Proteins encoded by oncogenes such as v-fps/fes, v-src, v-yes, v-abl, and v-fgr are cytoplasmic protein tyrosine kinases which, unlike transmembrane receptors, localized to the inside of cell. These proteins possess two contiguous regions sequence identity: a C-terminal catalytic domain 260 residues with homology other tyrosine-specific serine-threonine-specific kinases, unique approximately 100 which is located N terminal kinase region absent from that span plasma membrane. In-frame linker...

Proteins encoded by oncogenes such as v-fps/fes, v-src, v-yes, v-abl, and v-fgr are cytoplasmic protein tyrosine kinases which, unlike transmembrane receptors, localized to the inside of cell. These proteins possess two contiguous regions sequence identity: a C-terminal catalytic domain 260 residues with homology other tyrosine-specific serine-threonine-specific kinases, unique approximately 100 which is located N terminal kinase region absent from that span plasma membrane. In-frame linker...

PhosphoGRID is an online database that curates and houses experimentally verified in vivo phosphorylation sites the Saccharomyces cerevisiae proteome (www.phosphogrid.org). Phosphosites are annotated with specific protein kinases and/or phosphatases, along condition(s) under which occurs effects on function. We report here updated data set, including nine additional high-throughput (HTP) mass spectrometry studies. The version 2.0 set contains information 20 177 unique phosphorylated...

Protein phosphorylation plays a central role in cellular regulation. Recent proteomics strategies for identifying phosphopeptides have been developed using the model organism Saccharomyces cerevisiae, and consequently, when combined with studies of individual gene products, number reported specific sites this has expanded enormously. In order to systematically document integrate these various data types, we database experimentally verified vivo curated from S. cerevisiae primary literature....

Hortaea werneckii, ascomycetous yeast from the order Capnodiales, shows an exceptional adaptability to osmotically stressful conditions. To investigate this unusual phenotype we obtained a draft genomic sequence of H. werneckii strain isolated hypersaline water solar saltern. Two its most striking characteristics that may be associated with halotolerant lifestyle are large genetic redundancy and expansion genes encoding metal cation transporters. Although no sexual state has yet been...

HIV-1 latency poses a major barrier to viral eradication. Canonically, is thought arise from progressive epigenetic silencing of active infections. However, little known about when and how long terminal repeat (LTR)-silent infections since the majority current models cannot differentiate between initial (LTR-silent) secondary (progressive silencing) latency. In this study, we constructed characterized novel, double-labeled vector (Red-Green-HIV-1 [RGH]) that allows for detection infected...

Latent HIV-1 (human immunodeficiency virus-1) provirus is unaffected by current AIDS (acquired syndrome) therapies. We show here that chaetocin, an SUV39H1 histone methyltransferase inhibitor, causes 25-fold induction of latent expression, while producing minimal toxicity and without causing T cell activation. Induction associated with loss H3 lysine 9 (H3K9) trimethylation at the long terminal repeat (LTR) promoter, a corresponding increase in H3K9 acetylation. The effect chaetocin...

The ability to determine the global location of transcription factor binding sites in vivo is important for a comprehensive understanding gene regulation human cells. We have developed technology, called serial analysis elements (SABE), involving subtractive hybridization chromatin immunoprecipitation-enriched DNA fragments followed by generation and concatamerized sequence tags. applied SABE technology search p53 target genes genome, identified several previously described targets addition...

A conserved noncatalytic domain SH2 (for src homology region 2) is located immediately N terminal to the kinase domains of all cytoplasmic protein-tyrosine kinases. We found that wild-type v-fps stimulated enzymatic activity adjacent 10-fold and functioned as a powerful positive effector catalytic transforming activities within oncoprotein (P130gag-fps). Partial proteolysis P130gag-fps supporting genetic data indicated exerts its effect on through an intramolecular interaction with domain....

