assessed the down‐regulation of OCT1 expression by PXR activation. Experimental Approach We used primary human hepatocytes and related cell lines to measure OCT1 expression and activity, by assaying MPP+ accumulation. Western blotting, qRT‐PCR, the OCT1 promoter gene reporter constructs and chromatin immunoprecipitation assays were also used. Key Results OCT1 mRNA in human hepatocytes was down‐regulated along with reduced [3H]MPP+ accumulation in differentiated HepaRG cells after treatment with rifampicin. Rifampicin and hyperforin as well as the constitutively active PXR mutant T248D suppressed activity of the 1.8 kb OCT1 promoter construct in gene reporter assays. Silencing of both PXR and HNF4α in HepaRG cells blocked the PXR ligand‐mediated down‐regulation of OCT1 expression. The mutation of HNF4α and USF1 (E‐box) responsive elements reversed the PXR‐mediated inhibition in gene reporter assays. Chromatin immunoprecipitation assays indicated that PXR activation sequestrates the SRC‐1 coactivator from the HNF4α response element and E‐box of the OCT1 promoter. Consistent with these findings, exogenous overexpression of the SRC‐1, but not the PGC1α coactivator, relieved the PXR‐mediated repression of OCT1 transactivation. Conclusions and Implications PXR ligands reduced the HNF4α‐mediated and USF‐mediated transactivation of OCT1 gene expression by competing for SRC‐1 and decreased delivery of a model OCT1 substrate into hepatocytes. PMID:26920453
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Leach, Richard E; Kilburn, Brian A; Petkova, Anelia; Romero, Roberto; Armant, D Randall
2008-04-01
The antiapoptotic action of heparin-binding epidermal growth factor (HBEGF)-like growth factor and its regulation by O(2) constitutes a key factor for trophoblast survival. The hypothesis that cytotrophoblast survival is compromised by exposure to hypoxia-reoxygenation (H/R) injury, which may contribute to preeclampsia and some missed abortions, prompted us to investigate HBEGF regulation and its role as a survival factor during H/R in cytotrophoblast cells. A transformed human first-trimester cytotrophoblast cell line HTR-8/SVneo was exposed to H/R (2% O(2) followed by 20% O(2)) and assessed for HBEGF expression and cell death. Cellular HBEGF declined significantly within 30 minutes of reoxygenation after culture at 2% O(2). H/R significantly reduced proliferation and increased cell death when compared with trophoblast cells cultured continuously at 2% or 20% O(2). Restoration of cell survival also was achieved by adding recombinant HBEGF during reoxygenation. HBEGF inhibited apoptosis through its binding to either human epidermal receptor (HER)-1 or HER4, its cognate receptors. These results provide evidence that cytotrophoblast exposure to H/R induces apoptosis and decreased cell proliferation. HBEGF accumulation is diminished under these conditions, whereas restoration of HBEGF signaling improves trophoblast survival.
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Gupta, Vinayak; Khan, Abrar A; Sasi, Binu K; Mahapatra, Nitish R
2015-07-01
Monoamine oxidase A (MAOA) plays important roles in the pathogenesis of several neurological and cardiovascular disorders. The mechanism of transcriptional regulation of MAOA under basal and pathological conditions, however, remains incompletely understood. Here, we report systematic identification and characterization of cis elements and transcription factors that govern the expression of MAOA gene. Extensive computational analysis of MAOA promoter, followed by 5'-promoter deletion/reporter assays, revealed that the -71/-40 bp domain was sufficient for its basal transcription. Gel-shift and chromatin immunoprecipitation assays provided evidence of interactions of the transcription factors GATA-binding protein 2 (GATA2), Sp1 and TATA-binding protein (TBP) with this proximal promoter region. Consistently, over-expression of GATA2, Sp1 and TBP augmented MAOA promoter activity in a coordinated manner. In corroboration, siRNA-mediated down-regulation of GATA2/Sp1/TBP repressed the endogenous MAOA expression as well as transfected MAOA promoter activity. Tumor necrosis factor-α and forskolin activated MAOA transcription that was reversed by Sp1 siRNA; in support, tumor necrosis factor-α- and forskolin-induced activities were enhanced by ectopic over-expression of Sp1. On the other hand, MAOA transcription was diminished upon exposure of neuroblasts or cardiac myoblasts to ischemia-like conditions because of reduced binding of GATA2/Sp1/TBP with MAOA promoter. In conclusion, this study revealed previously unknown roles of GATA2, Sp1 and TBP in modulating MAOA expression under basal as well as pathophysiological conditions such as inflammation and ischemia, thus providing new insights into the molecular basis of aberrant MAOA expression in neuronal/cardiovascular disease states. Dysregulation of monoamine oxidase A (MAOA) have been implicated in several behavioral and neuronal disease states. Here, we identified three crucial transcription factors (GATA2, Sp1 and TBP) that regulate MAOA gene expression in a coordinated manner. Aberrant MAOA expression under pathophysiological conditions including inflammation and ischemia is mediated by altered binding of GATA2/Sp1/TBP with MAOA proximal promoter. Thus, these findings provide new insights into pathogenesis of several common diseases. GATA2, GATA-binding protein 2; Sp1, specificity protein 1; TBP, TATA-binding protein. © 2015 International Society for Neurochemistry.
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Bean Metal-Responsive Element-Binding Transcription Factor Confers Cadmium Resistance in Tobacco1
Sun, Na; Liu, Meng; Zhang, Wentao; Yang, Wanning; Bei, Xiujuan; Ma, Hui; Qiao, Fan; Qi, Xiaoting
2015-01-01
Cadmium (Cd) is highly toxic to plants. Modulation of Cd-responsive transcription is an important way for Cd detoxification in plants. Metal-responsive element (MRE) is originally described in animal metallothionein genes. Although functional MREs also exist in Cd-regulated plant genes, specific transcription factors that bind MRE to regulate Cd tolerance have not been identified. Previously, we showed that Cd-inducible bean (Phaseolus vulgaris) stress-related gene2 (PvSR2) produces a short (S) PvSR2 transcript (S-PvSR2) driven by an intronic promoter. Here, we demonstrate that S-PvSR2 encodes a bean MRE-binding transcription factor1 (PvMTF-1) that confers Cd tolerance in tobacco (Nicotiana tabacum). PvMTF-1 expression was up-regulated by Cd at the levels of RNA and protein. Importantly, expression of PvMTF-1 in tobacco enhanced Cd tolerance, indicating its role in regulating Cd resistance in planta. This was achieved through direct regulation of a feedback-insensitive Anthranilate Synthase α-2 chain gene (ASA2), which catalyzes the first step for tryptophan biosynthesis. In vitro and in vivo DNA-protein interaction studies further revealed that PvMTF-1 directly binds to the MRE in the ASA2 promoter, and this binding depends on the zinc finger-like motif of PvMTF-1. Through modulating ASA2 up-regulation by Cd, PvMTF-1 increased free tryptophan level and subsequently reduced Cd accumulation, thereby enhancing Cd tolerance of transgenic tobacco plants. Consistent with this observation, tobacco transiently overexpressing ASA2 also exhibited increased tolerance to Cd. We conclude that PvMTF-1 is a zinc finger-like transcription factor that links MRE to Cd resistance in transgenic tobacco through activation of tryptophan biosynthesis. PMID:25624396
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Brown, Sharron A N; Richards, Christine M; Hanscom, Heather N; Feng, Sheau-Line Y; Winkles, Jeffrey A
2003-01-01
Fn14 is a growth-factor-inducible immediate-early-response gene encoding a 102-amino-acid type I transmembrane protein. The human Fn14 protein was recently identified as a cell-surface receptor for the tumour necrosis factor (TNF) superfamily member named TWEAK (TNF-like weak inducer of apoptosis). In the present paper, we report that the human TWEAK extracellular domain can also bind the murine Fn14 protein. Furthermore, site-specific mutagenesis and directed yeast two-hybrid interaction assays revealed that the TNFR-associated factor (TRAF) 1, 2, 3 and 5 adaptor molecules bind the murine Fn14 cytoplasmic tail at an overlapping, but non-identical, amino acid sequence motif. We also found that TWEAK treatment of quiescent NIH 3T3 cells stimulates inhibitory kappaBalpha phosphorylation and transcriptional activation of a nuclear factor-kappaB (NF-kappaB) enhancer/luciferase reporter construct. Fn14 overexpression in transiently transfected NIH 3T3 cells also promotes NF-kappaB activation, and this cellular response requires an intact TRAF binding site. These results indicate that Fn14 is a functional TWEAK receptor that can associate with four distinct TRAF family members and stimulate the NF-kappaB transcription factor signalling pathway. PMID:12529173
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Chen, Xia; Liu, Liu; Chen, Yong; Yang, Yuting; Yang, Chao-Yie; Guo, Tianyue; Lei, Ming; Sun, Haiying; Wang, Shaomeng
2018-05-10
Telomeric repeat binding factor 2 (TRF2) is a telomere-associated protein that plays an important role in the formation of the 3' single strand DNA overhang and the "T loop", two structures critical for the stability of the telomeres. Apollo is a 5'-exonuclease recruited by TRF2 to the telomere and contributes to the formation of the 3' single strand DNA overhang. Knocking down of Apollo can induce DNA damage response similar to that caused by the knocking down of TRF2. In this Letter, we report the design and synthesis of a class of cyclic peptidic mimetics of the TRFH binding motif of Apollo (Apollo TBM ). We found conformational control of the C terminal residues of Apollo TBM can effectively improve the binding affinity. We have obtained a crystal structure of a cyclic peptidic Apollo peptide mimetic ( 34 ) complexed with TRF2, which provides valuable guidance to the future design of TRF2 inhibitors.
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Li, Guangyao; Zhou, Lei
2013-01-01
Due to the self-propagating nature of the heterochromatic modification H3K27me3, chromatin barrier activities are required to demarcate the boundary and prevent it from encroaching into euchromatic regions. Studies in Drosophila and vertebrate systems have revealed several important chromatin barrier elements and their respective binding factors. However, epigenomic data indicate that the binding of these factors are not exclusive to chromatin boundaries. To gain a comprehensive understanding of facultative heterochromatin boundaries, we developed a two-tiered method to identify the Chromatin Transitional Region (CTR), i.e. the nucleosomal region that shows the greatest transition rate of the H3K27me3 modification as revealed by ChIP-Seq. This approach was applied to identify CTRs in Drosophila S2 cells and human HeLa cells. Although many insulator proteins have been characterized in Drosophila, less than half of the CTRs in S2 cells are associated with known insulator proteins, indicating unknown mechanisms remain to be characterized. Our analysis also revealed that the peak binding of insulator proteins are usually 1–2 nucleosomes away from the CTR. Comparison of CTR-associated insulator protein binding sites vs. those in heterochromatic region revealed that boundary-associated binding sites are distinctively flanked by nucleosome destabilizing sequences, which correlates with significant decreased nucleosome density and increased binding intensities of co-factors. Interestingly, several subgroups of boundaries have enhanced H3.3 incorporation but reduced nucleosome turnover rate. Our genome-wide study reveals that diverse mechanisms are employed to define the boundaries of facultative heterochromatin. In both Drosophila and mammalian systems, only a small fraction of insulator protein binding sites co-localize with H3K27me3 boundaries. However, boundary-associated insulator binding sites are distinctively flanked by nucleosome destabilizing sequences, which correlates with significantly decreased nucleosome density and increased binding of co-factors. PMID:23840609
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Lead inhibition of DNA-binding mechanism of Cys(2)His(2) zinc finger proteins.
Hanas, J S; Rodgers, J S; Bantle, J A; Cheng, Y G
1999-11-01
The association of lead with chromatin in cells suggests that deleterious metal effects may in part be mediated through alterations in gene function. To elucidate if and how lead may alter DNA binding of cysteine-rich zinc finger proteins, lead ions were analyzed for their ability to alter the DNA binding mechanism of the Cys(2)His(2) zinc finger protein transcription factor IIIA (TFIIIA). As assayed by DNase I protection, the interaction of TFIIIA with the 50-bp internal control region of the 5S ribosomal gene was partially inhibited by 5 microM lead ions and completely inhibited by 10 to 20 microM lead ions. Preincubation of free TFIIIA with lead resulted in DNA-binding inhibition, whereas preincubation of a TFIIIA/5S RNA complex with lead did not result in DNA-binding inhibition. Because 5S RNA binds TFIIIA zinc fingers, this result is consistent with an inhibition mechanism via lead binding to zinc fingers. The complete loss of DNase I protection on the 5S gene indicates the mechanism of inhibition minimally involves the N-terminal fingers of TFIIIA. Inhibition was not readily reversible and occurred in the presence of an excess of beta-mercaptoethanol. Inhibition kinetics were fast, progressing to completion in approximately 5 min. Millimolar concentrations of sulfhydryl-specific arsenic ions were not inhibitory for TFIIIA binding. Micromolar concentrations of lead inhibited DNA binding by Sp1, another Cys(2)His(2) finger protein, but not by the nonfinger protein AP2. Inhibition of Cys(2)His(2) zinc finger transcription factors by lead ions at concentrations near those known to have deleterious physiological effects points to new molecular mechanisms for lead toxicity in promoting disease.
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Duque, Andrezza Marques; Leal, Márcia Carrera Campos; Marques, Ana Paula de Oliveira; Eskinazi, Fernanda Maria Vieira; Duque, Amanda Marques
2012-08-01
The scope of this paper was to determine the prevalence and factors associated with domestic violence against the elderly. It is a cross-sectional study with 274 subjects, aged 60 years or more, of both sexes. Data were collected through interviews at home or in the USF based on a script structured in three parts: questionnaire with socio-demographic and bio-demographic information, two rating scales and a tool for identifying abuse. Among the respondents, 20.8% reported having experienced at least one type of violence in their home environment. An association was revealed between those living with a greater number of individuals, among women and elderly people who are dependent for day-to-day activities. After applying the logistic regression model, only the variables of sex and family configuration were significantly associated, with evidence of greater frequency among those who lived with six or more residents and women. These findings highlight the magnitude and seriousness of the problem and point to the need for action to combat violence against the elderly.
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NASA Astrophysics Data System (ADS)
Luo, Benyi; Lu, Yigang
2008-10-01
Based on several hypotheses about the process of supercritical carbon dioxide extraction, the onflow around the solute granule is figured out by the Navier-Stocks equation. In combination with the Higbie’s solute infiltration model, the link between the mass-transfer coefficient and the velocity of flow is found. The mass-transfer coefficient with the ultrasonical effect is compared with that without the ultrasonical effect, and then a new parameter named the ultrasonic-enhanced factor of mass-transfer coefficient is brought forward, which describes the mathematical model of the supercritical carbon dioxide extraction process enhanced by ultrasonic. The model gives out the relationships among the ultrasonical power, the ultrasonical frequency, the radius of solute granule and the ultrasonic-enhanced factor of mass-transfer coefficient. The results calculated by this model fit well with the experimental data, including the extraction of Coix Lacryma-jobi Seed Oil (CLSO) and Coix Lacryma-jobi Seed Ester (CLSE) from coix seeds and the extraction of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) from the alga by means of the ultrasonic-enhanced supercritical carbon dioxide extraction (USFE) and the supercritical carbon dioxide extraction (SFE) respectively. This proves the rationality of the ultrasonic-enhanced factor model. The model provides a theoretical basis for the application of ultrasonic-enhanced supercritical fluid extraction technique.
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Genetic and epigenetic mechanisms of gene regulation during lens development
Cvekl, Ales; Duncan, Melinda K.
2007-01-01
Recent studies demonstrated a number of links between chromatin structure, gene expression, extracellular signaling and cellular differentiation during lens development. Lens progenitor cells originate from a pool of common progenitor cells, the pre-placodal region (PPR) which is formed due to a complex exchange of extracellular signals between the neural plate, naïve ectoderm and mesendoderm. A specific commitment to the lens program over alternate choices such as the formation of olfactory epithelium or the anterior pituitary is manifested by the formation of a thickened surface ectoderm, the lens placode. Mouse lens progenitor cells are characterized by the expression of a complement of lens lineage-specific transcription factors including Pax6, Six3 and Sox2, controlled by FGF and BMP signaling, followed later by c-Maf, Mab21like1, Prox1 and FoxE3. Proliferation of lens progenitors together with their morphogenetic movements results in the formation of the lens vesicle. This transient structure, comprised of lens precursor cells, is polarized with its anterior cells retaining their epithelial morphology and proliferative capacity, whereas the posterior lens precursor cells initiate terminal differentiation forming the primary lens fibers. Lens differentiation is marked by expression and accumulation of crystallins and other structural proteins. The transcriptional control of crystallin genes is characterized by the reiterative use of transcription factors required for the establishment of lens precursors in combination with more ubiquitously expressed factors (e.g. AP-1, AP-2α, CREB and USF) and recruitment of histone acetyltransferases (HATs) CBP and p300, and chromatin remodeling complexes SWI/SNF and ISWI. These studies have poised the study of lens development at the forefront of efforts to understand the connections between development, cell signaling, gene transcription and chromatin remodeling. PMID:17905638
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Kornhuber, J.; Mack-Burkhardt, F.; Konradi, C.
