Roles of Checkpoint and DNA Repair Proteins in Fission Yeast Telomere Maintenance

检查点和 DNA 修复蛋白在裂殖酵母端粒维护中的作用

基本信息

批准号：
7259441
负责人：
Toru Nakamura
金额：
$ 26.33万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7259441
关键词：
Affect Apoptosis Biological Assay Biological Models Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cells Chromatin Structure Chromosomes Complex DNA DNA Damage DNA Repair DNA Repair Enzymes DNA Structure DNA biosynthesis DNA damage checkpoint DNA repair protein Development Event Fission Yeast Future Generations Genetic Recombination Genome Genomic Instability Goals Hereditary Disease Histone H2A Homologous Gene Human Length Link Methods Modeling Mutation Phosphorylation Phosphotransferases Play Primer Extension Protein Binding Proteins Recruitment Activity Reporter Research Research Personnel Role Schizosaccharomyces pombe Proteins Shapes Southern Blotting Structure Telomerase Telomere Maintenance Telomere-Binding Proteins Testing Transcriptional Activation Up-Regulation Work base human study insight mutant neoplastic cell prevent programs research study response sensor telomere tumor tumor growth tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Proper maintenance of telomere structure is crucial for stable inheritance of the genome. Various checkpoint and DNA repair proteins, including evolutionarily highly conserved checkpoint kinases Tel1 (ATM) and Rad3 (ATR), play important roles in stable maintenance of telomeres. However, no clear mechanistic roles for various checkpoint and DNA repair proteins in telomere maintenance have been established. Major goal of our research is to understand how checkpoint and DNA repair proteins contribute to telomere maintenance. The current proposal will utilize fission yeast Schizosaccharomyces pombe as a model system. Highly conserved DNA damage responses and telomere maintenance mechanisms between fission yeast and humans should be helpful in extrapolating our findings to build testable models for human cells. Deregulation of telomere maintenance mechanisms has been found to be a key event in tumorigenesis, thus mechanistic insights on how various proteins collaborate to generate functional telomeres are first needed to devise effective methods for preventing tumorigenesis. In order to be stably maintained, telomeres must fulfill two major functions. First, telomeres must protect telomeric DNA ends from fusions and degradation. Second, telomeres must provide access to telomerase to prevent loss of telomeric DNA after DNA replication. Thus, telomeres must undergo dynamic switches from the highly protected state to the more accessible state that allows recruitment of telomerase. We hypothesize that checkpoint and DNA repair proteins are recruited to telomeres in a cell cycle-regulated manner to trigger appropriate changes in telomere structure and telomere protein composition. This model will be directly tested in Aim 1. Additionally, newly developed fission yeast reporter strains will be utilized to search for potential telomere targets of Rad3 and Tel1 kinases in Aim 2. Our preliminary studies with MRN (Mre11-Rad50-Nbs1) complex mutants have suggested that recruitment of Tel1 to telomeres does not require Tel1-MRN interaction unlike recruitment of Tel1 to other DNA breaks. Therefore, attempts will be made in Aim 3 to understand the mechanistic basis for the MRN-independent recruitment of Tel1 to telomeres.

描述（由申请人提供）：端粒结构的适当维护对于基因组的稳定遗传至关重要。各种检查点和DNA修复蛋白，包括进化上高度保守的检查点激酶Tel1（ATM）和RAD3（ATR），在稳定维持端粒中起重要作用。但是，尚未确定各种检查点和DNA修复蛋白在端粒维持中的明确机械作用。我们研究的主要目标是了解检查点和DNA修复蛋白如何有助于端粒维护。当前的提案将利用裂变酵母菌菌果糖pombe作为模型系统。高度保守的DNA损伤反应和裂变酵母和人之间的端粒维持机制应有助于推断我们的发现以构建人类细胞可测试的模型。发现端粒维持机制的放松管制是肿瘤发生中的关键事件，因此首先需要关于各种蛋白质如何共同生成功能端粒的机械见解，以设计有效的方法来预防肿瘤发生。为了稳定维护，端粒必须履行两个主要功能。首先，端粒必须保护端粒DNA末端免受融合和降解。其次，端粒必须提供端粒酶的访问，以防止DNA复制后端粒DNA的损失。因此，端粒必须从高度保护状态到更容易访问的状态，以募集端粒酶。我们假设检查点和DNA修复蛋白以细胞周期调节的方式募集到端粒，以触发端粒结构和端粒蛋白组成的适当变化。 This model will be directly tested in Aim 1. Additionally, newly developed fission yeast reporter strains will be utilized to search for potential telomere targets of Rad3 and Tel1 kinases in Aim 2. Our preliminary studies with MRN (Mre11-Rad50-Nbs1) complex mutants have suggested that recruitment of Tel1 to telomeres does not require Tel1-MRN interaction unlike recruitment of Tel1 to other DNA breaks.因此，将尝试以目标3进行尝试，以了解将Tel1独立招募到端粒的机械基础。