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.

摘要： 需要补充设备来执行下面总结的实验。这是 执行父母拨款中概述的实验所必需的。DNA错配修复(MMR)是一种 高度保守的过程。一个有功能的MMR途径对于维持基因组的完整性是必不可少的 而MMR的缺失会导致基因组的不稳定和高等真核生物的癌症。例如，缺陷 MMR基因导致Lynch综合征，这是一种常见的遗传性癌症综合征，导致早期 起病的结肠癌、子宫内膜癌、卵巢癌、小肠癌、肝胆管癌和上段癌 尿路和其他组织。在我们最新出版物中，我们显示了删除 转录调节因子(Not4)和一般控制不可降压因子5(Gcn5)调节 参与MMR的主要复合体MutSα(由Msh2和Msh6组成)的水平。Not4和Gcn5 泛素化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，这些修饰影响复合体的稳定性和定位。 需要进行进一步的研究，以更好地了解Not4和Gcn5是如何调节的 这些MMR蛋白。此外，我们之前的实验表明，酵母MutSα在 在DNA复制过程中复制机器的附近。人类MutSα与 通过结合增殖细胞核抗原的复制机制，也识别特定的组蛋白修饰。如何酿造酵母 MutSα被招募到染色质仍然难以捉摸。利用酿酒酵母(S. ，这个项目的目的是：1)进一步研究Gcn5和Not4在调节 MutSα稳定性和2)确定翻译后修饰对MMR蛋白的影响 重新招募到染色质。