1989-01-01
The effect of a number of antemortem and postmortem factors on ({sup 3}H)MK-801 binding was investigated under equilibrium conditions in the frontal cortex of human brains of 38 controls. Binding values transiently increased during the early postnatal period reaching a maximum at the age of about 2 years. After age 10 years ({sup 3}H)MK-801 binding sites disappeared at 5.7% per decade. The storage time of brain tissue had a reducing effect on these binding sites. There was no effect of gender, brain weight or postmortem time interval and the binding sites were bilaterally symmetrically distributed in the frontal cortex.
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Gupta, Namrata; Gupta, Ankush; Kumar, Santosh; Mishra, Rajeev; Singh, Chhaya; Tripathi, Anil Kumar
2014-01-01
Azospirillum brasilense harbors two redox-sensitive Zinc-binding anti-sigma (ZAS) factors (ChrR1 and ChrR2), which negatively regulate the activity of their cognate extra-cytoplasmic function (ECF) σ factors (RpoE1 and RpoE2) by occluding their binding to the core enzyme. Both pairs of RpoE-ChrR control responses to photooxidative stress. The aim of this study was to investigate whether the two RpoE-ChrR pairs cross-talk while responding to the stress. In silico analysis showed a high sequence similarity between ChrR1 and ChrR2 proteins, but differences in redox sensitivity. Using in silico and in vitro methods of protein-protein interaction, we have shown that both ChrR1 and ChrR2 proteins physically bind to their noncognate RpoE proteins. Restoration of the phenotypes of chrR1::Tn5 and chrR2::Km mutants related to carotenoid biosynthesis and photooxidative stress tolerance by expressing chrR1 or chrR2 provided in vivo evidence for the cross-talk. In addition, up- or down-regulation of several identical proteins by expressing chrR1 or chrR2 in the chrR1::Tn5 mutant provided another in vivo evidence for the cross-talk. Although multiple redox-sensitive ZAS anti-σ factors occur in some Gram-positive bacteria, no cross-talk is reported among them. We report here, for the first time, that the two ZAS anti-σ factors of A. brasilense also interact with their noncognate σ factors and affect gene expression. The two redox-sensitive ZAS anti-σ factors in A. brasilense may interact with their cognate as well as noncognate ECF σ factors to play an important role in redox homeostasis by facilitating recovery from the oxidative stress.
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Monica S. Tomosy; Frank R. Thompson; Douglas Boyce
2011-01-01
This fall, the U.S. Forest Service (USFS) will release a comprehensive new guidebook designed to help managers develop climate adaptation options for National Forests (Peterson et al. 2011, in press). The adaptation process is based on partnerships between local resource managers and scientists working collaboratively to understand potential climate change effects,...
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6. View of HiattStricklin House showing north gable back and ...
6. View of Hiatt-Stricklin House showing north gable back and east side. Note the porches, shsutters and chimney. The shiny squares on the siding are metal pieces to repair woodpecker holes, facing southwest. - Hiatt Property, House, West bank of Woof Creek, 400 feet northwest of intersection of U.S.F.S. Roads 651 & 349, Placerville, Boise County, ID
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The soil indicator of forest health in the Forest Inventory and Analysis Program
Michael C. Amacher; Charles H. Perry
2010-01-01
Montreal Process Criteria and Indicators (MPCI) were established to monitor forest conditions and trends to promote sustainable forest management. The Soil Indicator of forest health was developed and implemented within the USFS Forest Inventory and Analysis (FIA) program to assess condition and trends in forest soil quality in U.S. forests regardless of ownership. The...
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DOT National Transportation Integrated Search
2007-10-15
At the request of the U.S. Department of Agriculture Forest Service (USFS), an inter-agency Transportation Assistance Group (TAG) site review of the status of planning and the options for providing alternative transportation at the El Yunque National...
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M. Matonis; R. Hubbard; K. Gebert; B. Hahn; C. Regan
2014-01-01
The Future Forest Webinar Series facilitated dialogue between scientists and managers about the challenges and opportunities created by the mountain pine beetle (MPB) epidemic. The series consisted of six webinar facilitated by the USFS Rocky Mountain Research Station, the Northern and Rocky Mountain Regions, and the Colorado Forest Restoration Institute. The series...
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Beach and Morphology Change Using Lidar
2016-11-01
Kelly R. Legault PURPOSE: This Coastal and Hydraulics Engineering Technical Note (CHETN) describes the use of lidar data in conjunction with beach...of Expertise. Beach profile surveys were provided by USACE Jacksonville District (SAJ), University of South Florida (USF), and Coastal Planning...within the limits of this study region include the Pinellas County Shoreline Protection Project (SPP) (USACE SAJ 2010), Tampa Harbor Navigation
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Rachel Riemann; Barry Tyler Wilson; Andrew Lister; Sarah Parks
2010-01-01
Geospatial datasets of forest characteristics are modeled representations of real populations on the ground. The continuous spatial character of such datasets provides an incredible source of information at the landscape level for ecosystem research, policy analysis, and planning applications, all of which are critical for addressing current challenges related to...
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Fort Valley Experimental Forest-A Century of Research 1908-2008
Susan D. Olberding; Margaret M. Moore
2008-01-01
One hundred years ago, the USFS began its forest research program in a two-room cabin near Flagstaff, Arizona, with one staff person, Gustaf A. Pearson. The site became known as the Fort Valley Experiment Station and was the first in a national network of research sites developed to address uncertainties regarding the rehabilitation and conservation for forest and...
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Fort Valley Experimental Forest-A Century of Research 1908-2008 (P-53)
Susan D. Olberding; Margaret M. Moore
2008-01-01
One hundred years ago, the USFS began its forest research program in a two-room cabin near Flagstaff, Arizona, with one staff person, Gustaf A. Pearson. The site became known as the Fort Valley Experiment Station and was the first in a national network of research sites developed to address uncertainties regarding the rehabilitation and conservation for forest and...
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Effects of climate change on ecosystem services in the Northern Rockies Region [Chapter 11
Travis Warziniack; Megan Lawson; S. Karen Dante-Wood
2018-01-01
In this chapter, we focus on the ecosystem services provided to people who visit, live adjacent to, or otherwise benefit from natural resources on public lands. Communities in the Forest Service, U.S. Department of Agriculture (USFS) Northern Region and the Greater Yellowstone Area (GYA), hereafter called the Northern Rockies region, are highly dependent on ecosystem...
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The North Fork of Caspar Creek: a cooperative venture between CDF and USFS
Pete Cafferata
1984-01-01
The Caspar Creek Watershed Study on JDSF has taken a new direction in the last two years, as our work progresses towards full instrumentation of the North Fork phase. When most of the equipment has been installed by the end of the summer, this 1195-acre watershed will become the most intensively sampled drainage ever studied by hydrologists.
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Releases of natural enemies in Hawaii since 1980 for classical biological control of weeds
P. Conant; J. N. Garcia; M. T. Johnson; W. T. Nagamine; C. K. Hirayama; G. P. Markin; R. L. Hill
2013-01-01
A comprehensive review of biological control of weeds in Hawaii was last published in 1992, covering 74 natural enemy species released from 1902 through 1980. The present review summarizes releases of 21 natural enemies targeting seven invasive weeds from 1981 to 2010. These projects were carried out by Hawaii Department of Agriculture (HDOA), USDA Forest Service (USFS...
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Pennsylvania's experiences with microbial control of the gypsy moth
James O. Nichols
1985-01-01
Pennsylvania's first experience with using Bt on insect control occurred in 1964. For the next 17 years, various projects were conducted, in cooperation with the USFS and industry, in an effort to secure operational status of Bt for gypsy moth suppression. This point was reached in 1982, and the Governor was convinced that the time was right to convert most gypsy...
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Designing economic impact assessments for USFS wildfire programs
Karen L. Abt; Robert J. Jr. Huggett; Thomas P. Holmes
2008-01-01
As often happens in the wake of a series of extreme fire seasons, such as those in 2000, 2002 and 2003, federal wildfire policy is being scrutinized and recommendations regarding changes both large and small are prevalent (Stephens and Ruth 2005, Busenberg 2004, Dellasalla et al. 2004, Dombeck et al. 2004). It is common practice for increases in acres burned and in...
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Susan D. Olberding; Karen Malis-Clark; Peter J. Pilles; Dennis Lund
2008-01-01
This poster presents the continuing cooperative relationship between the Fort Valley Experimental Forest (FVEF), Coconino National Forest (CNF), USFS Region 3, and the long-term partnerships with the Museum of the Northern Arizona and the NAU School of Forestry.Fort Valley was initially named the Coconino Experiment Station and funds were channeled...
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Susan D. Olberding; Karen Malis-Clark; Peter J. Pilles; Dennis Lund
2008-01-01
This poster presents the continuing cooperative relationship between the Fort Valley Experimental Forest (FVEF), Coconino National Forest (CNF), USFS Region 3, and the long-term partnerships with the Museum of the Northern Arizona and the NAU School of Forestry. Fort Valley was initially named the Coconino Experiment Station and funds were channeled...
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A Search for TRUTH in Student Responses to Selected Survey Items. AIR 1993 Annual Forum Paper.
ERIC Educational Resources Information Center
Takalkar, Pradnya; And Others
This study compared 4,594 student responses from three different surveys of incoming students at the University of South Florida (USF) with data from Florida's State University System (SUS) admissions files to determine what proportion of error occurs in the survey responses. Specifically, the study investigated the amount of measurement error in…
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ERIC Educational Resources Information Center
Universal Service Administrative Company, 2007
2007-01-01
This report shows how Universal Service Fund support for schools and libraries is used by school districts and libraries around the country. Highlighted are approximately 190 success stories of program participants that have come to rely on the USF to expand educational opportunities for students through better use of telecommunications technology…
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A tool to analyze environmental impacts of roads on forest watersheds
Ajay Prasad
2007-01-01
The construction and use of forest roads can have impacts on geomorphic processes and erosion patterns in forested basins. Analyzing these impacts will help forest managers to effectively manage road and road drainage system and hence minimize the negative impacts of forest roads. To manage forest roads effectively the USDA Forest Service (USFS) has developed a road...
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Sensitivity of Landsat image-derived burn severity indices to immediate post-fire effects
A. T. Hudak; S. Lewis; P. Robichaud; P. Morgan; M. Bobbitt; L. Lentile; A. Smith; Z. Holden; J. Clark; R. McKinley
2006-01-01
The USFS Remote Sensing Applications Center (RSAC) and the USGS Center for Earth Resources Observation and Science (EROS) produce Burned Area Reflectance Classification (BARC) maps as a rapid, preliminary indication of burn severity on large wildfire events. Currently the preferred burn severity index is the delta Normalized Burn Ratio (dNBR), which requires NBR values...
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Characterizing large airtanker use in United States fire management
Crystal S. Stonesifer; Matthew P. Thompson; Dave Calkin; Charles W. McHugh
2015-01-01
The appropriate role of large airtankers (LATs) in federal fire suppression in the United States has been the source of much debate and discussion in recent years as the U.S. Forest Service (USFS) has faced impending decisions about how best to address an aging fleet of contracted aircraft. Questions of fleet efficiency are complicated by inadequacies in historical...
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Historic and Contemporary Land Use in Southwestern Grassland Ecosystems
Carol Raish
2004-01-01
This chapter encompasses the lands of the Southwest as defined by Region 3 of the USDA Forest Service (USFS): Arizona, New Mexico, and portions of western Oklahoma and the Texas Panhandle. I examine human use and modification of the grasslands/rangelands of this region, with an emphasis on those areas managed by the Forest Service. Because the majority of publications...
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Forest Inventory and Analysis National Data Quality Assessment Report for 2000 to 2003
James E. Pollard; James A. Westfall; Paul L. Patterson; David L. Gartner; Mark Hansen; Olaf Kuegler
2006-01-01
The Forest Inventory and Analysis program (FIA) is the key USDA Forest Service (USFS) program that provides the information needed to assess the status and trends in the environmental quality of the Nation's forests. The goal of the FIA Quality Assurance (QA) program is to provide a framework to assure the production of complete, accurate and unbiased forest...
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U.S. Forest Service Region 1 Lake Chemistry, NADP, and IMPROVE air quality data analysis
Jill Grenon; Mark Story
2009-01-01
This report was developed to address the need for comprehensive analysis of U.S. Forest Service (USFS) Region 1 air quality monitoring data. The monitoring data includes Phase 3 (long-term data) lakes, National Atmospheric Deposition Program (NADP), and Interagency Monitoring of Protected Visual Environments (IMPROVE). Annual and seasonal data for the periods of record...
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Forest and range research on the "Wild Bill Plots" (1927-2007)
Daniel C. Laughlin; Margaret M. Moore
2008-01-01
In 1927, the Fort Valley Experimental Forest initiated a range-timber reproduction study. The study was one of the first attempts to experimentally isolate the agents responsible for injury to ponderosa pine regeneration, and at the same time assess the impacts of livestock grazing on herbaceous vegetation. The study was conducted on the USFS range allotments northwest...
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Forest and range research on the "Wild Bill Plots" (1927-2007) (P-53)
Daniel C. Laughlin; Margaret M. Moore
2008-01-01
In 1927, the Fort Valley Experimental Forest initiated a range-timber reproduction study. The study was one of the first attempts to experimentally isolate the agents responsible for injury to ponderosa pine regeneration, and at the same time assess the impacts of livestock grazing on herbaceous vegetation. The study was conducted on the USFS range allotments northwest...
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Interaction of AIM with insulin-like growth factor-binding protein-4
YOU, QIANG; WU, YAN; YAO, NANNAN; SHEN, GUANNAN; ZHANG, YING; XU, LIANGGUO; LI, GUIYING; JU, CYNTHIA
2015-01-01
Apoptosis inhibitor of macrophages (AIM/cluster of differentiation 5 antigen-like/soluble protein α) has been shown to inhibit cellular apoptosis; however, the underlying molecular mechanism has not been elucidated. Using yeast two-hybrid screening, the present study uncovered that AIM binds to insulin-like growth factor binding protein-4 (IGFBP-4). AIM interaction with IGFBP-4, as well as IGFBP-2 and -3, but not with IGFBP-1, -5 and -6, was further confirmed by co-immunoprecipitation (co-IP) using 293 cells. The binding activity and affinity between AIM and IGFBP-4 in vitro were analyzed by co-IP and biolayer interferometry. Serum depletion-induced cellular apoptosis was attenuated by insulin-like growth factor-I (IGF-I), and this effect was abrogated by IGFBP-4. Of note, in the presence of AIM, the inhibitory effect of IGFBP-4 on the anti-apoptosis function of IGF-I was attenuated, possibly through binding of AIM with IGFBP-4. In conclusion, to the best of our knowledge, the present study provides the first evidence that AIM binds to IGFBP-2, -3 and -4. The data suggest that this interaction may contribute to the mechanism of AIM-mediated anti-apoptosis function. PMID:26135353
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Wang, Guohua; Wang, Fang; Huang, Qian; Li, Yu; Liu, Yunlong; Wang, Yadong
2015-01-01
Transcription factors are proteins that bind to DNA sequences to regulate gene transcription. The transcription factor binding sites are short DNA sequences (5-20 bp long) specifically bound by one or more transcription factors. The identification of transcription factor binding sites and prediction of their function continue to be challenging problems in computational biology. In this study, by integrating the DNase I hypersensitive sites with known position weight matrices in the TRANSFAC database, the transcription factor binding sites in gene regulatory region are identified. Based on the global gene expression patterns in cervical cancer HeLaS3 cell and HelaS3-ifnα4h cell (interferon treatment on HeLaS3 cell for 4 hours), we present a model-based computational approach to predict a set of transcription factors that potentially cause such differential gene expression. Significantly, 6 out 10 predicted functional factors, including IRF, IRF-2, IRF-9, IRF-1 and IRF-3, ICSBP, belong to interferon regulatory factor family and upregulate the gene expression levels responding to the interferon treatment. Another factor, ISGF-3, is also a transcriptional activator induced by interferon alpha. Using the different transcription factor binding sites selected criteria, the prediction result of our model is consistent. Our model demonstrated the potential to computationally identify the functional transcription factors in gene regulation.
