Function of RVB1-Tip60 in the DNA damage response

RVB1-Tip60 在 DNA 损伤反应中的功能

基本信息

批准号：
7581450
负责人：
Anindya Dutta
金额：
$ 29.54万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-26 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7581450
关键词：
ATP phosphohydrolase ATPase Domain Acetylation Acetyltransferase Active Sites Arginine Attention Binding Binding Proteins Biology Cancer Etiology Cell physiology Cells Chemicals Chromatin Chromatin Remodeling Factor Complex DNA DNA Damage DNA Double Strand Break Down-Regulation Fingers Gene Expression Genes H2AFX gene HTATIP gene Histone Acetylation Histone H4 Histones Homologous Gene Human Hydrolysis Laboratories Lead Malignant Neoplasms Measures Metabolism Mitomycin Mitomycins Molecular Molecular Profiling Mutate Mutation Nucleosomes Phenotype Phosphoric Monoester Hydrolases Phosphorylation Play Protein Dephosphorylation Proteins Public Health Publishing Radiation therapy Radio Recruitment Activity Regulation Relative (related person)Role Site Structure Testing Therapeutic Transcriptional Activation Transferase Up-Regulation Variant Walkers Yeasts base cancer cell cancer therapy chemotherapy chromatin remodeling cisplatin/etoposide protocol design histone acetyltransferase human H2AX protein human HTATIP protein in vitro Assay in vivo inhibitor/antagonist programs promoter protein function response

项目摘要

DESCRIPTION (provided by applicant): Two important molecular signatures of a human cell's response to DNA damage are the phosphorylation of a histone variant H2AX and a change in the transcriptional program. Recent results from our laboratory have implicated the RVB1 protein in the cellular response to DNA damage. Human RVB is an integral part of a chromatin remodeling complex containing the histone acetyltransferase, TIP60, the ATPase p400 and several other cellular proteins. RVB1 or TIP60 depletion leads to an increase in phosphoH2AX accumulation after DNA damage and sensitizes cancer cells to DNA damaging agents, suggesting that inhibitors of RVB ATPase may be useful in chemo- or radiotherapy. This project will test two hypotheses: 1) RVB1 is required to neutralize the inhibitory action of p400 on the acetyltransferase activity of the TIP60 complex and 2) the acetyltransferase activity of TIP60 is required to mobilize phosphoH2AX containing nucleosomes off the DNA prior to its dephosphorylation by cellular phosphatases. Aim 1 will explore whether TIP60 is the primary chromatin remodeler involved in down modulating phosphoH2AX and whether RVB2 co-operates with RVB1 in this function. Aim 2 will test whether the hypothesis of TIP60 regulation by RVB1 and p400 is applicable to chromatin remodeling at promoters controlled by TIP60. Aim 3 will test the role of ATP binding/hydrolysis and oligomerization with self or with RVB2. In vitro assays will be developed to explore the molecular basis of how RVB1 activates the TIP60 acetyltransferase complex and how the TIP60 complex mobilizes phosphoH2AX-containing nucleosomes prior to dephosphorylation of phosphoH2AX. PUBLIC HEALTH RELEVANCE The cellular response to DNA damage determines both how environmental DNA damaging agents lead to mutations and cause cancers and how therapeutic DNA damaging agents treat cancers. Chromatin remodelers regulated by RVB act both at sites of DNA damage and at promoters to control the cell's response. Therefore a molecular understanding of how RVB regulates chromatin remodeling factors important for the optimal response to DNA damage will allow the design of chemicals that interfere with RVB function and thus sensitize cells to chemo- or radio-therapy.

描述（由申请人提供）：人类细胞对 DNA 损伤反应的两个重要分子特征是组蛋白变体 H2AX 的磷酸化和转录程序的变化。我们实验室的最新结果表明 RVB1 蛋白与细胞对 DNA 损伤的反应有关。人类 RVB 是染色质重塑复合物的组成部分，该复合物含有组蛋白乙酰转移酶、TIP60、ATPase p400 和其他几种细胞蛋白。 RVB1 或 TIP60 耗竭会导致 DNA 损伤后磷酸化 H2AX 积累增加，并使癌细胞对 DNA 损伤剂敏感，这表明 RVB ATP 酶抑制剂可能在化疗或放疗中有用。该项目将测试两个假设：1) RVB1 需要中和 p400 对 TIP60 复合物的乙酰转移酶活性的抑制作用，2) TIP60 的乙酰转移酶活性需要在 DNA 去磷酸化之前将含有核小体的 phosphoH2AX 从 DNA 上移走。细胞磷酸酶。目标 1 将探讨 TIP60 是否是参与下调磷酸化 H2AX 的主要染色质重塑因子，以及 RVB2 是否与 RVB1 在此功能中合作。目标 2 将测试 RVB1 和 p400 调节 TIP60 的假设是否适用于 TIP60 控制的启动子处的染色质重塑。目标 3 将测试 ATP 结合/水解以及自身或 RVB2 寡聚化的作用。将开发体外测定来探索RVB1如何激活TIP60乙酰转移酶复合物以及TIP60复合物如何在磷酸化H2AX去磷酸化之前动员含有磷酸化H2AX的核小体的分子基础。公众健康相关性细胞对 DNA 损伤的反应决定了环境 DNA 损伤剂如何导致突变并引发癌症，以及治疗性 DNA 损伤剂如何治疗癌症。 RVB 调节的染色质重塑因子在 DNA 损伤位点和启动子处发挥作用，以控制细胞的反应。因此，从分子角度理解 RVB 如何调节染色质重塑因子（对于 DNA 损伤的最佳反应至关重要），将有助于设计干扰 RVB 功能的化学物质，从而使细胞对化疗或放疗敏感。