Regulation and Localization of Mismatch Repair Proteins

错配修复蛋白的调控和定位

基本信息

批准号：
10671757
负责人：
Joanna E Haye
金额：
$ 18.75万
依托单位：
XAVIER UNIVERSITY OF LOUISIANA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10671757
关键词：
Acetylation Affect Binding Biological Assay Chromatin Colon Complex DNA DNA biosynthesis DNA-Directed DNA Polymerase Defect Dependence Dimerization Endometrial Carcinoma Eukaryota Frequencies Genetic Transcription Genomic Instability Hepatobiliary Hereditary Neoplastic Syndromes Hereditary Nonpolyposis Colorectal Neoplasms Histones Human Immunoblot Analysis Investigation Lysine Maintenance Malignant Neoplasms Malignant neoplasm of ovary Measures Methylation Methyltransferase Microsatellite Instability Mismatch Repair Mismatch Repair Deficiency Modeling Modification Mutagenesis Mutagens Mutation Pathway interactions Polymerase Chain Reaction Post-Translational Protein Processing Principal Investigator Process Prokaryotic Cells Proteins Proteomics Publications Publishing Regulation Repair Complex Research Role Saccharomyces cerevisiae Site Small Intestines Syndrome Testing Tissues Two-Hybrid System Techniques Urinary tract Work Yeasts cancer type chromatin immunoprecipitation demethylation early onset colon cancer experimental study gene repair genetic manipulation genetic predictors genome integrity histone methylation histone modification mutant overexpression programs recruit repair function

项目摘要

Principal Investigator/Program Director (Last, first, middle): Haye, Joanna, Elizabeth Abstract: DNA mismatch repair (MMR) is a highly conserved process. A functional MMR pathway is essential for maintaining genome integrity; loss of MMR results in genome instability and cancer in higher eukaryotes. For example, defects in MMR genes result in Lynch Syndrome, a common hereditary cancer syndrome resulting in early onset cancers of the colon, endometrium, ovaries, small intestine, hepatobiliary tract, upper urinary tract as well as other tissues. In our most recent publication, we showed that in yeast, deletion of Modulator of Transcription (also known as Not4) or General Control Nonderepressible 5 (Gcn5) modulate the levels of Msh2, a major MMR component. Loss of Gcn5 significantly decreases Msh2, whereas deleting Not4 stabilizes functional Msh2. Not4 and Gcn5 are proteins that ubiquitylate and acetylate various proteins respectively. We hypothesize that Not4 and Gcn5 modify yeast MutSα (comprised of Ms2 and Msh6) and that the modifications affect the stability of the complex. Using the yeast Saccharomyces cerevisiae (S. cerevisiae), the first aim of the proposed research is to establish the role of Gcn5 and Not4 in the regulation of the major mismatch recognition complex MutSα. Our previous experiments have also shown that yeast MutS tracks with the replication machinery during DNA replication. Human MutSα is recruited to chromatin through specific histone modifications and interacts with the replication machinery by binding PCNA, the DNA polymerase processivity factor. However, the modifications that recruit human MutSα are not utilized in yeast. How yeast MutSα is recruited to chromatin remains elusive. The second aim of this research is to determine the role of post- translational modifications in MutSα recruitment to chromatin. PHS398 (Rev. 5/01) Page Continuation Format Page

首席研究员/计划主任（最后，第一，中间）：海耶，乔安娜，伊丽莎白抽象的： DNA不匹配修复（MMR）是一个高度保守的过程。功能性MMR途径对于维持基因组完整性； MMR的丧失导致较高的真核生物中的基因组不稳定性和癌症。为了例如，MMR基因缺陷导致林奇综合征，这是一种常见的遗传癌综合征，导致结肠，子宫内膜，卵巢，小肠，肝胆道，上尿路的早期发病癌症以及其他组织。在我们最近的出版物中，我们表明，在酵母中，删除了调节器的删除转录（也称为非4）或通用对照不可抑制5（GCN5）调节水平 MSH2，主要的MMR组件。 GCN5的丢失显着降低了MSH2，而删除NOT4稳定功能MSH2。 NOT4和GCN5是分别泛素酯和乙酰化各种蛋白质的蛋白质。我们假设NOT4和GCN5修改了酵母Mutsα（由MS2和MSH6组成），并且修改影响复合物的稳定性。使用酿酒酵母（酿酒酵母）的酵母菌，第一个目的拟议的研究是确定GCN5和NOT 4在主要不匹配的调节中的作用识别复合物mutsα。我们以前的实验还表明，酵母菌与 DNA复制过程中的复制机制。通过特异性组蛋白募集人Mutsα 通过结合PCNA（DNA聚合酶加工性），修饰并与复制机械相互作用因素。但是，募集人Mutsα的修饰并未在酵母中使用。酵母Mutsα如何招募到染色质仍然难以捉摸。这项研究的第二个目的是确定后的作用 MUTSα募集到染色质中的翻译修饰。 PHS398（Rev. 5/01）页面延续格式页面