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Kowenz-Leutz, Elisabeth; Schuetz, Anja; Liu, Qingbin; Knoblich, Maria; Heinemann, Udo; Leutz, Achim
2016-07-01
The transcription factor CCAAT/enhancer-binding protein α (C/EBPα) regulates cell cycle arrest and terminal differentiation of neutrophils and adipocytes. Mutations in the basic leucine zipper domain (bZip) of C/EBPα are associated with acute myeloid leukemia. A widely used murine transforming C/EBPα basic region mutant (BRM2) entails two bZip point mutations (I294A/R297A). BRM2 has been discordantly described as defective for DNA binding or defective for interaction with E2F. We have separated the two BRM2 mutations to shed light on the intertwined reciprocity between C/EBPα-E2F-DNA interactions. Both, C/EBPα I294A and R297A retain transactivation capacity and interaction with E2F-DP. The C/EBPα R297A mutation destabilized DNA binding, whereas the C/EBPα I294A mutation enhanced binding to DNA. The C/EBPα R297A mutant, like BRM2, displayed enhanced interaction with E2F-DP but failed to repress E2F-dependent transactivation although both mutants were readily suppressed by E2F1 for transcription through C/EBP cis-regulatory sites. In contrast, the DNA binding enhanced C/EBPα I294A mutant displayed increased repression of E2F-DP mediated transactivation and resisted E2F-DP mediated repression. Thus, the efficient repression of E2F dependent S-phase genes and the activation of differentiation genes reside in the balanced DNA binding capacity of C/EBPα. Copyright © 2016 Elsevier B.V. All rights reserved.
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Weissbach, Julia; Schikora, Franziska; Weber, Anja; Kessels, Michael
2016-01-01
The myocardin-related transcription factors (MRTFs) are coactivators of serum response factor (SRF)-mediated gene expression. Activation of MRTF-A occurs in response to alterations in actin dynamics and critically requires the dissociation of repressive G-actin–MRTF-A complexes. However, the mechanism leading to the release of MRTF-A remains unclear. Here we show that WH2 domains compete directly with MRTF-A for actin binding. Actin nucleation-promoting factors, such as N-WASP and WAVE2, as well as isolated WH2 domains, including those of Spire2 and Cobl, activate MRTF-A independently of changes in actin dynamics. Simultaneous inhibition of Arp2-Arp3 or mutation of the CA region only partially reduces MRTF-A activation by N-WASP and WAVE2. Recombinant WH2 domains and the RPEL domain of MRTF-A bind mutually exclusively to cellular and purified G-actin in vitro. The competition by different WH2 domains correlates with MRTF-SRF activation. Following serum stimulation, nonpolymerizable actin dissociates from MRTF-A, and de novo formation of the G-actin–RPEL complex is impaired by a transferable factor. Our work demonstrates that WH2 domains activate MRTF-A and contribute to target gene regulation by a competitive mechanism, independently of their role in actin filament formation. PMID:26976641
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Fukutomi, Toshiaki; Takagi, Kenji; Mizushima, Tsunehiro; Ohuchi, Noriaki
2014-01-01
Transcription factor Nrf2 (NF-E2-related factor 2) coordinately regulates cytoprotective gene expression, but under unstressed conditions, Nrf2 is degraded rapidly through Keap1 (Kelch-like ECH-associated protein 1)-mediated ubiquitination. Nrf2 harbors two Keap1-binding motifs, DLG and ETGE. Interactions between these two motifs and Keap1 constitute a key regulatory nexus for cellular Nrf2 activity through the formation of a two-site binding hinge-and-latch mechanism. In this study, we determined the minimum Keap1-binding sequence of the DLG motif, the low-affinity latch site, and defined a new DLGex motif that covers a sequence much longer than that previously defined. We have successfully clarified the crystal structure of the Keap1-DC-DLGex complex at 1.6 Å. DLGex possesses a complicated helix structure, which interprets well the human-cancer-derived loss-of-function mutations in DLGex. In thermodynamic analyses, Keap1-DLGex binding is characterized as enthalpy and entropy driven, while Keap1-ETGE binding is characterized as purely enthalpy driven. In kinetic analyses, Keap1-DLGex binding follows a fast-association and fast-dissociation model, while Keap1-ETGE binding contains a slow-reaction step that leads to a stable conformation. These results demonstrate that the mode of DLGex binding to Keap1 is distinct from that of ETGE structurally, thermodynamically, and kinetically and support our contention that the DLGex motif serves as a converter transmitting environmental stress to Nrf2 induction as the latch site. PMID:24366543
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Kozlowski, Maya; Larose, Louise; Lee, Fai; Le, Duc Mingh; Rottapel, Robert; Siminovitch, Katherine A.
1998-01-01
The SH2 domain-containing SHP-1 tyrosine phosphatase has been shown to negatively regulate a broad spectrum of growth factor- and cytokine-driven mitogenic signaling pathways. Included among these is the cascade of intracellular events evoked by stem cell factor binding to c-Kit, a tyrosine kinase receptor which associates with and is dephosphorylated by SHP-1. Using a series of glutathione S-transferase (GST) fusion proteins containing either tyrosine-phosphorylated segments of the c-Kit cytosolic region or the SH2 domains of SHP-1, we have shown that SHP-1 interacts with c-Kit by binding selectively to the phosphorylated c-Kit juxtamembrane region and that the association of c-Kit with the larger of the two SHP-1 isoforms may be mediated through either the N-terminal or C-terminal SHP-1 SH2 domain. The results of binding assays with mutagenized GST-Kit juxtamembrane fusion proteins and competitive inhibition assays with phosphopeptides encompassing each c-Kit juxtamembrane region identified the tyrosine residue at position 569 as the major site for binding of SHP-1 to c-Kit and suggested that tyrosine 567 contributes to, but is not required for, this interaction. By analysis of Ba/F3 cells retrovirally transduced to express c-Kit receptors, phenylalanine substitution of c-Kit tyrosine residue 569 was shown to be associated with disruption of c-Kit–SHP-1 binding and induction of hyperproliferative responses to stem cell factor. Although phenylalanine substitution of c-Kit tyrosine residue 567 in the Ba/F3–c-Kit cells did not alter SHP-1 binding to c-Kit, the capacity of a second c-Kit-binding tyrosine phosphatase, SHP-2, to associate with c-Kit was markedly reduced, and the cells again showed hyperproliferative responses to stem cell factor. These data therefore identify SHP-1 binding to tyrosine 569 on c-Kit as an interaction pivotal to SHP-1 inhibitory effects on c-Kit signaling, but they indicate as well that cytosolic protein tyrosine phosphatases other than SHP-1 may also negatively regulate the coupling of c-Kit engagement to proliferation. PMID:9528781
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Narayan, Vikram; Halada, Petr; Hernychová, Lenka; Chong, Yuh Ping; Žáková, Jitka; Hupp, Ted R; Vojtesek, Borivoj; Ball, Kathryn L
2011-04-22
The interferon-regulated transcription factor and tumor suppressor protein IRF-1 is predicted to be largely disordered outside of the DNA-binding domain. One of the advantages of intrinsically disordered protein domains is thought to be their ability to take part in multiple, specific but low affinity protein interactions; however, relatively few IRF-1-interacting proteins have been described. The recent identification of a functional binding interface for the E3-ubiquitin ligase CHIP within the major disordered domain of IRF-1 led us to ask whether this region might be employed more widely by regulators of IRF-1 function. Here we describe the use of peptide aptamer-based affinity chromatography coupled with mass spectrometry to define a multiprotein binding interface on IRF-1 (Mf2 domain; amino acids 106-140) and to identify Mf2-binding proteins from A375 cells. Based on their function as known transcriptional regulators, a selection of the Mf2 domain-binding proteins (NPM1, TRIM28, and YB-1) have been validated using in vitro and cell-based assays. Interestingly, although NPM1, TRIM28, and YB-1 all bind to the Mf2 domain, they have differing amino acid specificities, demonstrating the degree of combinatorial diversity and specificity available through linear interaction motifs.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Bach, Christian; Sherman, William; Pallis, Jani
Zinc finger nucleases (ZFNs) are associated with cell death and apoptosis by binding at countless undesired locations. This cytotoxicity is associated with the binding ability of engineered zinc finger domains to bind dissimilar DNA sequences with high affinity. In general, binding preferences of transcription factors are associated with significant degenerated diversity and complexity which convolutes the design and engineering of precise DNA binding domains. Evolutionary success of natural zinc finger proteins, however, evinces that nature created specific evolutionary traits and strategies, such as modularity and rank-specific recognition to cope with binding complexity that are critical for creating clinical viable toolsmore » to precisely modify the human genome. Our findings indicate preservation of general modularity and significant alteration of the rank-specific binding preferences of the three-finger binding domain of transcription factor SP1 when exchanging amino acids in the 2nd finger.« less
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Foti, M; Omichinski, J G; Stahl, S; Maloney, D; West, J; Schweitzer, B I
1999-02-05
We investigate here the effects of the incorporation of the nucleoside analogs araC (1-beta-D-arabinofuranosylcytosine) and ganciclovir (9-[(1,3-dihydroxy-2-propoxy)methyl] guanine) into the DNA binding recognition sequence for the GATA-1 erythroid transcription factor. A 10-fold decrease in binding affinity was observed for the ganciclovir-substituted DNA complex in comparison to an unmodified DNA of the same sequence composition. AraC substitution did not result in any changes in binding affinity. 1H-15N HSQC and NOESY NMR experiments revealed a number of chemical shift changes in both DNA and protein in the ganciclovir-modified DNA-protein complex when compared to the unmodified DNA-protein complex. These changes in chemical shift and binding affinity suggest a change in the binding mode of the complex when ganciclovir is incorporated into the GATA DNA binding site.
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Bach, Christian; Sherman, William; Pallis, Jani; ...
2014-01-01
Zinc finger nucleases (ZFNs) are associated with cell death and apoptosis by binding at countless undesired locations. This cytotoxicity is associated with the binding ability of engineered zinc finger domains to bind dissimilar DNA sequences with high affinity. In general, binding preferences of transcription factors are associated with significant degenerated diversity and complexity which convolutes the design and engineering of precise DNA binding domains. Evolutionary success of natural zinc finger proteins, however, evinces that nature created specific evolutionary traits and strategies, such as modularity and rank-specific recognition to cope with binding complexity that are critical for creating clinical viable toolsmore » to precisely modify the human genome. Our findings indicate preservation of general modularity and significant alteration of the rank-specific binding preferences of the three-finger binding domain of transcription factor SP1 when exchanging amino acids in the 2nd finger.« less
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Miles, Wayne O; Dyson, Nicholas J
2014-01-01
The ability of the retinoblastoma protein (RB) tumor suppressor to repress transcription stimulated by the E2 promoter binding factors (E2F) is integral to its biological functions. Our recent report described a conserved feedback mechanism mediated by the RNA-binding proteins Pumilio and Nanos that increases in importance following RB loss and helps cells to tolerate deregulated E2F. PMID:27308363
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Miles, Wayne O; Dyson, Nicholas J
2014-01-01
The ability of the retinoblastoma protein (RB) tumor suppressor to repress transcription stimulated by the E2 promoter binding factors (E2F) is integral to its biological functions. Our recent report described a conserved feedback mechanism mediated by the RNA-binding proteins Pumilio and Nanos that increases in importance following RB loss and helps cells to tolerate deregulated E2F.
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Structural basis for genome wide recognition of 5-bp GC motifs by SMAD transcription factors.
Martin-Malpartida, Pau; Batet, Marta; Kaczmarska, Zuzanna; Freier, Regina; Gomes, Tiago; Aragón, Eric; Zou, Yilong; Wang, Qiong; Xi, Qiaoran; Ruiz, Lidia; Vea, Angela; Márquez, José A; Massagué, Joan; Macias, Maria J
2017-12-12
Smad transcription factors activated by TGF-β or by BMP receptors form trimeric complexes with Smad4 to target specific genes for cell fate regulation. The CAGAC motif has been considered as the main binding element for Smad2/3/4, whereas Smad1/5/8 have been thought to preferentially bind GC-rich elements. However, chromatin immunoprecipitation analysis in embryonic stem cells showed extensive binding of Smad2/3/4 to GC-rich cis-regulatory elements. Here, we present the structural basis for specific binding of Smad3 and Smad4 to GC-rich motifs in the goosecoid promoter, a nodal-regulated differentiation gene. The structures revealed a 5-bp consensus sequence GGC(GC)|(CG) as the binding site for both TGF-β and BMP-activated Smads and for Smad4. These 5GC motifs are highly represented as clusters in Smad-bound regions genome-wide. Our results provide a basis for understanding the functional adaptability of Smads in different cellular contexts, and their dependence on lineage-determining transcription factors to target specific genes in TGF-β and BMP pathways.
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DNA-binding regulates site-specific ubiquitination of IRF-1.
Landré, Vivien; Pion, Emmanuelle; Narayan, Vikram; Xirodimas, Dimitris P; Ball, Kathryn L
2013-02-01
Understanding the determinants for site-specific ubiquitination by E3 ligase components of the ubiquitin machinery is proving to be a challenge. In the present study we investigate the role of an E3 ligase docking site (Mf2 domain) in an intrinsically disordered domain of IRF-1 [IFN (interferon) regulatory factor-1], a short-lived IFNγ-regulated transcription factor, in ubiquitination of the protein. Ubiquitin modification of full-length IRF-1 by E3 ligases such as CHIP [C-terminus of the Hsc (heat-shock cognate) 70-interacting protein] and MDM2 (murine double minute 2), which dock to the Mf2 domain, was specific for lysine residues found predominantly in loop structures that extend from the DNA-binding domain, whereas no modification was detected in the more conformationally flexible C-terminal half of the protein. The E3 docking site was not available when IRF-1 was in its DNA-bound conformation and cognate DNA-binding sequences strongly suppressed ubiquitination, highlighting a strict relationship between ligase binding and site-specific modification at residues in the DNA-binding domain. Hyperubiquitination of a non-DNA-binding mutant supports a mechanism where an active DNA-bound pool of IRF-1 is protected from polyubiquitination and degradation.
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TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains
Ollagnier, Guillaume; Désiré, Nathalie; Sayon, Sophie; Raingeaud, Jöel; Marcelin, Anne-Geneviève; Calvez, Vincent; Khammari, Amir; Batteux, Frédéric; Dréno, Brigitte; Dupin, Nicolas
2016-01-01
Background Propionibacterium acnes (P. acnes) is an anaerobic, Gram-positive bacteria encountered in inflammatory acne lesions, particularly in the pilosebaceous follicle. P. acnes triggers a strong immune response involving keratinocytes, sebocytes and monocytes, the target cells during acne development. Lipoteicoic acid and peptidoglycan induce the inflammatory reaction, but no P. acnes surface protein interacting with Toll-like receptors has been identified. P. acnes surface proteins have been extracted by lithium stripping and shown to induce CXCL8 production by keratinocytes. Methodology and principal findings Far-western blotting identified two surface proteins, of 24.5- and 27.5-kDa in size, specifically recognized by TLR2. These proteins were characterized, by LC-MS/MS, as CAMP factor 1 devoid of its signal peptide sequence, as shown by N-terminal sequencing. Purified CAMP factor 1 induces CXCL8 production by activating the CXCL8 gene promoter, triggering the synthesis of CXCL8 mRNA. Antibodies against TLR2 significantly decreased the CXCL8 response. For the 27 P. acnes strains used in this study, CAMP1-TLR2 binding intensity was modulated and appeared to be strong in type IB and II strains, which produced large amounts of CXCL8, whereas most of the type IA1 and IA2 strains presented little or no CAMP1-TLR2 binding and low levels of CXCL8 production. The nucleotide sequence of CAMP factor displays a major polymorphism, defining two distinct genetic groups corresponding to CAMP factor 1 with 14 amino-acid changes from strains phylotyped II with moderate and high levels of CAMP1-TLR2 binding activity, and CAMP factor 1 containing 0, 1 or 2 amino-acid changes from strains phylotyped IA1, IA2, or IB presenting no, weak or moderate CAMP1-TLR2 binding. Conclusions Our findings indicate that CAMP factor 1 may contribute to P. acnes virulence, by amplifying the inflammation reaction through direct interaction with TLR2. PMID:27902761
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TLR-2 Recognizes Propionibacterium acnes CAMP Factor 1 from Highly Inflammatory Strains.
Lheure, Coralie; Grange, Philippe Alain; Ollagnier, Guillaume; Morand, Philippe; Désiré, Nathalie; Sayon, Sophie; Corvec, Stéphane; Raingeaud, Jöel; Marcelin, Anne-Geneviève; Calvez, Vincent; Khammari, Amir; Batteux, Frédéric; Dréno, Brigitte; Dupin, Nicolas
2016-01-01
Propionibacterium acnes (P. acnes) is an anaerobic, Gram-positive bacteria encountered in inflammatory acne lesions, particularly in the pilosebaceous follicle. P. acnes triggers a strong immune response involving keratinocytes, sebocytes and monocytes, the target cells during acne development. Lipoteicoic acid and peptidoglycan induce the inflammatory reaction, but no P. acnes surface protein interacting with Toll-like receptors has been identified. P. acnes surface proteins have been extracted by lithium stripping and shown to induce CXCL8 production by keratinocytes. Far-western blotting identified two surface proteins, of 24.5- and 27.5-kDa in size, specifically recognized by TLR2. These proteins were characterized, by LC-MS/MS, as CAMP factor 1 devoid of its signal peptide sequence, as shown by N-terminal sequencing. Purified CAMP factor 1 induces CXCL8 production by activating the CXCL8 gene promoter, triggering the synthesis of CXCL8 mRNA. Antibodies against TLR2 significantly decreased the CXCL8 response. For the 27 P. acnes strains used in this study, CAMP1-TLR2 binding intensity was modulated and appeared to be strong in type IB and II strains, which produced large amounts of CXCL8, whereas most of the type IA1 and IA2 strains presented little or no CAMP1-TLR2 binding and low levels of CXCL8 production. The nucleotide sequence of CAMP factor displays a major polymorphism, defining two distinct genetic groups corresponding to CAMP factor 1 with 14 amino-acid changes from strains phylotyped II with moderate and high levels of CAMP1-TLR2 binding activity, and CAMP factor 1 containing 0, 1 or 2 amino-acid changes from strains phylotyped IA1, IA2, or IB presenting no, weak or moderate CAMP1-TLR2 binding. Our findings indicate that CAMP factor 1 may contribute to P. acnes virulence, by amplifying the inflammation reaction through direct interaction with TLR2.
