Roles for Mismatch Repair Proteins in Maintaining Genome Stability

错配修复蛋白在维持基因组稳定性中的作用

• 批准号: 10591126
• 负责人: Eric E. Alani
• 金额: $ 1.02万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10591126
• 关键词: Alleles; Chromosomal Rearrangement; Chromosome Segregation; Complex; Confocal Microscopy; DNA; DNA Sequence; DNA biosynthesis; Disease; Doctor of Philosophy; Ensure; Eukaryota; Fluorescence; Fluorescence Microscopy; Future; Genetic Crossing Over; Genetic Recombination; Genome Stability; Group Meetings; Hereditary Nonpolyposis Colorectal Neoplasms; Infertility; Journals; Kinetics; Lead; Learning; Link; Location; Malignant Neoplasms; Measures; Meiosis; Mentors; Mismatch Repair; Molecular; Molecular Biology; Mutation; Nuclear Pore; Oral; Participant; Pathogenicity; Protein Family; Proteins; RAD52 gene; Research; Role; Saccharomyces cerevisiae; Site; Statistical Data Interpretation; System; Time; Western Blotting; Work; base; colon cancer patients; experience; experimental study; gene repair; helicase; homologous recombination; insertion/deletion mutation; interest; parent grant; prevent; programs; recruit; repaired; summer research; tool; undergraduate research experience

Project Summary DNA mismatch repair (MMR) systems act to excise misincorporation errors that occur during DNA replication. In eukaryotes MSH proteins recognize these errors in the context of base-base and insertion/deletion mismatches and recruit MLH complexes to form ternary complexes that work with replication factors and Exo1 to excise the newly replicated DNA strand through the mismatch site. This is followed by DNA re-synthesis steps. MMR factors also recognize mismatches that form during strand invasion steps in homologous recombination; they recruit a helicase complex that unwinds (rejects) recombination intermediates and allows a new homology search. Sydney Rosen's summer research is aimed at understanding the decision steps that take place during homologous recombination when a double-strand break occurring in a DNA sequence is repaired using a donor sequence containing low levels of sequence divergence (Project 3 in parent grant R35GM134872). More specifically, she is interested in understanding how the kinetics and subnuclear location(s) of homologous recombination in S. cerevisiae compare when an identical donor sequence is used as a template for repair versus a divergent donor template. She plans to track and measure the timing of repair of an induced DSB via a fluorescence-marked protein involved in homologous recombination (Rad52) as well as nuclear pore components. Her hypothesis is that recombination between divergent sequences will result in slower repair and possibly differences in repair location. Results from this time course will provide a better understanding of how the rejection decision is facilitated, and how chromosomal rearrangements can lead to disease. Sydney has expressed strong interest in entering a Ph.D. program in molecular biology after graduating from Cornell. She will become experienced transforming baker's yeast and analyzing transformants using western blot analysis and fluorescence and confocal microscopy. She will also learn the appropriate statistical analyses for each of her experiments. Her mentors will meet with her daily to discuss her research progress and plan future experiments. She is already an active participant in group meeting and journal club, presenting research summaries and formal presentations. Over the summer Sydney will join our department's summer Research Experience for Undergraduates (REU) program where she will gain experience presenting her work in written and oral formats.

项目概要 DNA 错配修复 (MMR) 系统可消除 DNA 复制过程中发生的错误掺入错误。 在真核生物中，MSH 蛋白在碱基和插入/缺失的背景下识别这些错误 错配并招募 MLH 复合物形成与复制因子和 Exo1 一起作用的三元复合物 通过错配位点切除新复制的 DNA 链。随后进行 DNA 重新合成 步骤。 MMR 因子还可以识别同源链入侵步骤中形成的错配。 重组；他们招募解旋酶复合物，解旋（拒绝）重组中间体并允许 新的同源性搜索。西德尼·罗森 (Sydney Rosen) 的夏季研究旨在了解以下决策步骤： 当 DNA 序列中发生双链断裂时，同源重组过程中就会发生 使用包含低水平序列分歧的供体序列进行修复（父资助中的项目 3） R35GM134872）。更具体地说，她有兴趣了解动力学和亚核如何 当使用相同的供体序列时，比较酿酒酵母中同源重组的位置 作为修复模板与发散供体模板的比较。她计划跟踪和衡量修复的时间 通过参与同源重组的荧光标记蛋白 (Rad52) 诱导 DSB 作为核孔成分。她的假设是不同序列之间的重组将导致 修复速度较慢，并且修复位置可能存在差异。本次课程的结果将提供更好的 了解如何促进排斥决定，以及染色体重排如何导致 疾病。悉尼表达了攻读博士学位的浓厚兴趣。分子生物学课程后 毕业于康奈尔大学。她将在转化面包酵母和分析转化体方面变得经验丰富 使用蛋白质印迹分析以及荧光和共聚焦显微镜。她还将学习适当的 对她的每个实验进行统计分析。她的导师每天都会与她会面讨论她的研究 进展并计划未来的实验。她已经是小组会议和期刊俱乐部的积极参与者， 提出研究摘要和正式演示。整个夏天，悉尼将加入我们部门的 本科生暑期研究经验 (REU) 项目，她将获得演讲经验 她的作品有书面和口头形式。