Regulation and Localization of Mismatch Repair Proteins

错配修复蛋白的调控和定位

• 批准号: 10704811
• 负责人: Joanna E Haye
• 金额: $ 5.3万
• 依托单位: XAVIER UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 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/10704811
• 关键词: Acetylation; Affect; Binding; Biological Assay; Chromatin; Colon; Complex; DNA; DNA biosynthesis; Defect; Dependence; Dimerization; Endometrial Carcinoma; Equipment; 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; Methylation; Methyltransferase; Microsatellite Instability; Mismatch Repair; 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; Role; Saccharomyces cerevisiae; Site; Small Intestines; Syndrome; Testing; Tissues; Two-Hybrid System Techniques; Urinary tract; Variant; Work; Yeasts; cancer type; chromatin immunoprecipitation; demethylation; early onset colon cancer; experimental study; gene repair; genome integrity; histone methylation; histone modification; overexpression; parent grant; programs; recruit; repair function

Abstract: An equipment supplement is requested to perform the experiments summarized below. This is necessary to perform the experiments outlined in the parent grant. DNA mismatch repair (MMR) is a highly conserved process. A functional MMR pathway is essential for maintaining genome integrity and 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, and upper urinary tract as well as other tissues. In our most recent publication, we showed that deletion of Modulator of Transcription (Not4) and General Control Nonderepressible 5 (Gcn5) modulate the levels of MutSα (consisting of Msh2 and Msh6), the major complex involved in MMR. Not4 and Gcn5 are proteins that ubiquitylate and acetylate various proteins respectively. We hypothesize that these proteins modify MutSα and that the modifications affect the stability and localization of the complex. Further studies need to be conducted to gain a better understanding of how Not4 and Gcn5 regulate these MMR proteins. Additionally, our previous experiments have shown that yeast MutSα is in the vicinity of the replication machinery during DNA replication. Human MutSα interacts with the replication machinery by binding PCNA and also recognizes specific histone modifications. How yeast MutSα is recruited to chromatin remains elusive. Using the yeast Saccharomyces cerevisiae (S. cerevisiae), this project aims to: 1) further examine the role of Gcn5 and Not4 in the regulation of MutSα stability and 2) determine the effect of post-translational modifications on MMR protein recruitment to chromatin.

抽象的： 要求设备补充以执行以下概述的实验。这是 进行父母赠款中概述的实验所需的必要条件。 DNA不匹配维修（MMR）是 高度保守的过程。功能性MMR途径对于维持基因组完整性至关重要 MMR的丧失会导致较高的真核生物中的基因组不稳定性和癌症。例如，缺陷 在MMR基因中导致林奇综合征，一种常见的遗传癌综合征，导致早期 结肠，子宫内膜，卵巢，小肠，肝胆道和上部的发病癌症 尿路以及其他组织。在我们最近的出版物中，我们表明了删除 转录调节剂（NOT4）和通用对照不可用的5（GCN5）调节 MUTSα的水平（由MSH2和MSH6组成），这是MMR涉及的主要复合物。 NOT4和GCN5 是分别泛素酯和乙酰化各种蛋白质的蛋白质。我们假设这些 蛋白质修饰MUTSα，并且修饰会影响复合物的稳定性和定位。 需要进行进一步的研究，以更好地了解NOT4和GCN5如何调节 这些MMR蛋白。此外，我们以前的实验表明酵母菌α在 DNA复制过程中复制机制附近。人类Mutsα与 通过结合PCNA并识别特定的组蛋白修饰，复制机制。多么酵母 将MUTSα募集到染色质中保持弹性。使用酿酒酵母的酵母菌（S. 酿酒酵母），该项目的目的是：1）进一步研究GCN5和NOT 4在调节中的作用 MUTSα稳定性和2）确定翻译后修饰对MMR蛋白的影响 招募染色质。