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OsDREB2A, a Rice Transcription Factor, Significantly Affects Salt Tolerance in Transgenic Soybean
Ma, Qi-bin; Yang, Cun-yi; Mu, Ying-hui; Suo, Hai-cui; Luo, Lai-hui; Nian, Hai
2013-01-01
The dehydration responsive element binding (DREB) transcription factors play an important role in regulating stress-related genes. OsDREB2A, a member of the DREBP subfamily of AP2/ERF transcription factors in rice (Oryza sativa), is involved in the abiotic stress response. OsDREB2A expression is induced by drought, low-temperature and salt stresses. Here, we report the ability of OsDREB2A to regulate high-salt response in transgenic soybean. Overexpressing OsDREB2A in soybeans enhanced salt tolerance by accumulating osmolytes, such as soluble sugars and free proline, and improving the expression levels of some stress-responsive transcription factors and key genes. The phenotypic characterization of transgenic soybean were significantly better than those of wild-type (WT). Electrophoresis mobility shift assay (EMSA) revealed that the OsDREB2A can bind to the DRE core element in vitro. These results indicate that OsDREB2A may participate in abiotic stress by directly binding with DRE element to regulate the expression of downstream genes. Overexpression of OsDREB2A in soybean might be used to improve tolerance to salt stress. PMID:24376625
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OnTheFly: a database of Drosophila melanogaster transcription factors and their binding sites.
Shazman, Shula; Lee, Hunjoong; Socol, Yakov; Mann, Richard S; Honig, Barry
2014-01-01
We present OnTheFly (http://bhapp.c2b2.columbia.edu/OnTheFly/index.php), a database comprising a systematic collection of transcription factors (TFs) of Drosophila melanogaster and their DNA-binding sites. TFs predicted in the Drosophila melanogaster genome are annotated and classified and their structures, obtained via experiment or homology models, are provided. All known preferred TF DNA-binding sites obtained from the B1H, DNase I and SELEX methodologies are presented. DNA shape parameters predicted for these sites are obtained from a high throughput server or from crystal structures of protein-DNA complexes where available. An important feature of the database is that all DNA-binding domains and their binding sites are fully annotated in a eukaryote using structural criteria and evolutionary homology. OnTheFly thus provides a comprehensive view of TFs and their binding sites that will be a valuable resource for deciphering non-coding regulatory DNA.
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2018-01-01
ABSTRACT Herpes simplex virus 1 (HSV-1) establishes latent infection in neurons via a variety of epigenetic mechanisms that silence its genome. The cellular CCCTC-binding factor (CTCF) functions as a mediator of transcriptional control and chromatin organization and has binding sites in the HSV-1 genome. We constructed an HSV-1 deletion mutant that lacked a pair of CTCF-binding sites (CTRL2) within the latency-associated transcript (LAT) coding sequences and found that loss of these CTCF-binding sites did not alter lytic replication or levels of establishment of latent infection, but their deletion reduced the ability of the virus to reactivate from latent infection. We also observed increased heterochromatin modifications on viral chromatin over the LAT promoter and intron. We therefore propose that CTCF binding at the CTRL2 sites acts as a chromatin insulator to keep viral chromatin in a form that is poised for reactivation, a state which we call poised latency. PMID:29437926
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The Evolving Field of Human Papillomavirus Receptor Research: a Review of Binding and Entry
Raff, Adam B.; Woodham, Andrew W.; Raff, Laura M.; Skeate, Joseph G.; Yan, Lisa; Da Silva, Diane M.; Schelhaas, Mario
2013-01-01
Human papillomaviruses (HPVs) infect epithelia and can lead to the development of lesions, some of which have malignant potential. HPV type 16 (HPV16) is the most oncogenic genotype and causes various types of cancer, including cervical, anal, and head and neck cancers. However, despite significant research, our understanding of the mechanism by which HPV16 binds to and enters host cells remains fragmented. Over several decades, many HPV receptors and entry pathways have been described. This review puts those studies into context and offers a model of HPV16 binding and entry as a framework for future research. Our model suggests that HPV16 binds to heparin sulfate proteoglycans (HSPGs) on either the epithelial cell surface or basement membrane through interactions with the L1 major capsid protein. Growth factor receptors may also become activated through HSPG/growth factor/HPV16 complexes that initiate signaling cascades during early virion-host cell interactions. After binding to HSPGs, the virion undergoes conformational changes, leading to isomerization by cyclophilin B and proprotein convertase-mediated L2 minor capsid protein cleavage that increases L2 N terminus exposure. Along with binding to HSPGs, HPV16 binds to α6 integrins, which initiate further intracellular signaling events. Following these primary binding events, HPV16 binds to a newly identified L2-specific receptor, the annexin A2 heterotetramer. Subsequently, clathrin-, caveolin-, lipid raft-, flotillin-, cholesterol-, and dynamin-independent endocytosis of HPV16 occurs. PMID:23536685
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Drosophila transcription factor Tramtrack69 binds MEP1 to recruit the chromatin remodeler NuRD.
Reddy, B Ashok; Bajpe, Prashanth Kumar; Bassett, Andrew; Moshkin, Yuri M; Kozhevnikova, Elena; Bezstarosti, Karel; Demmers, Jeroen A A; Travers, Andrew A; Verrijzer, C Peter
2010-11-01
ATP-dependent chromatin-remodeling complexes (remodelers) are essential regulators of chromatin structure and gene transcription. How remodelers can act in a gene-selective manner has remained enigmatic. A yeast two-hybrid screen for proteins binding the Drosophila transcription factor Tramtrack69 (TTK69) identified MEP1. Proteomic characterization revealed that MEP1 is a tightly associated subunit of the NuRD remodeler, harboring the Mi2 enzymatic core ATPase. In addition, we identified the fly homolog of human Deleted in oral cancer 1 (DOC1), also known as CDK2-associated protein 1 (CDK2AP1), as a bona fide NuRD subunit. Biochemical and genetic assays supported the functional association between MEP1, Mi2, and TTK69. Genomewide expression analysis established that TTK69, MEP1, and Mi2 cooperate closely to control transcription. The TTK69 transcriptome profile correlates poorly with remodelers other than NuRD, emphasizing the selectivity of remodeler action. On the genes examined, TTK69 is able to bind chromatin in the absence of NuRD, but targeting of NuRD is dependent on TTK69. Thus, there appears to be a hierarchical relationship in which transcription factor binding precedes remodeler recruitment.
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Fragale, Alessandra; Tartaglia, Marco; Wu, Jie; Gelb, Bruce D
2004-03-01
Noonan syndrome is a developmental disorder with dysmorphic facies, short stature, cardiac defects, and skeletal anomalies, which can be caused by missense PTPN11 mutations. PTPN11 encodes Src homology 2 domain-containing tyrosine phosphatase 2 (SHP2 or SHP-2), a protein tyrosine phosphatase that acts in signal transduction downstream to growth factor, hormone, and cytokine receptors. We compared the functional effects of three Noonan syndrome-causative PTPN11 mutations on SHP2's phosphatase activity, interaction with a binding partner, and signal transduction. All SHP2 mutants had significantly increased basal phosphatase activity compared to wild type, but that activity varied significantly between mutants and was further increased after epidermal growth factor stimulation. Cells expressing SHP2 mutants had prolonged extracellular signal-regulated kinase 2 activation, which was ligand-dependent. Binding of SHP2 mutants to Grb2-associated binder-1 was increased and sustained, and tyrosine phosphorylation of both proteins was prolonged. Coexpression of Grb2-associated binder-1-FF, which lacks SHP2 binding motifs, blocked the epidermal growth factor-mediated increase in SHP2's phosphatase activity and resulted in a dramatic reduction of extracellular signal-regulated kinase 2 activation. Taken together, these results document that Noonan syndrome-associated PTPN11 mutations increase SHP2's basal phosphatase activity, with greater activation when residues directly involved in binding at the interface between the N-terminal Src homology 2 and protein tyrosine phosphatase domains are altered. The SHP2 mutants prolonged signal flux through the RAS/mitogen-activated protein kinase (ERK2/MAPK1) pathway in a ligand-dependent manner that required docking through Grb2-associated binder-1 (GAB1), leading to increased cell proliferation. Copyright 2004 Wiley-Liss, Inc.
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Hale, Sarah J.; Lovell, Simon C.; de Keyzer, Jeanine; Stirling, Colin J.
2010-01-01
Kar2p, an essential Hsp70 chaperone in the endoplasmic reticulum of Saccharomyces cerevisiae, facilitates the transport and folding of nascent polypeptides within the endoplasmic reticulum lumen. The chaperone activity of Kar2p is regulated by its intrinsic ATPase activity that can be stimulated by two different nucleotide exchange factors, namely Sil1p and Lhs1p. Here, we demonstrate that the binding requirements for Lhs1p are complex, requiring both the nucleotide binding domain plus the linker domain of Kar2p. In contrast, the IIB domain of Kar2p is sufficient for binding of Sil1p, and point mutations within IIB specifically blocked Sil1p-dependent activation while remaining competent for activation by Lhs1p. Taken together, these results demonstrate that the interactions between Kar2p and its two nucleotide exchange factors can be functionally resolved and are thus mechanistically distinct. PMID:20430899
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RNA binding specificity of Ebola virus transcription factor VP30.
Schlereth, Julia; Grünweller, Arnold; Biedenkopf, Nadine; Becker, Stephan; Hartmann, Roland K
2016-09-01
The transcription factor VP30 of the non-segmented RNA negative strand Ebola virus balances viral transcription and replication. Here, we comprehensively studied RNA binding by VP30. Using a novel VP30:RNA electrophoretic mobility shift assay, we tested truncated variants of 2 potential natural RNA substrates of VP30 - the genomic Ebola viral 3'-leader region and its complementary antigenomic counterpart (each ∼155 nt in length) - and a series of other non-viral RNAs. Based on oligonucleotide interference, the major VP30 binding region on the genomic 3'-leader substrate was assigned to the internal expanded single-stranded region (∼ nt 125-80). Best binding to VP30 was obtained with ssRNAs of optimally ∼ 40 nt and mixed base composition; underrepresentation of purines or pyrimidines was tolerated, but homopolymeric sequences impaired binding. A stem-loop structure, particularly at the 3'-end or positioned internally, supports stable binding to VP30. In contrast, dsRNA or RNAs exposing large internal loops flanked by entirely helical arms on both sides are not bound. Introduction of a 5´-Cap(0) structure impaired VP30 binding. Also, ssDNAs bind substantially weaker than isosequential ssRNAs and heparin competes with RNA for binding to VP30, indicating that ribose 2'-hydroxyls and electrostatic contacts of the phosphate groups contribute to the formation of VP30:RNA complexes. Our results indicate a rather relaxed RNA binding specificity of filoviral VP30, which largely differs from that of the functionally related transcription factor of the Paramyxoviridae which binds to ssRNAs as short as 13 nt with a preference for oligo(A) sequences.
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Asnani, Mukta; Pestova, Tatyana V.; Hellen, Christopher U.T.
2016-01-01
The cadicivirus IRES diverges structurally from canonical Type 1 IRESs (e.g. poliovirus) but nevertheless also contains an essential GNRA tetraloop in a subdomain (d10c) that is homologous to poliovirus dIVc. In addition to canonical initiation factors, the canonical Type 1 and divergent cadicivirus IRESs require the same IRES trans-acting factor, poly(C)-binding protein 2 (PCBP2). PCBP2 has three KH domains and binds poliovirus IRES domain dIV in the vicinity of the tetraloop. How PCBP2 binds the cadicivirus IRES, and the roles of PCBP2 and the tetraloop in Type 1 IRES function are unknown. Here, directed hydroxyl radical probing showed that KH1 also binds near the cadicivirus tetraloop. KH2 and KH3 bind adjacently to an IRES subdomain (d10b) that is unrelated to dIV, with KH3 in an inverted orientation. KH3 is critical for PCBP2's binding to this IRES whereas KH1 is essential for PCBP2's function in promoting initiation. PCBP2 enforced the wild-type structure of d10c when it contained minor destabilizing substitutions, exposing the tetraloop. Strikingly, PCBP2 enhanced initiation on mutant IRESs that retained consensus GNRA tetraloops, whereas mutants with divergent sequences did not respond to PCBP2. These studies show that PCBP2 enables the IRES to exploit the GNRA tetraloop to enhance initiation. PMID:27387282
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Agarkar, Vinod B.; Babayeva, Nigar D.; Rizzino, Angie
2010-10-08
Ets proteins are transcription factors that activate or repress the expression of genes that are involved in various biological processes, including cellular proliferation, differentiation, development, transformation and apoptosis. Like other Ets-family members, Elf3 functions as a sequence-specific DNA-binding transcriptional factor. A mouse Elf3 C-terminal fragment (amino-acid residues 269-371) containing the DNA-binding domain has been crystallized in complex with mouse type II TGF-{beta} receptor promoter (TR-II) DNA. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 42.66, b = 52, c = 99.78 {angstrom}, and diffracted to a resolution of 2.2 {angstrom}.
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Probst, Olivia C.; Karayel, Evren; Schida, Nicole; Nimmerfall, Elisabeth; Hehenberger, Elisabeth; Puxbaum, Verena; Mach, Lukas
2013-01-01
The M6P (mannose 6-phosphate)/IGF2R (insulin-like growth factor II receptor) interacts with a variety of factors that impinge on tumour invasion and metastasis. It has been shown that expression of wild-type M6P/IGF2R reduces the tumorigenic and invasive properties of receptor-deficient SCC-VII squamous cell carcinoma cells. We have now used mutant forms of M6P/IGF2R to assess the relevance of the different ligand-binding sites of the receptor for its biological activities in this cellular system. The results of the present study demonstrate that M6P/IGF2R does not require a functional binding site for insulin-like growth factor II for inhibition of anchorage-independent growth and matrix invasion by SCC-VII cells. In contrast, the simultaneous mutation of both M6P-binding sites is sufficient to impair all cellular functions of the receptor tested. These findings highlight that the interaction between M6P/IGF2R and M6P-modified ligands is not only important for intracellular accumulation of lysosomal enzymes and formation of dense lysosomes, but is also crucial for the ability of the receptor to suppress SCC-VII growth and invasion. The present study also shows that some of the biological activities of M6P/IGF2R in SCC-VII cells strongly depend on a functional M6P-binding site within domain 3, thus providing further evidence for the non-redundant cellular functions of the individual carbohydrate-binding domains of the receptor. PMID:23347038
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McDonald, Caleb B.; Seldeen, Kenneth L.; Deegan, Brian J.; Bhat, Vikas; Farooq, Amjad
2010-01-01
A ubiquitous component of cellular signaling machinery, Gab1 docker plays a pivotal role in routing extracellular information in the form of growth factors and cytokines to downstream targets such as transcription factors within the nucleus. Here, using isothermal titration calorimetry (ITC) in combination with macromolecular modeling (MM), we show that although Gab1 contains four distinct RXXK motifs, designated G1, G2, G3 and G4, only G1 and G2 motifs bind to the cSH3 domain of Grb2 adaptor and do so with distinct mechanisms. Thus, while the G1 motif strictly requires the PPRPPKP consensus sequence for high-affinity binding to the cSH3 domain, the G2 motif displays preference for the PXVXRXLKPXR consensus. Such sequential differences in the binding of G1 and G2 motifs arise from their ability to adopt distinct polyproline type II (PPII)- and 310-helical conformations upon binding to the cSH3 domain, respectively. Collectively, our study provides detailed biophysical insights into a key protein-protein interaction involved in a diverse array of signaling cascades central to health and disease. PMID:21472810
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Xu, Shuping; Hori, Roderick T
2004-09-01
RNA polymerase I transcription in human cells requires Selectivity Factor 1, a multisubunit complex composed of the TATA-box-binding protein (TBP) and three TBP-associated factors (TAFs) called TAF(I)48, TAF(I)63 and TAF(I)110. Each of the Selectivity Factor 1 subunits binds directly to the other three components, but these interactions have not been characterized. This study is the initial identification and analysis of a TBP-binding domain within a Selectivity Factor 1 TAF. The interaction between human TBP and human TAF(I)48 was initially examined using the yeast two-hybrid assay, and a TBP-binding domain was identified in the carboxyl-terminus of human (h)TAF(I)48. Consistent with this result, the hTAF(I)48 carboxyl-terminus was able to bind directly to TBP in protein-protein interaction assays. When mutations were introduced into the hTAF(I)48 carboxyl-terminus, we identified changes in uncharged and positive residues that affect its interaction with TBP. By examining TBP mutants, residues within and adjacent to helix 2 of TBP, previously demonstrated to interact with subunits of other TBP-containing complexes [Transcription Factor IID (TFIID) and TFIIIB] were also found to diminish its affinity for the carboxyl-terminus of hTAF(I)48. The regions of hTAF(I)48 and TBP that interact are compared to those identified within other complexes containing TBP.
