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: 10619197
• 负责人: Eric E. Alani
• 金额: $ 2.8万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10619197
• 关键词: Alleles; Area; Biochemical; Chromosomal Rearrangement; Chromosome Segregation; Collection; Colon Carcinoma; Communication; Complex; DNA; DNA Sequence; DNA biosynthesis; Disease; Early Diagnosis; Educational process of instructing; Educational workshop; Effectiveness; Ensure; Eukaryota; Funding; Genetic Crossing Over; Genetic Recombination; Genome; Genome Stability; Goals; Group Meetings; Hereditary Nonpolyposis Colorectal Neoplasms; Individual; Infertility; Inosine Diphosphate; Interest Group; Laboratories; Lead; Link; Literature; MLH1 gene; Malignant Neoplasms; Maps; Meiosis; Mentors; Mismatch Repair; Molecular; Molecular Evolution; Monitor; Mutation; Mutation Analysis; PMS1 gene; Pathogenicity; Patients; Protein Family; Proteins; Reading; Research; Research Training; Role; Saccharomyces cerevisiae; Science; Scientist; Site; System; TimeLine; Training; Universities; Work; base; career; diagnostic tool; effectiveness evaluation; gene repair; graduate student; insertion/deletion mutation; interest; meetings; novel diagnostics; prevent; recruit; repair function; symposium; tool

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. Mutations in MMR genes have been found in about half of individuals suffering from hereditary non-polyposis colorectal cancer, underscoring the importance of obtaining new mechanistic understandings and molecular tools to establish allele pathogenicity. This diversity supplement to R35GM134872 is aimed at developing and executing a mentoring plan for a first- year graduate student. This plan focuses on two components that will be implemented in the funding period that involve research training and professional mentoring. The research plan was developed by the trainee after reading the MMR literature and initiating discussions with the mentor, co-mentor, and scientists at Cornell interested in genome stability and molecular evolution. The trainee then decided to focus on the question of how subunits of a critical complex in MMR, Mlh1-Pms1, have co-evolved to perform MMR functions in contrast to other MLH complexes, which act primarily in meiotic genetic recombination. The trainee proposes to identify reversion mutations in the baker’s yeast PMS1 gene that partially or wholly restore compatibility with one of four mlh1 mutations previously identified by the Alani laboratory that confer MMR function in one strain background but not in a divergent one. Results from this work in conjunction biochemical analyses are expected to provide an understanding of related functional areas within heterodimeric complexes. More importantly, through an analysis of the map generated by mutational analysis in conjunction with a study of co-evolving regions of MMR genes from a world-wide collection of 1,010 baker’s yeast genomes, the trainee expects to develop an understanding of how the MLH1 and PMS1 genes have diverged from other MLH complexes to specifically act in MMR. The mentoring plan was developed after the trainee and advisor completed an IDP which recognizes the trainee’s career goals and the training required to meet them. An individualized plan was then developed to ensure that the trainee attends professional workshops provided by Cornell University focused on teaching and science communication, as well as research conferences. A timeline was created to monitor progress on the plan and to assess the effectiveness of mentoring at different stages in the funding period. The effectiveness of these opportunities will be assessed through weekly meetings with the advisor, committee meetings, and group meetings that involve the Alani laboratory and other groups interested in genome stability on campus.

DNA错配修复（MMR）系统用于切除DNA错配过程中发生的错误， 复制的在真核生物中，MSH蛋白在碱基-碱基和碱基-碱基的背景下识别这些错误。 插入/缺失错配和募集MLH复合物以形成三元复合物， 复制因子和Exo 1通过错配位点切除新复制的DNA链。这是 然后进行DNA再合成步骤。MMR基因的突变已经在大约一半的人中发现。 患有遗传性非息肉病性结直肠癌的人，强调了 获得新的机制理解和分子工具来建立等位基因致病性。这 R35 GM 134872的多样性补充旨在制定和执行第一个- 年毕业生。该计划的重点是两个组成部分，将实施的资金 期间，涉及研究培训和专业辅导。研究计划由 学员在阅读MMR文献并与导师、共同导师进行讨论后， 康奈尔大学的科学家对基因组稳定性和分子进化感兴趣。学员于是决定， 关注MMR中关键复合物Mlh 1-Pms 1的亚基如何共同进化， 与其他主要在减数分裂遗传学中起作用的MLH复合物相比，MMR执行MMR功能。 重组受训者提出鉴定面包酵母PMS 1基因中的回复突变， 部分或全部恢复与Alani先前鉴定的四种mlh 1突变之一的相容性， 在一个菌株背景中赋予MMR功能但在不同菌株背景中不赋予MMR功能实验室。结果 这项工作结合生物化学分析，预计将提供相关的理解， 异二聚体复合物内的功能区。更重要的是，通过分析地图 通过突变分析以及对来自一个人的MMR基因的共同进化区域的研究产生的， 在收集了世界范围内的1,010个面包酵母基因组后， MLH 1和PMS 1基因如何从其他MLH复合物中分化出来，从而特异性地作用于MMR。 指导计划是在受训者和顾问完成一份IDP后制定的，该IDP承认 学员的职业目标以及实现这些目标所需的培训。一个个性化的计划， 旨在确保受训者参加康奈尔大学提供的专业研讨会 专注于教学和科学交流，以及研究会议。一条时间轴 设立该委员会的目的是监测计划的进展情况，并评估不同阶段指导的有效性 在融资期间。将通过每周会议评估这些机会的有效性 与顾问，委员会会议和小组会议，涉及阿拉尼实验室和其他 对基因组稳定性感兴趣的团体