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：
8118466
负责人：
Anindya Dutta
金额：
$ 28.95万
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 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8118466
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 Cisplatin Complex DNA DNA Damage DNA Double Strand Break Down-Regulation Etoposide Fingers Gene Expression Gene Silencing Genes HTATIP gene Health Histone Acetylation Histone H4 Histones Homologous Gene Human Hydrolysis Laboratories Lead Malignant Neoplasms Measures Metabolism Mitomycins Molecular Molecular Profiling Mutate Mutation Nucleosomes Phenotype Phosphoric Monoester Hydrolases Phosphorylation Play Protein Dephosphorylation Proteins Publishing Radiation therapy Radio Recruitment Activity Regulation Relative (related person)Role Site Structure Testing Therapeutic Transferase UV induced DNA damage Up-Regulation Variant Walkers Yeasts base cancer cell cancer therapy chemotherapy chromatin remodeling design gene discovery histone acetyltransferase human H2AX 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暗示了RVB1蛋白在细胞对DNA损伤的反应中。人RVB是包含组蛋白乙酰转移酶TIP60，ATPase P400和其他几种细胞蛋白的染色质重塑复合物的组成部分。 RVB1或TIP60耗竭会导致DNA损伤后磷酸2AX的积累增加，并使癌细胞对DNA损伤剂敏感，这表明RVB ATPase的抑制剂可能对化学或放射疗法有用。该项目将检验两个假设：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和2）2）TIP60的乙酰基转移酶活性需要在含DNA的核糖体之前通过其Dephosphohate phosphospass of phosphoH2AX动员phosphoH2AX，该核体在其dephosphosph of cellosphosphalation phospassase之前含有DNA。 AIM 1将探索TIP60是否是调节PhosphoH2AX的主要染色质重塑，以及RVB2是否在此功能中与RVB1合作。 AIM 2将测试RVB1和P400对TIP60调控的假设是否适用于由TIP60控制的启动子对染色质的重塑。 AIM 3将测试与自我或RVB2的ATP结合/水解和低聚的作用。将开发体外测定，以探索RVB1如何激活TIP60乙酰转移酶复合酶复合物的分子基础，以及在磷酸化磷酸化之前，TIP60如何动员含PhosphoH2AX的核小体。公共卫生相关性的细胞对DNA损伤的反应决定了环境DNA损伤剂如何导致突变和导致癌症以及治疗性DNA损害剂如何治疗癌症。由RVB ACT调节的染色质重塑剂在DNA损伤部位和启动子的位置都可以控制细胞的反应。因此，对RVB如何调节染色质重塑因子的分子理解对于对DNA损伤的最佳反应很重要，将允许设计干扰RVB功能的化学物质，从而使细胞对化学或放射治疗敏感。