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Szláma, György; Trexler, Mária; Patthy, László
2013-01-01
Myostatin, a negative regulator of skeletal muscle growth, is produced from myostatin precursor by multiple steps of proteolytic processing. After cleavage by a furin-type protease, the propeptide and growth factor domains remain associated, forming a noncovalent complex, the latent myostatin complex. Mature myostatin is liberated from latent myostatin by bone morphogenetic protein 1/tolloid proteases. Here, we show that, in reporter assays, latent myostatin preparations have significant myostatin activity, as the noncovalent complex dissociates at an appreciable rate, and both mature and semilatent myostatin (a complex in which the dimeric growth factor domain interacts with only one molecule of myostatin propeptide) bind to myostatin receptor. The interaction of myostatin receptor with semilatent myostatin is efficiently blocked by WAP, Kazal, immunoglobulin, Kunitz and NTR domain-containing protein 1 or growth and differentiation factor-associated serum protein 2 (WFIKKN1), a large extracellular multidomain protein that binds both mature myostatin and myostatin propeptide [Kondás et al. (2008) J Biol Chem 283, 23677–23684]. Interestingly, the paralogous protein WAP, Kazal, immunoglobulin, Kunitz and NTR domain-containing protein 2 or growth and differentiation factor-associated serum protein 1 (WFIKKN2) was less efficient than WFIKKN1 as an antagonist of the interactions of myostatin receptor with semilatent myostatin. Our studies have shown that this difference is attributable to the fact that only WFIKKN1 has affinity for the propeptide domain, and this interaction increases its potency in suppressing the receptor-binding activity of semilatent myostatin. As the interaction of WFIKKN1 with various forms of myostatin permits tighter control of myostatin activity until myostatin is liberated from latent myostatin by bone morphogenetic protein 1/tolloid proteases, WFIKKN1 may have greater potential as an antimyostatic agent than WFIKKN2. Structured digital abstract Furin cleaves Promyostatin by protease assay (View interaction) myostatin binds to PRO by surface plasmon resonance (View interaction) BMP-1 cleaves Promyostatin by protease assay (View interaction) ACR IIB physically interacts with Latent Myostatin by surface plasmon resonance (View interaction) Promyostatin and Promyostatin bind by comigration in gel electrophoresis (View interaction) WFIKKN1 binds to Latent Myostatin by pull down (View interaction) ACR IIB binds to Mature Myostatin by surface plasmon resonance (View Interaction: 1, 2, 3) WFIKKN1 binds to Myostatin Prodomain by surface plasmon resonance (View Interaction: 1, 2, 3) PMID:23829672
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NASA Astrophysics Data System (ADS)
Ogura, Kenji; Okamura, Hideyasu
2013-10-01
Growth factor receptor-bound protein 2 (Grb2) is a small adapter protein composed of a single SH2 domain flanked by two SH3 domains. The N-terminal SH3 (nSH3) domain of Grb2 binds a proline-rich region present in the guanine nucleotide releasing factor, son of sevenless (Sos). Using NMR relaxation dispersion and chemical shift analysis methods, we investigated the conformational change of the Sos-derived proline-rich peptide during the transition between the free and Grb2 nSH3-bound states. The chemical shift analysis revealed that the peptide does not present a fully random conformation but has a relatively rigid structure. The relaxation dispersion analysis detected conformational exchange of several residues of the peptide upon binding to Grb2 nSH3.
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Stalder, Danièle; Novick, Peter J.
2016-01-01
Sec2p is a guanine nucleotide exchange factor that activates Sec4p, the final Rab GTPase of the yeast secretory pathway. Sec2p is recruited to secretory vesicles by the upstream Rab Ypt32p acting in concert with phosphatidylinositol-4-phosphate (PI(4)P). Sec2p also binds to the Sec4p effector Sec15p, yet Ypt32p and Sec15p compete against each other for binding to Sec2p. We report here that the redundant casein kinases Yck1p and Yck2p phosphorylate sites within the Ypt32p/Sec15p binding region and in doing so promote binding to Sec15p and inhibit binding to Ypt32p. We show that Yck2p binds to the autoinhibitory domain of Sec2p, adjacent to the PI(4)P binding site, and that addition of PI(4)P inhibits Sec2p phosphorylation by Yck2p. Loss of Yck1p and Yck2p function leads to accumulation of an intracellular pool of the secreted glucanase Bgl2p, as well as to accumulation of Golgi-related structures in the cytoplasm. We propose that Sec2p is phosphorylated after it has been recruited to secretory vesicles and the level of PI(4)P has been reduced. This promotes Sec2p function by stimulating its interaction with Sec15p. Finally, Sec2p is dephosphorylated very late in the exocytic reaction to facilitate recycling. PMID:26700316
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Li, Xiaodong; Hoeppner, Luke H.; Jensen, Eric D.; Gopalakrishnan, Rajaram; Westendorf, Jennifer J.
2013-01-01
Runx proteins are essential for a number of developmental processes and are aberrantly expressed in many human cancers. Runx factors bind DNA and co-factors to activate or repress genes crucial for bone formation, hematopoiesis, and neuronal development. Co-activator activator (CoAA) is a nuclear protein that regulates gene expression, RNA splicing and is overexpressed in many human tumors. In this study, we identified CoAA as a Runx2 binding protein. CoAA repressed Runx factor-dependent activation of reporter genes in a histone deacetylase-independent manner. CoAA also blocked Runx2-mediated repression of the Axin2 promoter, a novel Runx target gene. The carboxy-terminus of CoAA is essential for binding the Runt domains of Runx1 and Runx2. In electophoretic mobility shift assays, CoAA inhibited Runx2 interactions with DNA. These data indicate that CoAA is an inhibitor of Runx factors and can negate Runx factor regulation of gene expression. CoAA is expressed at high levels in human fetal osteoblasts and osteosarcoma cell lines. Suppression of CoAA expression by RNA interference reduced osteosarcoma cell viability in vitro, suggesting that it contributes to the proliferation and/or survival of osteoblast lineage cells. PMID:19585539
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van der Does, H. Charlotte; Schmidt, Sarah M.; Langereis, Léon; Hughes, Timothy R.
2016-01-01
Proteins secreted by pathogens during host colonization largely determine the outcome of pathogen-host interactions and are commonly called ‘effectors’. In fungal plant pathogens, coordinated transcriptional up-regulation of effector genes is a key feature of pathogenesis and effectors are often encoded in genomic regions with distinct repeat content, histone code and rate of evolution. In the tomato pathogen Fusarium oxysporum f. sp. lycopersici (Fol), effector genes reside on one of four accessory chromosomes, known as the ‘pathogenicity’ chromosome, which can be exchanged between strains through horizontal transfer. The three other accessory chromosomes in the Fol reference strain may also be important for virulence towards tomato. Expression of effector genes in Fol is highly up-regulated upon infection and requires Sge1, a transcription factor encoded on the core genome. Interestingly, the pathogenicity chromosome itself contains 13 predicted transcription factor genes and for all except one, there is a homolog on the core genome. We determined DNA binding specificity for nine transcription factors using oligonucleotide arrays. The binding sites for homologous transcription factors were highly similar, suggesting that extensive neofunctionalization of DNA binding specificity has not occurred. Several DNA binding sites are enriched on accessory chromosomes, and expression of FTF1, its core homolog FTF2 and SGE1 from a constitutive promoter can induce expression of effector genes. The DNA binding sites of only these three transcription factors are enriched among genes up-regulated during infection. We further show that Ftf1, Ftf2 and Sge1 can activate transcription from their binding sites in yeast. RNAseq analysis revealed that in strains with constitutive expression of FTF1, FTF2 or SGE1, expression of a similar set of plant-responsive genes on the pathogenicity chromosome is induced, including most effector genes. We conclude that the Fol pathogenicity chromosome may be partially transcriptionally autonomous, but there are also extensive transcriptional connections between core and accessory chromosomes. PMID:27855160
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Cho, Ok Hyun; Mallappa, Chandrashekara; Hernández-Hernández, J Manuel; Rivera-Pérez, Jaime A; Imbalzano, Anthony N
2015-01-01
Among the complexities of skeletal muscle differentiation is a temporal distinction in the onset of expression of different lineage-specific genes. The lineage-determining factor MyoD is bound to myogenic genes at the onset of differentiation whether gene activation is immediate or delayed. How temporal regulation of differentiation-specific genes is established remains unclear. Using embryonic tissue, we addressed the molecular differences in the organization of the myogenin and muscle creatine kinase (MCK) gene promoters by examining regulatory factor binding as a function of both time and spatial organization during somitogenesis. At the myogenin promoter, binding of the homeodomain factor Pbx1 coincided with H3 hyperacetylation and was followed by binding of co-activators that modulate chromatin structure. MyoD and myogenin binding occurred subsequently, demonstrating that Pbx1 facilitates chromatin remodeling and modification before myogenic regulatory factor binding. At the same time, the MCK promoter was bound by HDAC2 and MyoD, and activating histone marks were largely absent. The association of HDAC2 and MyoD was confirmed by co-immunoprecipitation, proximity ligation assay (PLA), and sequential ChIP. MyoD differentially promotes activated and repressed chromatin structures at myogenic genes early after the onset of skeletal muscle differentiation in the developing mouse embryo. © 2014 Wiley Periodicals, Inc.
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Sybill Amelon; Robert T. Brooks; Jessie Glaeser; Megan Friggens; Daniel Lindner; Susan C. Loeb; Ann Lynch; Drew Minnis; Roger Perry; Mary M. Rowland; Monica Tomosy; Ted Weller
2012-01-01
The National Plan for Assisting States, Federal Agencies, and Tribes in Managing White-Nose Syndrome in Bats (National WNS Plan), is a document prepared jointly by the U.S. Departments of the Interior, Agriculture, and Defense, along with the Association of Fish and Wildlife Agencies. This document provides a strategic framework for the investigation and management of...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-09-17
... issue a special use permit for the proposal. RUS is the lead agency conducting the EIS, and the USFS... refined as part of the EIS scoping process and will be addressed in the EIS. Public health and safety, environmental impacts, and engineering aspects of the proposal will be considered in the EIS. RUS is the lead...
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Assessment of soil disturbance in forests of the interior Columbia River basin: a critique
Richard E. Miller; James D. McIver; Steven W. Howes; William B. Gaeuman
2010-01-01
We present results and inferences from 15 soil-monitoring projects by the USDA Forest Service (USFS) after logging in the interior Columbia River basin. Details and comments about each project are provided in separate appendixes. In general, application of past protocols overestimated the percentage of âdetrimentallyâ disturbed soil in harvested units. Based on this...
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Universal Service Fund: Background and Options for Reform
2010-03-11
pursues violators and initiates enforcement actions including notices of liability, suspensions , consent decrees, and debarments .46 The Department...series of measures to safeguard the USF to deter fraud, waste, and abuse. Included in the measures taken are those that extend the debarment rules...three years) and sanctions for criminal and civil violations beyond the Schools and Libraries Program to cover all four programs; tighten rules
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Effects of Climate Change on Cultural Resources in the Northern Rockies Region [Chapter 12
Carl M. Davis
2018-01-01
People have inhabited the Northern Rocky Mountains of the United States since the close of the last Pleistocene glacial period, some 14,000 years B.P. (Fagan 1990; Meltzer 2009). Evidence of this ancient and more recent human occupation is found throughout the Forest Service, U.S. Department of Agriculture (USFS) Northern Region and the Greater Yellowstone Area,...
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Integrating ecosystem services into national Forest Service policy and operations
Robert Deal; Lisa Fong; Erin Phelps; Emily Weidner; Jonas Epstein; Tommie Herbert; Mary Snieckus; Nikola Smith; Tania Ellersick; Greg Arthaud
2017-01-01
The ecosystem services concept describes the many benefits people receive from nature. It highlights the importance of managing public and private lands sustainably to ensure these benefits continue into the future, and it closely aligns with the U.S. Forest Service (USFS) mission to âsustain the health, diversity, and productivity of the Nationâs forests and...
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Welcome - TampaBay.WaterAtlas.org
An edition of: WaterAtlas.orgPresented By: USF Water Institute Choose a Water Atlas Charlotte Harbor NEP Water Atlas Hillsborough County Water Atlas Lake County Water Atlas Manatee County Water Atlas Orange County Water Atlas Pinellas County Water Atlas Polk County Water Atlas Sarasota County Water Atlas
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Larry E. Laing; David Gori; James T. Jones
2005-01-01
The multi-partner Greater Huachuca Mountains fire planning effort involves over 500,000 acres of public and private lands. This large area supports distinct landscapes that have evolved with fire. Utilizing GIS as a tool, the United States Forest Service (USFS), General Ecosystem Survey (GES), and Natural Resources Conservation Service (NRCS) State Soil Geographic...
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Carbon stocks on forestland of the United States, with emphasis on USDA Forest Service ownership
Linda S. Heath; James E. Smith; Christopher W. Woodall; David L. Azuma; Karen L. Waddell
2011-01-01
The U.S. Department of Agriculture Forest Service (USFS) manages one-fifth of the area of forestland in the United States. The Forest Service Roadmap for responding to climate change identified assessing and managing carbon stocks and change as a major element of its plan. This study presents methods and results of estimating current forest carbon stocks and change in...
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Brian P. Oswald; Ike McWhorter; Penny. Whisenant
2011-01-01
The 13,250-acre Upland Island Wilderness (UIW) in Texas was established in 1984 and is managed by the United States Forest Service (USFS). Historically, portions of it consisted of open and diverse longleaf pine (Pinus palustris) ecosystems which depend on frequent, low-intensity surface fires. As in many other relatively small wilderness areas, the...
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Carbon stocks on forestland of the United States, with emphaisis on USDA Forest Service ownership
Linda S. Heath; James E. Smith; Christopher W. Woodall; Dave Azuma; Karen L. Waddell
2011-01-01
The U.S. Department of Agriculture Forest Service (USFS) manages one-fifth of the area of forestland in the United States. The Forest Service Roadmap for responding to climate change identified assessing and managing carbon stocks and change as a major element of its plan. This study presents methods and results of estimating current forest carbon stocks and change in...
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The Fundamental Chemistry and Physics of Munitions under Extreme Conditions
2011-02-01
I. I. Oleynik, S. V. Zybin, and C. T. White, “Density Functional Theory Calculations of Solid Nitromethane under Hydrostatic and Uniaxial...White (NRL), Ivan Oleynik (USF): anisotropic nonlinear elasticity and equations of states of crystalline EM (PETN, RDX, HMX, TATB, nitromethane ...and nitromethane ; Aidan Thompson (SNL): shock-to-detonation transition in PETN and CL-20; Ronnie Kosloff (Hebrew University of Jerusalem
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Karen Schleeweis; Chengquan Huang; Khaldoun Rishmawi; Feng Aron Zhao; Jeffery G. Masek; Richard K. Houghton; Samuel N. Goward
2015-01-01
For nearly a decade, the USFS FIA, NASA, and the University of Maryland have collaborated on the NASA/NACP funded North American Forest Dynamics (NAFD) project, and developed new approaches for annual mapping of CONUS forest dynamics (1984-2011). Building on this foundation of empirical research and results, the collaboration will continue with a new Carbon Cycle...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-09
... affects 59.78 acres of land withdrawn for the Panelli Seed Orchard. The USFS has determined the remaining... Panelli Seed Orchard (56 FR 11940 (1991)), for an additional 20-year term. PLO No. 6874 withdrew certain... Meridian Fremont National Forest Panelli Seed Orchard T. 37 S., R. 15 E., sec. 24, NE\\1/4\\SE\\1/4\\. [[Page...
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Mission Command: Retooling the Leadership Paradigm for Homeland Security Crisis Response?
2015-03-01
security crisis management. While, there is some literature regarding the efforts by the USFS and the federal wildland firefighting community towards...psychologically stressful and potentially overwhelming. The need to communicate and cooperate with other agencies in a crisis places added...environment where formal hierarchies and tightly controlled plans may become irrelevant in a crisis . Grant and De Waard examined the communication
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Alexander Hernandez; Sean P. Healey; Chenquan Huang; R. Douglas Ramsey
2015-01-01
As part of the U.S. Forest Service (USFS), National Forest System (NFS) comprehensive plan for carbon monitoring, a detailed temporal mapping of forest disturbances across all National Forests in the United States has been conducted. A long-term annual time series of data layers that show the timing, extent, type, and magnitude of disturbance beginning in 1990 and...