All cytoplasmic protein-tyrosine kinases (PTKs) share a noncatalytic domain, termed SH2, which comprises approximately 100 residues located immediately N-terminal to the kinase domain. A linker in AX9m mutant of Fujinami avian sarcoma virus (FSV) introduces dipeptide insertion into SH2 domain P130gag-fps PTK, abolishes its ability transform Rat-2 cells. However, at 36 degrees C FSV elicits focus formation and agar colony infected chicken embryo fibroblasts (CEF) with single hit kinetics. At...

Binding of USF1/2 and TFII-I (RBF-2) at conserved sites flanking the HIV-1 LTR enhancer is essential for reactivation from latency in T cells, with knockdown rendering provirus insensitive to cell signaling. We identified an interaction tripartite motif protein TRIM24, these factors were found be constitutively associated LTR. Similar effect depletion, loss TRIM24 impaired response deficiency did not affect recruitment RNA Pol II promoter, but inhibited transcriptional elongation, that was...

AbstractThe yeast Saccharomyces cerevisiae transcription factor Ste12p is responsible for activating genes in response to MAP kinase cascades controlling mating and filamentous growth. negatively regulated by two inhibitor proteins, Dig1p (also called Rst1p) Dig2p Rst2p). The expression of a C-terminal fragment (residues 216 688) [Ste12p(216–688)] from aGAL promoter causes FUS1 induction strain expressing wild-type STE12, suggesting that this region can cause the activation endogenous...

Despite extensive in vitro studies identifying a myriad of cellular transcription factors that bind the human immunodeficiency virus type 1 5' long terminal repeat (LTR), relative contribution these to replication infected individuals remains obscure. To address this question, we investigated 478 proviral quasispecies derived from uncultured peripheral blood mononuclear cells 42 patients representing all stages infection. In addition highly conserved TATA box, SP-1, and NF-kappaB sites, Ets...

Activating region I of GAL4 protein is a stretch amino acids, positioned adjacent to the DNA-binding region, that activates transcription in yeast and, as we show here, mammalian cells. Here describe mutations located throughout 65-amino acid increase activation function I. Most these replace positively charged acids with neutral ones, although also substitutions at one position do not alter charge region. Mutations have similar effects on when assayed When individual raise acidity...

Abstract Background Molecular latency allows HIV-1 to persist in resting memory CD4+ T-cells as transcriptionally silent provirus integrated into host chromosomal DNA. Multiple transcriptional regulatory mechanisms for have been described the context of progressive epigenetic silencing and maintenance. However, our understanding determinants critical establishment newly infected cells is limited. Results In this study, we used a recently described, doubly fluorescent model dissect role...

Expression of the HIV-1 genome by RNA Polymerase II is regulated at multiple steps, as are most cellular genes, including recruitment general transcription factors and control transcriptional elongation from core promoter. We recently discovered that tripartite motif protein TRIM24 recruited to Long Terminal Repeat (LTR) interaction with TFII-I causes stimulating association PTEF-b/ CDK9. Because required for stimulation LTR, we were surprised find IACS-9571, a specific inhibitor C-terminal...

Latent HIV-1 provirus represents the barrier toward a cure for infection and is dependent upon host RNA Polymerase (Pol) II machinery reemergence. Here, we find that inhibitors of Pol mediator kinases CDK8/19, Senexin A BRD6989, inhibit induction expression in response to latency-reversing agents T cell signaling agonists. These were found impair recruitment LTR. Furthermore, several latency reversal was impaired disruption CDK8 by shRNA or gene knockout. However, effects depletion did not...

Gal4p activates transcription of the Saccharomyces GALgenes in response to galactose and is phosphorylated during interaction with RNA polymerase II (Pol II) holoenzyme. One phosphorylation at S699 necessary for full GAL induction mediated by Srb10p/CDK8 Pol holoenzyme mediator subcomplex. sensitive inducer, its requirement can be abrogated high concentrations strains expressing wild-typeGAL2 GAL3. occurs independently Gal3p responsible long-term adaptation observed gal3 yeast.SRB10 GAL3 are...