YY1 functions in vascular smooth muscle cells

YY1在血管平滑肌细胞中发挥作用

基本信息

批准号：
7371940
负责人：
ANNY A USHEVA-SIMIDJIYSKA
金额：
$ 32.24万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7371940
关键词：
Acetylglucosamine Affinity Amino Acids Binding Blood Vessels Carotid Arteries Cell Extracts Cell Fraction Cell Proliferation Clinical Medicine Cultured Cells Flow Cytometry Gene Expression Gene Expression Regulation Genetic Transcription Goals Growth In Vitro Injury Ions Mass Spectrum Analysis Modeling Monosaccharides Numbers Phosphorylation Positioning Attribute Post-Translational Protein Processing Primary Cell Cultures Proliferating Protein Glycosylation Protein Overexpression Proteins Proteomics Rate Rattus Regulation Reporter Research Personnel Serine Site Site-Directed Mutagenesis Smooth Muscle Smooth Muscle Myocytes Structure Surface Testing Threonine Transfection Variant Viral base chromatin immunoprecipitation day expression vector glycosylation in vivo injured interest mutant programs promoter research study response response to injury tool

项目摘要

DESCRIPTION (provided by applicant): Activation of smooth muscle cell (SMC) proliferation in response to vascular injury is a widely appreciated vascular problem in clinical medicine, but its basis remains obscure. Our hypothesis is that abnormal posttranslational modification of proteins makes a major contribution to the initiation of activated smooth muscle cellular proliferation associated with vascular neointimal formation following vascular injury. We are particularly interested in the possibility that O-GlcNAcylation of YY1, significantly elevated in the neontimal smooth muscle cells, regulates the YY1-Rb interaction and downstream transcription promoting gene expression that triggers smooth muscle cellular proliferation. We are interested in the possibility that the phosphorylation of Rb at specific amino acids also inhibits the YY1-Rb complexation contributing to the activation of smooth muscle cellular proliferation. The goal of this project is to identify how O-glycosylation of YY1 regulates the YY1-Rb interaction and the transcriptional changes that occur in SMC transition from quiescence to active proliferation. Aim 1: Determine the specific sites of YY1 O-GlcNAcylation in growth arrested and in growth stimulated SMC cultures. The position of O-GlcNAc attachments to YY1 in the fractions will be determined by applying a combination of automated triple quadruple and ion trapping tandem mass spectroscopy. Aim 2: Analysis of YY1 O-GlcNAcylation in response to vascular balloon injury in a rat carotid artery model. The rate and the position of YY1 O-glycosylation 2 days, 4 days, and 8 days after rat balloon injury will be determined. Aim 3: Structure-based site-directed mutagenesis for the identification of the YY1-Rb binding surface and its regulation by Rb phosphorylation. The phosphorylated Ser/Thr residues on pRb that inhibit YY1 binding will be identified. Aim 4: Determine how O-GlcNAcylation influences cellular YY1 functions applying flow cytometry, gene expression regulation, and chromatin immunoprecipitation (ChIP).

描述（由申请人提供）：响应血管损伤的平滑肌细胞（SMC）增殖的激活是临床医学中广泛理解的血管问题，但其基础仍然晦涩难懂。我们的假设是，蛋白质的异常翻译后修饰对血管损伤后血管新内膜形成相关的活化平滑肌细胞增殖的启动作出了重大贡献。我们对YY1的O-glcnacylation的可能性特别感兴趣，在新生儿平滑肌细胞中显着升高，调节YY1-RB的相互作用和下游转录，从而促进了基因表达，从而促进了平滑肌细胞增殖。我们对特异性氨基酸的RB磷酸化也抑制了有助于平滑肌细胞增殖的激活的YY1-RB络合物的可能性感兴趣。该项目的目的是确定YY1的O-糖基化如何调节YY1-RB相互作用以及SMC从静止增殖到主动增殖的转录变化。 AIM 1：确定YY1 O-Glcnacylation在被捕的生长和生长刺激的SMC培养中的特定部位。 O-GlCNAC附着在YY1中的位置将通过应用自动三倍四倍体和离子捕获串联质谱的组合来确定。 AIM 2：在大鼠颈动脉模型中响应血管球囊损伤的YY1 O-Glcnacylation分析。 YY1 O-糖基化的速率和位置将在大鼠球囊损伤后2天，4天和8天确定。 AIM 3：基于结构的位置定向诱变，用于鉴定YY1-RB结合表面及其通过RB磷酸化调节。将鉴定出抑制YY1结合的PRB上的磷酸化Ser/ThR残基。目标4：确定O-Glcnacylation如何影响细胞YY1的功能，该功能应用流式细胞仪，基因表达调节和染色质免疫沉淀（CHIP）。

项目成果

期刊论文数量（21）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

DNA breathing dynamics distinguish binding from nonbinding consensus sites for transcription factor YY1 in cells.

DOI：
10.1093/nar/gks758
发表时间：
2012-11-01
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Alexandrov BS;Fukuyo Y;Lange M;Horikoshi N;Gelev V;Rasmussen KØ;Bishop AR;Usheva A
通讯作者：
Usheva A

Oxidative stress plays a critical role in inactivating mutant BRAF by geldanamycin derivatives.

DOI：
10.1158/0008-5472.can-07-6602
发表时间：
2008-08-01
期刊：
Cancer research
影响因子：
11.2
作者：
Fukuyo Y;Inoue M;Nakajima T;Higashikubo R;Horikoshi NT;Hunt C;Usheva A;Freeman ML;Horikoshi N
通讯作者：
Horikoshi N

YY1-DNA interaction results in a significant change of electronic context as measured by capacitance.

YY1-DNA 相互作用导致通过电容测量的电子环境发生显着变化。

DOI：
10.1016/s0301-4622(02)00236-3
发表时间：
2003
期刊：
Biophysical chemistry
影响因子：
3.8
作者：
Choi,ChuH;Moser,Joel;Sabourin,NicaulasA;Blagoev,Krastan;Naughton,Michael;Usheva,Anny;Sobourin,Nicaulas
通讯作者：
Sobourin,Nicaulas

Modulating the dysregulated migration of pulmonary arterial hypertensive smooth muscle cells with motif mimicking cell permeable peptides.

用基序模拟细胞通透性肽调节肺动脉高血压平滑肌细胞的失调迁移。

DOI：
发表时间：
2015
期刊：
Current topics in peptide & protein research
影响因子：
0
作者：
Wilson,JamieL;Rupasinghe,Chamila;Usheva,Anny;Warburton,Rod;Kaplan,Chloe;Taylor,Linda;Hill,Nicholas;Mierke,DaleF;Polgar,Peter
通讯作者：
Polgar,Peter

DNA dynamics is likely to be a factor in the genomic nucleotide repeats expansions related to diseases.

DOI：
10.1371/journal.pone.0019800
发表时间：
2011
期刊：
PloS one
影响因子：
3.7
作者：
Alexandrov BS;Valtchinov VI;Alexandrov LB;Gelev V;Dagon Y;Bock J;Kohane IS;Rasmussen KØ;Bishop AR;Usheva A
通讯作者：
Usheva A