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2000-08-01
management for NPS. The State nonpoint Source Task Force coordinates joint watershed management efforts with SCS, USFS, BLM. Intense grazing and...nonpoint source water pollution discharges from unimproved lands, particularly military lands. Increasing emphasis at national and state levels on...lands, particularly military lands. Increasing emphasis at national and state levels on controlling pollutant discharges from nonpoint sources and
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ERIC Educational Resources Information Center
Micceri, Theodore
2005-01-01
This study sought to determine whether consistent differences in enrollment and graduation among different racial/ethnic and sex groups occur at different colleges of Engineering in the Florida State University System (SUS). Analyses were limited to the major institutions (UF, FSU, USF, UCF, FAU, FIU) with the addition of FAMU due to a high…
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How useful is LiDAR in establishing a stream gauging network in a tropical experimental forest
Boris Poff; Daniel G. Neary; Gregory P. Asner
2008-01-01
In the late summer of 2007 the Institute for Pacific Islands Forestry (IPIF), which is part of the US Forest Service Pacific Southwestern Research Station, asked the USFS Rocky Mountain Research Station's (RMRS) Air, Water and Aquatic Program's (AWA) Southwest Watershed Science Team for assistance in the establishing baseline data in the initial phase of a...
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ERIC Educational Resources Information Center
Twitchell, Anne; Sprehn, Mary
An evaluation of the Ohio College Library Center's (OCLC) proposed Serials Control Subsystem was undertaken to determine what effect the system would have on the operation of the Serials Department at the University of South Florida (USF) Library. The system would consist of three components: 1) claiming--identifying missing issues and generating…
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DOT National Transportation Integrated Search
2007-07-24
A review of the transportation in the Front Range region of Colorado by the inter-agency : Transportation Assistance Group (TAG) was conducted July 24-26, 2007, on behalf of : the U.S. Department of Agriculture Forest Service (USFS) in cooperation wi...
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Clarifying details on a 1930s-era pine-hardwood stand in Arkansas
Don C. Bragg
2015-01-01
Data from recently discovered daily-work logs of US Forest Service (USFS) researcher Russell R. Reynolds enabled me to clarify a study I published a decade ago on a 1930s-vintage unmanaged, second-growth Pinus (pine)âhardwood stand in southeastern Arkansas. Though still too vague to reveal every detail, Reynoldsâ work logs confirmed a number of...
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Keith Stockmann; Nathaniel Anderson; Jesse Young; Ken Skog; Sean Healey; Dan Loeffler; Edward Butler; J. Greg Jones; James Morrison
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Edward Butler; Keith Stockmann; Nathaniel Anderson; Ken Skog; Sean Healey; Dan Loeffler; J. Greg Jones; James Morrison; Jesse Young
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Keith Stockmann; Nathaniel Anderson; Jesse Young; Ken Skog; Sean Healey; Dan Loeffler; Edward Butler; J. Greg Jones; James Morrison
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Dan Loeffler; Nathaniel Anderson; Keith Stockmann; Ken Skog; Sean Healey; J. Greg Jones; James Morrison; Jesse Young
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Keith Stockmann; Nathaniel Anderson; Jesse Young; Ken Skog; Sean Healey; Dan Loeffler; Edward Butler; J. Greg Jones; James Morrison
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Environmental accounting of natural capital and ecosystem services for the US National Forest System
Elliot T. Campbell; Mark T. Brown; NO-VALUE
2012-01-01
The National Forests of the United States encompass 192.7 million acres (78 million hectares) of land, which is nearly five percent of the total land area of the nation. These lands are managed by the US Forest Service (USFS) for multiple uses, including extraction of timber, production of fossil fuels and minerals, public recreation, and the preservation of...
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Dan Loeffler; Nathaniel Anderson; Keith Stockmann; Ken Skog; Sean Healey; J. Greg Jones; James Morrison; Jesse Young
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Keith Stockmann; Nathaniel Anderson; Jesse Young; Ken Skog; Sean Healey; Dan Loeffler; Edward Butler; J. Greg Jones; James Morrison
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Edward Butler; Keith Stockmann; Nathaniel Anderson; Jesse Young; Ken Skog; Sean Healey; Dan Loeffler; J. Greg Jones; James Morrison
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Dan Loeffler; Nathaniel Anderson; Keith Stockmann; Ken Skog; Sean Healey; J. Greg Jones; James Morrison; Jesse Young
2014-01-01
Global forests capture and store significant amounts of carbon through photosynthesis. When carbon is removed from forests through harvest, a portion of the harvested carbon is stored in wood products, often for many decades. The United States Forest Service (USFS) and other agencies are interested in accurately accounting for carbon flux associated with harvested wood...
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Jill Grenon; Terry Svalberg; Ted Porwoll; Mark Story
2010-01-01
Air quality monitoring data from several programs in and around the Bridger-Teton (B-T) National Forest - National Atmospheric Deposition Program (NADP), longterm lake monitoring, long-term bulk precipitation monitoring (both snow and rain), and Interagency Monitoring of Protected Visual Environments (IMPROVE) - were analyzed in this report. Trends were analyzed using...
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Radiotelemetry Tracking at Lake Conway, Florida.
1982-08-01
USF) initiated a study to evaluate the impact of the introduced white amur on the resident herpetofaunal community . A major part of this effort was...initiated a study to determine the impact of the introduced white amur on the resident herpetofaunal community (Godley, in preparation). A major part of...42. Analysis of the project funding schedule indicated that herpetofaunal radiotagging would be economically feasible only if the existing mobile
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Matthew Wibbenmeyer; Michael Hand; David Calkin
2012-01-01
The U.S. Department of Agriculture, Forest Service (USFS) has, in recent years, increasingly emphasized the importance of safety to its employees, but wildfire management remains a risky endeavor. While wildfire management decisions affecting safety and exposure of firefighters to the wildland fire environment may be aided by decision support tools such the Wildfire...
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Huebert, Dana J.; Kuan, Pei-Fen; Keleş, Sündüz
2012-01-01
The response to stressful stimuli requires rapid, precise, and dynamic gene expression changes that must be coordinated across the genome. To gain insight into the temporal ordering of genome reorganization, we investigated dynamic relationships between changing nucleosome occupancy, transcription factor binding, and gene expression in Saccharomyces cerevisiae yeast responding to oxidative stress. We applied deep sequencing to nucleosomal DNA at six time points before and after hydrogen peroxide treatment and revealed many distinct dynamic patterns of nucleosome gain and loss. The timing of nucleosome repositioning was not predictive of the dynamics of downstream gene expression change but instead was linked to nucleosome position relative to transcription start sites and specific cis-regulatory elements. We measured genome-wide binding of the stress-activated transcription factor Msn2p over time and found that Msn2p binds different loci with different dynamics. Nucleosome eviction from Msn2p binding sites was common across the genome; however, we show that, contrary to expectation, nucleosome loss occurred after Msn2p binding and in fact required Msn2p. This negates the prevailing model that nucleosomes obscuring Msn2p sites regulate DNA access and must be lost before Msn2p can bind DNA. Together, these results highlight the complexities of stress-dependent chromatin changes and their effects on gene expression. PMID:22354995
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NASA Astrophysics Data System (ADS)
Hernandez, A. J.
2015-12-01
The Landsat archive is increasingly being used to detect trends in the occurrence of forest disturbance. Beyond information about the amount of area affected, forest managers need to know if and how disturbance regimes change. The National Forest System (NFS) has developed a comprehensive plan for carbon monitoring that requires a detailed temporal mapping of forest disturbances across 75 million hectares. A long-term annual time series that shows the timing, extent, and type of disturbance beginning in 1990 and ending in 2011 has been prepared for several USFS Regions, including the Northern Region. Our mapping starts with an automated detection of annual disturbances using a time series of historical Landsat imagery. Automated detections are meticulously inspected, corrected and labeled using various USFS ancillary datasets. The resulting maps of verified disturbance show the timing and types are fires, harvests, insect activity, disease, and abiotic (wind, drought, avalanche) damage. Also, the magnitude of each change event is modeled in terms of the proportion of canopy cover lost. The sequence of disturbances for every pixel since 1990 has been consistently mapped and is available across the entirety of NFS. Our datasets contain sufficient information to describe the frequency of stand replacement, as well as how often disturbance results in only a partial loss of canopy. This information provides empirical insight into how an initial disturbance may predispose a stand to further disturbance, and it also show a climatic signal in the occurrence of processes such as fire and insect epidemics. Thus, we have the information to model the likelihood of occurrence of certain disturbances after a given event (i.e. if we have a fire in the past what does that do to the likelihood of occurrence of insects in the future). Here, we explore if previous disturbance history is a reliable predictor of additional disturbance in the future and we present results of applying logistic regression to obtain predicted probabilities of occurrence of additional disturbance types. We describe responses in additional disturbance and prominent trends for each major forest type.
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Patikoglou, Georgia A; Westblade, Lars F; Campbell, Elizabeth A; Lamour, Valérie; Lane, William J; Darst, Seth A
2007-09-21
The Escherichia coli Rsd protein binds tightly and specifically to the RNA polymerase (RNAP) sigma(70) factor. Rsd plays a role in alternative sigma factor-dependent transcription by biasing the competition between sigma(70) and alternative sigma factors for the available core RNAP. Here, we determined the 2.6 A-resolution X-ray crystal structure of Rsd bound to sigma(70) domain 4 (sigma(70)(4)), the primary determinant for Rsd binding within sigma(70). The structure reveals that Rsd binding interferes with the two primary functions of sigma(70)(4), core RNAP binding and promoter -35 element binding. Interestingly, the most highly conserved Rsd residues form a network of interactions through the middle of the Rsd structure that connect the sigma(70)(4)-binding surface with three cavities exposed on distant surfaces of Rsd, suggesting functional coupling between sigma(70)(4) binding and other binding surfaces of Rsd, either for other proteins or for as yet unknown small molecule effectors. These results provide a structural basis for understanding the role of Rsd, as well as its ortholog, AlgQ, a positive regulator of Pseudomonas aeruginosa virulence, in transcription regulation.
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Crystal structure of the Escherichia coli regulator of σ70, Rsd, in complex with σ70 domain 4
Patikoglou, Georgia A.; Westblade, Lars F.; Campbell, Elizabeth A.; Lamour, Valérie; Lane, William J.; Darst, Seth A.
2007-01-01
Summary The Escherichia coli Rsd protein binds tightly and specifically to the RNA polymerase (RNAP) σ70 factor. Rsd plays a role in alternative σ factor-dependent transcription by biasing the competition between σ70 and alternative σ factors for the available core RNAP. Here, we determined the 2.6 Å-resolution X-ray crystal structure of Rsd bound to σ70 domain 4 (σ704), the primary determinant for Rsd binding within σ70. The structure reveals that Rsd binding interferes with the two primary functions of σ704, core RNAP binding and promoter –35 element binding. Interestingly, the most highly conserved Rsd residues form a network of interactions through the middle of the Rsd structure that connect the σ704-binding surface with three cavities exposed on distant surfaces of Rsd, suggesting functional coupling between σ704 binding and other binding surfaces of Rsd, either for other proteins or for as yet unknown small molecule effectors. These results provide a structural basis for understanding the role of Rsd, as well as its ortholog, AlgQ, a positive regulator of Pseudomonas aeruginosa virulence, in transcription regulation. PMID:17681541
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Patikoglou,G.; Westblade, L.; Campbell, E.
The Escherichia coli Rsd protein binds tightly and specifically to the RNA polymerase (RNAP) {sigma}{sup 70} factor. Rsd plays a role in alternative {sigma} factor-dependent transcription by biasing the competition between {sigma}{sup 70} and alternative {sigma} factors for the available core RNAP. Here, we determined the 2.6 {angstrom}-resolution X-ray crystal structure of Rsd bound to {sigma}{sup 70} domain 4 ({sigma}{sup 70}{sub 4}), the primary determinant for Rsd binding within {sigma}{sup 70}. The structure reveals that Rsd binding interferes with the two primary functions of {sigma}{sup 70}{sub 4}, core RNAP binding and promoter -35 element binding. Interestingly, the most highly conservedmore » Rsd residues form a network of interactions through the middle of the Rsd structure that connect the {sigma}{sup 70}{sub 4}-binding surface with three cavities exposed on distant surfaces of Rsd, suggesting functional coupling between {sigma}{sup 70}{sub 4} binding and other binding surfaces of Rsd, either for other proteins or for as yet unknown small molecule effectors. These results provide a structural basis for understanding the role of Rsd, as well as its ortholog, AlgQ, a positive regulator of Pseudomonas aeruginosa virulence, in transcription regulation.« less
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Crystallographic analysis of CD40 recognition and signaling by human TRAF2
McWhirter, Sarah M.; Pullen, Steven S.; Holton, James M.; Crute, James J.; Kehry, Marilyn R.; Alber, Tom
1999-01-01
Tumor necrosis factor receptor superfamily members convey signals that promote diverse cellular responses. Receptor trimerization by extracellular ligands initiates signaling by recruiting members of the tumor necrosis factor receptor-associated factor (TRAF) family of adapter proteins to the receptor cytoplasmic domains. We report the 2.4-Å crystal structure of a 22-kDa, receptor-binding fragment of TRAF2 complexed with a functionally defined peptide from the cytoplasmic domain of the CD40 receptor. TRAF2 forms a mushroom-shaped trimer consisting of a coiled coil and a unique β-sandwich domain. Both domains mediate trimerization. The CD40 peptide binds in an extended conformation with every side chain in contact with a complementary groove on the rim of each TRAF monomer. The spacing between the CD40 binding sites on TRAF2 supports an elegant signaling mechanism in which trimeric, extracellular ligands preorganize the receptors to simultaneously recognize three sites on the TRAF trimer. PMID:10411888
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In Vivo Chromatin Targets of the Transcription Factor Yin Yang 2 in Trophoblast Stem Cells
Pérez-Palacios, Raquel; Macías-Redondo, Sofía; Climent, María; Contreras-Moreira, Bruno; Muniesa, Pedro; Schoorlemmer, Jon
2016-01-01
Background Yin Yang 2 (YY2) is a zinc finger protein closely related to the well-characterized Yin Yang 1 (YY1). YY1 is a DNA-binding transcription factor, with defined functions in multiple developmental processes, such as implantation, cell differentiation, X inactivation, imprinting and organogenesis. Yy2 has been treated as a largely immaterial duplication of Yy1, as they share high homology in the Zinc Finger-region and similar if not identical in vitro binding sites. In contrast to these similarities, gene expression alterations in HeLa cells with attenuated levels of either Yy1 or Yy2 were to some extent gene-specific. Moreover, the chromatin binding sites for YY2, except for its association with transposable retroviral elements (RE) and Endogenous Retroviral Elements (ERVs), remain to be identified. As a first step towards defining potential Yy2 functions matching or complementary to Yy1, we considered in vivo DNA binding sites of YY2 in trophoblast stem (TS) cells. Results We report the presence of YY2 protein in mouse-derived embryonic stem (ES) and TS cell lines. Following up on our previous report on ERV binding by YY2 in TS cells, we investigated the tissue-specificity of REX1 and YY2 binding and confirm binding to RE/ERV targets in both ES cells and TS cells. Because of the higher levels of expression, we chose TS cells to understand the role of Yy2 in gene and chromatin regulation. We used in vivo YY2 association as a measure to identify potential target genes. Sequencing of chromatin obtained in chromatin-immunoprecipitation (ChIP) assays carried out with αYY2 serum allowed us to identify a limited number of chromatin targets for YY2. Some putative binding sites were validated in regular ChIP assays and gene expression of genes nearby was altered in the absence of Yy2. Conclusions YY2 binding to ERVs is not confined to TS cells. In vivo binding sites share the presence of a consensus binding motif. Selected sites were uniquely bound by YY2 as opposed to YY1, suggesting that YY2 exerts unique contributions to gene regulation. YY2 binding was not generally associated with gene promoters. However, several YY2 binding sites are linked to long noncoding RNA (lncRNA) genes and we show that the expression levels of a few of those are Yy2-dependent. PMID:27191592
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Han, S H; Yea, S S; Jeon, Y J; Yang, K H; Kaminski, N E
1998-12-01
Transforming growth factor beta1 (TGF-beta1) has been previously shown to modulate interleukin 2 (IL-2) secretion by activated T-cells. In the present studies, we determined that TGF-beta1 induced IL-2 mRNA expression in the murine T-cell line EL4, in the absence of other stimuli. IL-2 mRNA expression was significantly induced by TGF-beta1 (0.1-1 ng/ml) over a relatively narrow concentration range, which led to the induction of IL-2 secretion. Under identical condition, we examined the effect of TGF-beta1 on the activity of nuclear factor AT (NF-AT), nuclear factor kappaB (NF-kappaB), activator protein-1 (AP-1) and octamer, all of which contribute to the regulation of IL-2 gene expression. Electrophoretic mobility shift assays showed that TGF-beta1 markedly increased NF-AT, NF-kappaB and AP-1 binding to their respective cognate DNA binding sites, whereas octamer binding remained constant, as compared with untreated cells. Employing a reporter gene expression system with p(NF-kappaB)3-CAT, p(NF-AT)3-CAT and p(AP-1)3-CAT, TGF-beta1 treatment of transfected EL4 cells induced a dose-related increase in chloramphenicol acetyltransferase activity that correlated well with the DNA binding profile found in the electrophoretic mobility shift assay studies. These results show that TGF-beta1, in the absence of any additional stimuli, up-regulates the activity of key transcription factors involved in IL-2 gene expression, including NF-AT, NF-kappaB and AP-1, to help promote IL-2 mRNA expression by EL4 cells.
