UV regulation of anti-apoptotic co-repressor CtBP

抗凋亡辅阻遏物 CtBP 的紫外线调节

基本信息

批准号：
8079122
负责人：
QINGHONG ZHANG
金额：
$ 28.2万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8079122
关键词：
Affinity Chromatography Antibodies Apoptosis Apoptotic Binding Proteins Biochemical Biological Assay Cancer Biology Cell Adhesion Molecules Cell Death Cellular Stress Cervix carcinoma Chromatin Code Complex DNA Binding Data Down-Regulation Embryo Epithelial Fibroblasts Functional RNA Gene Targeting Genes Genetic Transcription Hela Cells Human JUN gene Knock-out Knockout Mice Lead Malignant Neoplasms Mass Spectrum Analysis Mediating MicroRNAs Modification Molecular Mutation Pathway interactions Peptides Phospho-Specific Antibodies Phosphorylation Phosphotransferases Polyubiquitination Proteins Regulation Research Personnel Role Signal Pathway Signal Transduction Signal Transduction Pathway Small Interfering RNA Stimulus System Testing Tumor Suppressor Genes Tumor Suppressor Proteins Ubiquitination base cancer therapy chromatin immunoprecipitation design in vivo insight lead-binding proteins mutant novel novel strategies programs protein degradation research study response stress-activated protein kinase 1 tumor growth tumorigenesis ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): CtBP is a transcriptional co-repressor for a panel of pro-apoptotic genes. Its degradation leads to cell death independent of p53. Furthermore, knockdown of CtBP is sufficient to suppress tumorigenesis in vivo. This proposal is designed to study the mechanisms of CtBP degradation in the hope of developing novel approaches for cancer treatment. The specific aims are to: 1) Determine whether HIPK2 regulates CtBP directly or through other kinase pathways. Phosphorylation of Ser 422 triggers CtBP for degradation. We have raised an antibody to Ser 422-phosphorylated CtBP to examine whether HIPK2 interacts with other signal pathways to phosphorylate CtBP and trigger CtBP degradation. JNK1 specifically phosphorylates Ser 422. We will investigate the involvement of this pathway in CtBP degradation. 2) Determine how phosphorylation of CtBP leads to proteasomal degradation. We will focus on the identification of E3 ligase(s) responsible for CtBP polyubiquitination and degradation. The tumor suppressor Fbw7a physically interacts with CtBP and is our first candidate. We have shown that a Ser 422 phospho-peptide blocks CtBP degradation. We will use affinity chromatography and mass spectrometry to identify proteins associated with this phospho-peptide. The interaction of the targeting proteins with the phospho-peptide will be confirmed by biochemical assays and their contribution to CtBP degradation will be examined using siRNA. 3) Identify CtBP target genes involved in transformation and apoptosis. CtBP represses many epithelial- specific and pro-apoptotic genes. Whether the genes responsible for these effects are direct CtBP targets is uncertain. We will identify CtBP targets (both protein-coding and non-coding such as microRNAs) involved in apoptosis arid transformation using Serial Analysis of Chromatin Occupancy, which combines chromatin immunoprecipitation with long SAGE. Expression of target genes involved in apoptosis and transformation will be studied in specific signaling pathways that cause CtBP degradation.

描述（由申请人提供）：CTBP是促凋亡基因小组的转录共抑制剂。它的降解导致细胞死亡与p53无关。此外，CTBP的敲低足以抑制体内肿瘤发生。该建议旨在研究CTBP降解的机制，以期开发新的癌症治疗方法。具体目的是：1）确定HIPK2是直接调节CTBP还是通过其他激酶途径调节CTBP。 Ser 422触发CTBP降解的磷酸化。我们已经提出了Ser 422-磷酸化CTBP的抗体，以检查HIPK2是否与其他信号途径相互作用，以磷酸化CTBP并触发CTBP降解。 JNK1特异性地磷酸化了Ser 422。我们将研究该途径在CTBP降解中的参与。 2）确定CTBP的磷酸化如何导致蛋白酶体降解。我们将重点关注负责CTBP多泛素化和降解的E3连接酶的鉴定。肿瘤抑制剂FBW7A与CTBP物理相互作用，是我们的第一个候选者。我们已经表明，Ser 422磷酸肽会阻止CTBP降解。我们将使用亲和色谱和质谱法来鉴定与该磷酸肽相关的蛋白质。靶蛋白与磷酸肽的相互作用将通过生化测定确认，并将使用siRNA检查其对CTBP降解的贡献。 3）确定与转化和凋亡有关的CTBP靶基因。 CTBP抑制许多上皮特异性和促凋亡基因。负责这些效应的基因是否是直接的CTBP靶标。我们将使用染色质占用率的系列分析来鉴定参与凋亡干旱转化的CTBP靶标（蛋白质编码和非编码），该靶标将染色质免疫沉淀与长SAGE结合在一起。将在导致CTBP降解的特定信号通路中研究参与凋亡和转化的靶基因的表达。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Display of cell surface sites for fibronectin assembly is modulated by cell adherence to (1)F3 and C-terminal modules of fibronectin.

纤连蛋白组装的细胞表面位点的显示是通过细胞粘附到纤连蛋白的 (1)F3 和 C 末端模块来调节的。

DOI：
10.1371/journal.pone.0004113
发表时间：
2009
期刊：
PloS one
影响因子：
3.7
作者：
Xu,Jielin;Bae,Eunnyung;Zhang,Qinghong;Annis,DouglasS;Erickson,HaroldP;Mosher,DeaneF
通讯作者：
Mosher,DeaneF

MicroRNA-137 targets carboxyl-terminal binding protein 1 in melanoma cell lines.