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Li, Cong; Yue, Jing; Wu, Xiaowei; Xu, Cong; Yu, Jingjuan
2014-10-01
The DREB (dehydration-responsive element binding)-type transcription factors regulate the expression of stress-inducible genes by binding the DRE/CRT cis-elements in promoter regions. The upstream transcription factors that regulate the transcription of DREB transcription factors have not been clearly defined, although the function of DREB transcription factors in abiotic stress is known. In this study, an abscisic acid (ABA)-responsive DREB-binding protein gene (SiARDP) was cloned from foxtail millet (Setaria italica). The transcript level of SiARDP increased not only after drought, high salt, and low temperature stresses, but also after an ABA treatment in foxtail millet seedlings. Two ABA-responsive elements (ABRE1: ACGTGTC; ABRE2: ACGTGGC) exist in the promoter of SiARDP. Further analyses showed that two ABA-responsive element binding (AREB)-type transcription factors, SiAREB1 and SiAREB2, could physically bind to the ABRE core element in vitro and in vivo. The constitutive expression of SiARDP in Arabidopsis thaliana enhanced drought and salt tolerance during seed germination and seedling development, and overexpression of SiARDP in foxtail millet improved drought tolerance. The expression levels of target genes of SiARDP were upregulated in transgenic Arabidopsis and foxtail millet. These results reveal that SiARDP, one of the target genes of SiAREB, is involved in ABA-dependent signal pathways and plays a critical role in the abiotic stress response in plants. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.
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Yu, Tao; Yang, Yanyan; Kwak, Yi-Seong; Song, Gwan Gyu; Kim, Mi-Yeon; Rhee, Man Hee; Cho, Jae Youl
2017-04-01
Ginsenoside Rc (G-Rc) is one of the major protopanaxadiol-type saponins isolated from Panax ginseng , a well-known medicinal herb with many beneficial properties including anticancer, anti-inflammatory, antiobesity, and antidiabetic effects. In this study, we investigated the effects of G-Rc on inflammatory responses in vitro and examined the mechanisms of these effects. The in vitro inflammation system used lipopolysaccharide-treated macrophages, tumor necrosis factor-α/interferon-γ-treated synovial cells, and HEK293 cells transfected with various inducers of inflammation. G-Rc significantly inhibited the expression of macrophage-derived cytokines, such as tumor necrosis factor-α and interleukin-1β. G-Rc also markedly suppressed the activation of TANK-binding kinase 1/IκB kinase ε/interferon regulatory factor-3 and p38/ATF-2 signaling in activated RAW264.7 macrophages, human synovial cells, and HEK293 cells. G-Rc exerts its anti-inflammatory actions by suppressing TANK-binding kinase 1/IκB kinase ε/interferon regulatory factor-3 and p38/ATF-2 signaling.
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Granoff, Dan M.; Costa, Isabella; Konar, Monica; Giuntini, Serena; Van Rompay, Koen K. A.; Beernink, Peter T.
2015-01-01
Background. The meningococcal vaccine antigen, factor H (FH)–binding protein (FHbp), binds human complement FH. In human FH transgenic mice, binding decreased protective antibody responses. Methods. To investigate the effect of primate FH binding, we immunized rhesus macaques with a 4-component serogroup B vaccine (4CMenB). Serum FH in 6 animals bound strongly to FHbp (FHbp-FHhigh) and, in 6 animals, bound weakly to FHbp (FHbp-FHlow). Results. There were no significant differences between the respective serum bactericidal responses of the 2 groups against meningococcal strains susceptible to antibody to the NadA or PorA vaccine antigens. In contrast, anti-FHbp bactericidal titers were 2-fold lower in FHbp-FHhigh macaques against a strain with an exact FHbp match to the vaccine (P = .08) and were ≥4-fold lower against 4 mutants with other FHbp sequence variants (P ≤ .005, compared with FHbp-FHlow macaques). Unexpectedly, postimmunization sera from all 12 macaques enhanced FH binding to meningococci. In contrast, serum anti-FHbp antibodies elicited by 4CMenB in mice whose mouse FH did not bind to the vaccine antigen inhibited FH binding. Conclusions. Binding of FH to FHbp decreases protective anti-FHbp antibody responses of macaques to 4CMenB. Even low levels of FH binding skew the antibody repertoire to FHbp epitopes outside of the FH-binding site, which enhance FH binding. PMID:25676468
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Chen, Rui; Xu, Min; Hogg, Richard T.; Li, Jiwen; Little, Bertis; Gerard, Robert D.; Garcia, Joseph A.
2012-01-01
Hypoxia-inducible factors (HIFs) are oxygen-sensitive transcription factors. HIF-1α plays a prominent role in hypoxic gene induction. HIF-2α target genes are more restricted but include erythropoietin (Epo), one of the most highly hypoxia-inducible genes in mammals. We previously reported that HIF-2α is acetylated during hypoxia but is rapidly deacetylated by the stress-responsive deacetylase Sirtuin 1. We now demonstrate that the lysine acetyltransferases cAMP-response element-binding protein-binding protein (CBP) and p300 are required for efficient Epo induction during hypoxia. However, despite close structural similarity, the roles of CBP and p300 differ in HIF signaling. CBP acetylates HIF-2α, is a major coactivator for HIF-2-mediated Epo induction, and is required for Sirt1 augmentation of HIF-2 signaling during hypoxia in Hep3B cells. In comparison, p300 is a major contributor for HIF-1 signaling as indicated by induction of Pgk1. Whereas CBP can bind with HIF-2α independent of the HIF-2α C-terminal activation domain via enzyme/substrate interactions, p300 only complexes with HIF-2α through the C-terminal activation domain. Maximal CBP/HIF-2 signaling requires intact CBP acetyltransferase activity in both Hep3B cells as well as in mice. PMID:22807441
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Lin,L.
2007-01-01
Lactadherin, a glycoprotein secreted by a variety of cell types, contains two EGF domains and two C domains with sequence homology to the C domains of blood coagulation proteins factor V and factor VIII. Like these proteins, lactadherin binds to phosphatidylserine (PS)-containing membranes with high affinity. We determined the crystal structure of the bovine lactadherin C2 domain (residues 1 to 158) at 2.4 {angstrom}. The lactadherin C2 structure is similar to the C2 domains of factors V and VIII (rmsd of C{sub {alpha}} atoms of 0.9 {angstrom} and 1.2 {angstrom}, and sequence identities of 43% and 38%, respectively). The lactadherinmore » C2 domain has a discoidin-like fold containing two {beta}-sheets of five and three antiparallel {beta}-strands packed against one another. The N and C termini are linked by a disulfide bridge between Cys1 and Cys158. One {beta}-turn and two loops containing solvent-exposed hydrophobic residues extend from the C2 domain {beta}-sandwich core. In analogy with the C2 domains of factors V and VIII, some or all of these solvent-exposed hydrophobic residues, Trp26, Leu28, Phe31, and Phe81, likely participate in membrane binding. The C2 domain of lactadherin may serve as a marker of cell surface phosphatidylserine exposure and may have potential as a unique anti-thrombotic agent.« less
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Lin,L.; Huai, Q.; Huang, M.
2007-01-01
Lactadherin, a glycoprotein secreted by a variety of cell types, contains two EGF domains and two C domains with sequence homology to the C domains of blood coagulation proteins factor V and factor VIII. Like these proteins, lactadherin binds to phosphatidylserine (PS)-containing membranes with high affinity. We determined the crystal structure of the bovine lactadherin C2 domain (residues 1 to 158) at 2.4 Angstroms. The lactadherin C2 structure is similar to the C2 domains of factors V and VIII (rmsd of C? atoms of 0.9 Angstroms and 1.2 Angstroms, and sequence identities of 43% and 38%, respectively). The lactadherin C2more » domain has a discoidin-like fold containing two ?-sheets of five and three antiparallel ?-strands packed against one another. The N and C termini are linked by a disulfide bridge between Cys1 and Cys158. One ?-turn and two loops containing solvent-exposed hydrophobic residues extend from the C2 domain ?-sandwich core. In analogy with the C2 domains of factors V and VIII, some or all of these solvent-exposed hydrophobic residues, Trp26, Leu28, Phe31, and Phe81, likely participate in membrane binding. The C2 domain of lactadherin may serve as a marker of cell surface phosphatidylserine exposure and may have potential as a unique anti-thrombotic agent.« less
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Jadwin, Joshua A; Oh, Dongmyung; Curran, Timothy G; Ogiue-Ikeda, Mari; Jia, Lin; White, Forest M; Machida, Kazuya; Yu, Ji; Mayer, Bruce J
2016-04-12
While the affinities and specificities of SH2 domain-phosphotyrosine interactions have been well characterized, spatio-temporal changes in phosphosite availability in response to signals, and their impact on recruitment of SH2-containing proteins in vivo, are not well understood. To address this issue, we used three complementary experimental approaches to monitor phosphorylation and SH2 binding in human A431 cells stimulated with epidermal growth factor (EGF): 1) phospho-specific mass spectrometry; 2) far-Western blotting; and 3) live cell single-molecule imaging of SH2 membrane recruitment. Far-Western and MS analyses identified both well-established and previously undocumented EGF-dependent tyrosine phosphorylation and binding events, as well as dynamic changes in binding patterns over time. In comparing SH2 binding site phosphorylation with SH2 domain membrane recruitment in living cells, we found in vivo binding to be much slower. Delayed SH2 domain recruitment correlated with clustering of SH2 domain binding sites on the membrane, consistent with membrane retention via SH2 rebinding.
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Weissbach, Julia; Schikora, Franziska; Weber, Anja; Kessels, Michael; Posern, Guido
2016-05-15
The myocardin-related transcription factors (MRTFs) are coactivators of serum response factor (SRF)-mediated gene expression. Activation of MRTF-A occurs in response to alterations in actin dynamics and critically requires the dissociation of repressive G-actin-MRTF-A complexes. However, the mechanism leading to the release of MRTF-A remains unclear. Here we show that WH2 domains compete directly with MRTF-A for actin binding. Actin nucleation-promoting factors, such as N-WASP and WAVE2, as well as isolated WH2 domains, including those of Spire2 and Cobl, activate MRTF-A independently of changes in actin dynamics. Simultaneous inhibition of Arp2-Arp3 or mutation of the CA region only partially reduces MRTF-A activation by N-WASP and WAVE2. Recombinant WH2 domains and the RPEL domain of MRTF-A bind mutually exclusively to cellular and purified G-actin in vitro The competition by different WH2 domains correlates with MRTF-SRF activation. Following serum stimulation, nonpolymerizable actin dissociates from MRTF-A, and de novo formation of the G-actin-RPEL complex is impaired by a transferable factor. Our work demonstrates that WH2 domains activate MRTF-A and contribute to target gene regulation by a competitive mechanism, independently of their role in actin filament formation. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
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NASA Astrophysics Data System (ADS)
Oiwa, Nestor; Cordeiro, Claudette; Heermann, Dieter
2016-05-01
Instead of ATCG letter alignments, typically used in bioinformatics, we propose a new alignment method using the probability distribution function of the bottom of the occupied molecular orbital (BOMO), highest occupied molecular orbital (HOMO) and lowest unoccupied orbital (LUMO). We apply the technique to transcription factors with Cys2His2 zinc fingers. These transcription factors search for binding sites, probing for the electronic patterns at the minor and major DNA groves. The eukaryotic Cys2His2 zinc finger proteins bind to DNA ubiquitously at highly conserved domains. They are responsible for gene regulation and the spatial organization of DNA. To study and understand these zinc finger DNA-protein interactions, we use the extended ladder in the DNA model proposed by Zhu, Rasmussen, Balatsky & Bishop (2007) te{Zhu-2007}. Considering one single spinless electron in each nucleotide π-orbital along a double DNA chain (dDNA), we find a typical pattern for the bottom of BOMO, HOMO and LUMO along the binding sites. We specifically looked at two members of zinc finger protein family: specificity protein 1 (SP1) and early grown response 1 transcription factors (EGR1). When the valence band is filled, we find electrons in the purines along the nucleotide sequence, compatible with the electric charges of the binding amino acids in SP1 and EGR1 zinc finger.
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Terrados, Gloria; Finkernagel, Florian; Stielow, Bastian; Sadic, Dennis; Neubert, Juliane; Herdt, Olga; Krause, Michael; Scharfe, Maren; Jarek, Michael; Suske, Guntram
2012-01-01
The transcription factor Sp2 is essential for early mouse development and for proliferation of mouse embryonic fibroblasts in culture. Yet its mechanisms of action and its target genes are largely unknown. In this study, we have combined RNA interference, in vitro DNA binding, chromatin immunoprecipitation sequencing and global gene-expression profiling to investigate the role of Sp2 for cellular functions, to define target sites and to identify genes regulated by Sp2. We show that Sp2 is important for cellular proliferation that it binds to GC-boxes and occupies proximal promoters of genes essential for vital cellular processes including gene expression, replication, metabolism and signalling. Moreover, we identified important key target genes and cellular pathways that are directly regulated by Sp2. Most significantly, Sp2 binds and activates numerous sequence-specific transcription factor and co-activator genes, and represses the whole battery of cholesterol synthesis genes. Our results establish Sp2 as a sequence-specific regulator of vitally important genes. PMID:22684502
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Human HMG box transcription factor HBP1: a role in hCD2 LCR function.
Zhuma, T; Tyrrell, R; Sekkali, B; Skavdis, G; Saveliev, A; Tolaini, M; Roderick, K; Norton, T; Smerdon, S; Sedgwick, S; Festenstein, R; Kioussis, D
1999-01-01
The locus control region (LCR) of the human CD2 gene (hCD2) confers T cell-specific, copy-dependent and position-independent gene expression in transgenic mice. This LCR consists of a strong T cell-specific enhancer and an element without enhancer activity (designated HSS3), which is required for prevention of position effect variegation (PEV) in transgenic mice. Here, we identified the HMG box containing protein-1 (HBP1) as a factor binding to HSS3 of the hCD2 LCR. Within the LCR, HBP1 binds to a novel TTCATTCATTCA sequence that is higher in affinity than other recently reported HBP1-binding sites. Mice transgenic for a hCD2 LCR construct carrying a deletion of the HBP1-binding sequences show a propensity for PEV if the transgene integrates in a heterochromatic region of the chromosome such as the centromere or telomere. We propose that HBP1 plays an important role in chromatin opening and remodelling activities by binding to and bending the DNA, thus allowing DNA-protein and/or protein-protein interactions, which increase the probability of establishing an active locus. PMID:10562551
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Narayan, Vikram; Halada, Petr; Hernychová, Lenka; Chong, Yuh Ping; Žáková, Jitka; Hupp, Ted R.; Vojtesek, Borivoj; Ball, Kathryn L.
2011-01-01
The interferon-regulated transcription factor and tumor suppressor protein IRF-1 is predicted to be largely disordered outside of the DNA-binding domain. One of the advantages of intrinsically disordered protein domains is thought to be their ability to take part in multiple, specific but low affinity protein interactions; however, relatively few IRF-1-interacting proteins have been described. The recent identification of a functional binding interface for the E3-ubiquitin ligase CHIP within the major disordered domain of IRF-1 led us to ask whether this region might be employed more widely by regulators of IRF-1 function. Here we describe the use of peptide aptamer-based affinity chromatography coupled with mass spectrometry to define a multiprotein binding interface on IRF-1 (Mf2 domain; amino acids 106–140) and to identify Mf2-binding proteins from A375 cells. Based on their function as known transcriptional regulators, a selection of the Mf2 domain-binding proteins (NPM1, TRIM28, and YB-1) have been validated using in vitro and cell-based assays. Interestingly, although NPM1, TRIM28, and YB-1 all bind to the Mf2 domain, they have differing amino acid specificities, demonstrating the degree of combinatorial diversity and specificity available through linear interaction motifs. PMID:21245151
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HIV-1 Tat binds to SH3 domains: cellular and viral outcome of Tat/Grb2 interaction
Rom, Slava; Pacifici, Marco; Passiatore, Giovanni; Aprea, Susanna; Waligorska, Agnieszka; Valle, Luis Del; Peruzzi, Francesca
2011-01-01
The Src-homology 3 (SH3) domain is one of the most frequent protein recognition modules (PRMs), being represented in signal transduction pathways and in several pathologies such as cancer and AIDS. Grb2 (growth factor receptor-bound protein 2) is an adaptor protein that contains two SH3 domains and is involved in receptor tyrosine kinase (RTK) signal transduction pathways. The HIV-1 transactivator factor Tat is required for viral replication and it has been shown to bind directly or indirectly to several host proteins, deregulating their functions. In this study, we show interaction between the cellular factor Grb2 and the HIV-1 trans-activating protein Tat. The binding is mediated by the proline-rich sequence of Tat and the SH3 domain of Grb2. As the adaptor protein Grb2 participates in a wide variety of signaling pathways, we characterized at least one of the possible downstream effects of the Tat/Grb2 interaction on the well-known IGF-1R/Raf/MAPK cascade. We show that the binding of Tat to Grb2 impairs activation of the Raf/MAPK pathway, while potentiating the PKA/Raf inhibitory pathway. The Tat/Grb2 interaction affects also viral function by inhibiting the Tat-mediated transactivation of HIV-1 LTR and viral replication in infected primary microglia. PMID:21745501
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Trapnell, Cole; Davidson, Stuart; Pachter, Lior; Chu, Hou Cheng; Tonkin, Leath A.; Biggin, Mark D.; Eisen, Michael B.
2010-01-01
Changes in gene expression play an important role in evolution, yet the molecular mechanisms underlying regulatory evolution are poorly understood. Here we compare genome-wide binding of the six transcription factors that initiate segmentation along the anterior-posterior axis in embryos of two closely related species: Drosophila melanogaster and Drosophila yakuba. Where we observe binding by a factor in one species, we almost always observe binding by that factor to the orthologous sequence in the other species. Levels of binding, however, vary considerably. The magnitude and direction of the interspecies differences in binding levels of all six factors are strongly correlated, suggesting a role for chromatin or other factor-independent forces in mediating the divergence of transcription factor binding. Nonetheless, factor-specific quantitative variation in binding is common, and we show that it is driven to a large extent by the gain and loss of cognate recognition sequences for the given factor. We find only a weak correlation between binding variation and regulatory function. These data provide the first genome-wide picture of how modest levels of sequence divergence between highly morphologically similar species affect a system of coordinately acting transcription factors during animal development, and highlight the dominant role of quantitative variation in transcription factor binding over short evolutionary distances. PMID:20351773
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Fjellström, Ola; Akkaya, Sibel; Beisel, Hans-Georg; Eriksson, Per-Olof; Erixon, Karl; Gustafsson, David; Jurva, Ulrik; Kang, Daiwu; Karis, David; Knecht, Wolfgang; Nerme, Viveca; Nilsson, Ingemar; Olsson, Thomas; Redzic, Alma; Roth, Robert; Sandmark, Jenny; Tigerström, Anna; Öster, Linda
2015-01-01
Activated factor XI (FXIa) inhibitors are anticipated to combine anticoagulant and profibrinolytic effects with a low bleeding risk. This motivated a structure aided fragment based lead generation campaign to create novel FXIa inhibitor leads. A virtual screen, based on docking experiments, was performed to generate a FXIa targeted fragment library for an NMR screen that resulted in the identification of fragments binding in the FXIa S1 binding pocket. The neutral 6-chloro-3,4-dihydro-1H-quinolin-2-one and the weakly basic quinolin-2-amine structures are novel FXIa P1 fragments. The expansion of these fragments towards the FXIa prime side binding sites was aided by solving the X-ray structures of reported FXIa inhibitors that we found to bind in the S1-S1’-S2’ FXIa binding pockets. Combining the X-ray structure information from the identified S1 binding 6-chloro-3,4-dihydro-1H-quinolin-2-one fragment and the S1-S1’-S2’ binding reference compounds enabled structure guided linking and expansion work to achieve one of the most potent and selective FXIa inhibitors reported to date, compound 13, with a FXIa IC50 of 1.0 nM. The hydrophilicity and large polar surface area of the potent S1-S1’-S2’ binding FXIa inhibitors compromised permeability. Initial work to expand the 6-chloro-3,4-dihydro-1H-quinolin-2-one fragment towards the prime side to yield molecules with less hydrophilicity shows promise to afford potent, selective and orally bioavailable compounds. PMID:25629509
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Nelson, Christopher S; Fuller, Chris K; Fordyce, Polly M; Greninger, Alexander L; Li, Hao; DeRisi, Joseph L
2013-07-01
The transcription factor forkhead box P2 (FOXP2) is believed to be important in the evolution of human speech. A mutation in its DNA-binding domain causes severe speech impairment. Humans have acquired two coding changes relative to the conserved mammalian sequence. Despite intense interest in FOXP2, it has remained an open question whether the human protein's DNA-binding specificity and chromatin localization are conserved. Previous in vitro and ChIP-chip studies have provided conflicting consensus sequences for the FOXP2-binding site. Using MITOMI 2.0 microfluidic affinity assays, we describe the binding site of FOXP2 and its affinity profile in base-specific detail for all substitutions of the strongest binding site. We find that human and chimp FOXP2 have similar binding sites that are distinct from previously suggested consensus binding sites. Additionally, through analysis of FOXP2 ChIP-seq data from cultured neurons, we find strong overrepresentation of a motif that matches our in vitro results and identifies a set of genes with FOXP2 binding sites. The FOXP2-binding sites tend to be conserved, yet we identified 38 instances of evolutionarily novel sites in humans. Combined, these data present a comprehensive portrait of FOXP2's-binding properties and imply that although its sequence specificity has been conserved, some of its genomic binding sites are newly evolved.
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Nelson, Christopher S.; Fuller, Chris K.; Fordyce, Polly M.; Greninger, Alexander L.; Li, Hao; DeRisi, Joseph L.
2013-01-01
The transcription factor forkhead box P2 (FOXP2) is believed to be important in the evolution of human speech. A mutation in its DNA-binding domain causes severe speech impairment. Humans have acquired two coding changes relative to the conserved mammalian sequence. Despite intense interest in FOXP2, it has remained an open question whether the human protein’s DNA-binding specificity and chromatin localization are conserved. Previous in vitro and ChIP-chip studies have provided conflicting consensus sequences for the FOXP2-binding site. Using MITOMI 2.0 microfluidic affinity assays, we describe the binding site of FOXP2 and its affinity profile in base-specific detail for all substitutions of the strongest binding site. We find that human and chimp FOXP2 have similar binding sites that are distinct from previously suggested consensus binding sites. Additionally, through analysis of FOXP2 ChIP-seq data from cultured neurons, we find strong overrepresentation of a motif that matches our in vitro results and identifies a set of genes with FOXP2 binding sites. The FOXP2-binding sites tend to be conserved, yet we identified 38 instances of evolutionarily novel sites in humans. Combined, these data present a comprehensive portrait of FOXP2’s-binding properties and imply that although its sequence specificity has been conserved, some of its genomic binding sites are newly evolved. PMID:23625967
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Synthetic heparin-binding growth factor analogs
Pena, Louis A.; Zamora, Paul; Lin, Xinhua; Glass, John D.
2007-01-23
The invention provides synthetic heparin-binding growth factor analogs having at least one peptide chain that binds a heparin-binding growth factor receptor, covalently bound to a hydrophobic linker, which is in turn covalently bound to a non-signaling peptide that includes a heparin-binding domain. The synthetic heparin-binding growth factor analogs are useful as soluble biologics or as surface coatings for medical devices.
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Madabhushi, Sri R; Shang, Chengwei; Dayananda, Kannayakanahalli M; Rittenhouse-Olson, Kate; Murphy, Mary; Ryan, Thomas E; Montgomery, Robert R; Neelamegham, Sriram
2012-05-17
Noncovalent association between the von Willebrand factor (VWF) propeptide (VWFpp) and mature VWF aids N-terminal multimerization and protein compartmentalization in storage granules. This association is currently thought to dissipate after secretion into blood. In the present study, we examined this proposition by quantifying the affinity and kinetics of VWFpp binding to mature VWF using surface plasmon resonance and by developing novel anti-VWF D'D3 mAbs. Our results show that the only binding site for VWFpp in mature VWF is in its D'D3 domain. At pH 6.2 and 10mM Ca(2+), conditions mimicking intracellular compartments, VWFpp-VWF binding occurs with high affinity (K(D) = 0.2nM, k(off) = 8 × 10(-5) s(-1)). Significant, albeit weaker, binding (K(D) = 25nM, k(off) = 4 × 10(-3) s(-1)) occurs under physiologic conditions of pH 7.4 and 2.5mM Ca(2+). This interaction was also observed in human plasma (K(D) = 50nM). The addition of recombinant VWFpp in both flow-chamber-based platelet adhesion assays and viscometer-based shear-induced platelet aggregation and activation studies reduced platelet adhesion and activation partially. Anti-D'D3 mAb DD3.1, which blocks VWFpp binding to VWF-D'D3, also abrogated platelet adhesion, as shown by shear-induced platelet aggregation and activation studies. Our data demonstrate that VWFpp binding to mature VWF occurs in the circulation, which can regulate the hemostatic potential of VWF by reducing VWF binding to platelet GpIbα.
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Rodriguez, C; Huang, L J; Son, J K; McKee, A; Xiao, Z; Lodish, H F
2001-08-10
Using the plasminogen activator inhibitor (PAI) promoter to drive the expression of a reporter gene (mouse CD2), we devised a system to clone negative regulators of the transforming growth factor-beta (TGF-beta) signaling pathway. We infected a TGF-beta-responsive cell line (MvLu1) with a retroviral cDNA library, selecting by fluorescence-activated cell sorter single cells displaying low PAI promoter activity in response to TGF-beta. Using this strategy we cloned the proto-oncogene brain factor-1 (BF-1). BF-1 represses the PAI promoter in part by associating with both unphosphorylated Smad3 (in the cytoplasm) and phosphorylated Smad3 (in the nucleus), thus preventing its binding to DNA. BF-1 also associates with Smad1, -2, and -4; the Smad MH2 domain binds to BF-1, and the C-terminal segment of BF-1 is uniquely and solely required for binding to Smads. Further, BF-1 represses another TGF-beta-induced promoter (p15), it up-regulates a TGF-beta-repressed promoter (Cyclin A), and it reverses the growth arrest caused by TGF-beta. Our results suggest that BF-1 is a general inhibitor of TGF-beta signaling and as such may play a key role during brain development.
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Structure and Biophysics of CBFβ/RUNX and Its Translocation Products.
Tahirov, Tahir H; Bushweller, John
2017-01-01
The core binding factor (CBF) transcription factor is somewhat unique in that it is composed of a DNA binding RUNX subunit (RUNX1, 2, or 3) and a non-DNA binding CBFβ subunit, which modulates RUNX protein activity by modulating the auto-inhibition of the RUNX subunits. Since the discovery of this fascinating transcription factor more than 20 years ago, there has been a robust effort to characterize the structure as well as the biochemical properties of CBF. More recently, these efforts have also extended to the fusion proteins that arise from the subunits of CBF in leukemia. This chapter highlights the work of numerous labs which has provided a detailed understanding of the structure and function of this transcription factor and its fusion proteins.
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Shashoua, V E; Adams, D; Boyer-Boiteau, A
2001-10-19
An 8-amino acid peptide fragment (CMX-8933) of Ependymin, a glycoprotein component of the extracellular fluid and cerebrospinal fluid of goldfish brain, was synthesized and tested for its capacity to activate AP-1 transcription factor in cell cultures. Dose-response and time-course studies of AP-1's binding to DNA were carried out in neuroblastoma (NB2a/dl) and primary rat brain cortical cultures using an electrophoretic mobility shift assay (EMSA). A 13-14-fold increase in AP-1's DNA binding was obtained when NB2a cells were incubated for 4 h with 6-10 microg/ml CMX-8933. Primary rat brain cortical cultures were much more sensitive to the effects of CMX-8933 than transformed (NB2a) cultures; here a 26.7+/-5.2-fold increase in binding was observed following a 3-h treatment with as little as 10 ng/ml peptide. These findings are consistent with an activation of this transcription factor, a characteristic that has been previously correlated with functional aspects of full-sized neurotrophic factors (nerve growth factor and brain-derived nerve growth factor) in neuronal differentiation and regeneration. Such data suggest a role for Ependymin in transcriptional control.
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Schiering, Nikolaus; Knapp, Stefan; Marconi, Marina; Flocco, Maria M; Cui, Jean; Perego, Rita; Rusconi, Luisa; Cristiani, Cinzia
2003-10-28
The protooncogene c-met codes for the hepatocyte growth factor receptor tyrosine kinase. Binding of its ligand, hepatocyte growth factor/scatter factor, stimulates receptor autophosphorylation, which leads to pleiotropic downstream signaling events in epithelial cells, including cell growth, motility, and invasion. These events are mediated by interaction of cytoplasmic effectors, generally through Src homology 2 (SH2) domains, with two phosphotyrosine-containing sequence motifs in the unique C-terminal tail of c-Met (supersite). There is a strong link between aberrant c-Met activity and oncogenesis, which makes this kinase an important cancer drug target. The furanosylated indolocarbazole K-252a belongs to a family of microbial alkaloids that also includes staurosporine. It was recently shown to be a potent inhibitor of c-Met. Here we report the crystal structures of an unphosphorylated c-Met kinase domain harboring a human cancer mutation and its complex with K-252a at 1.8-A resolution. The structure follows the well established architecture of protein kinases. It adopts a unique, inhibitory conformation of the activation loop, a catalytically noncompetent orientation of helix alphaC, and reveals the complete C-terminal docking site. The first SH2-binding motif (1349YVHV) adopts an extended conformation, whereas the second motif (1356YVNV), a binding site for Grb2-SH2, folds as a type II Beta-turn. The intermediate portion of the supersite (1353NATY) assumes a type I Beta-turn conformation as in an Shc-phosphotyrosine binding domain peptide complex. K-252a is bound in the adenosine pocket with an analogous binding mode to those observed in previously reported structures of protein kinases in complex with staurosporine.
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Structure, dynamics and RNA binding of the multi-domain splicing factor TIA-1
Wang, Iren; Hennig, Janosch; Jagtap, Pravin Kumar Ankush; Sonntag, Miriam; Valcárcel, Juan; Sattler, Michael
2014-01-01
Alternative pre-messenger ribonucleic acid (pre-mRNA) splicing is an essential process in eukaryotic gene regulation. The T-cell intracellular antigen-1 (TIA-1) is an apoptosis-promoting factor that modulates alternative splicing of transcripts, including the pre-mRNA encoding the membrane receptor Fas. TIA-1 is a multi-domain ribonucleic acid (RNA) binding protein that recognizes poly-uridine tract RNA sequences to facilitate 5′ splice site recognition by the U1 small nuclear ribonucleoprotein (snRNP). Here, we characterize the RNA interaction and conformational dynamics of TIA-1 by nuclear magnetic resonance (NMR), isothermal titration calorimetry (ITC) and small angle X-ray scattering (SAXS). Our NMR-derived solution structure of TIA-1 RRM2–RRM3 (RRM2,3) reveals that RRM2 adopts a canonical RNA recognition motif (RRM) fold, while RRM3 is preceded by an non-canonical helix α0. NMR and SAXS data show that all three RRMs are largely independent structural modules in the absence of RNA, while RNA binding induces a compact arrangement. RRM2,3 binds to pyrimidine-rich FAS pre-mRNA or poly-uridine (U9) RNA with nanomolar affinities. RRM1 has little intrinsic RNA binding affinity and does not strongly contribute to RNA binding in the context of RRM1,2,3. Our data unravel the role of binding avidity and the contributions of the TIA-1 RRMs for recognition of pyrimidine-rich RNAs. PMID:24682828
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Stepanchick, Ann; Zhi, Huijun; Cavanaugh, Alice H; Rothblum, Katrina; Schneider, David A; Rothblum, Lawrence I
2013-03-29
The human homologue of yeast Rrn3 is an RNA polymerase I-associated transcription factor that is essential for ribosomal DNA (rDNA) transcription. The generally accepted model is that Rrn3 functions as a bridge between RNA polymerase I and the transcription factors bound to the committed template. In this model Rrn3 would mediate an interaction between the mammalian Rrn3-polymerase I complex and SL1, the rDNA transcription factor that binds to the core promoter element of the rDNA. In the course of studying the role of Rrn3 in recruitment, we found that Rrn3 was in fact a DNA-binding protein. Analysis of the sequence of Rrn3 identified a domain with sequence similarity to the DNA binding domain of heat shock transcription factor 2. Randomization, or deletion, of the amino acids in this region in Rrn3, amino acids 382-400, abrogated its ability to bind DNA, indicating that this domain was an important contributor to DNA binding by Rrn3. Control experiments demonstrated that these mutant Rrn3 constructs were capable of interacting with both rpa43 and SL1, two other activities demonstrated to be essential for Rrn3 function. However, neither of these Rrn3 mutants was capable of functioning in transcription in vitro. Moreover, although wild-type human Rrn3 complemented a yeast rrn3-ts mutant, the DNA-binding site mutant did not. These results demonstrate that DNA binding by Rrn3 is essential for transcription by RNA polymerase I.