The SMC5/6 Complex - DNA Damage Response Regulation Ensures Meiotic Fidelity

SMC5/6 复合物 - DNA 损伤反应调节确保减数分裂保真度

基本信息

批准号：
8321387
负责人：
Philip W Jordan
金额：
$ 9万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-19 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8321387
关键词：
Acetylation Address Alleles Animal Model Antibodies Award Biochemical Biological Assay Cell Cycle Progression Cell Survival Chromatin Chromosome Segregation Chromosome abnormality Chromosomes Chromosomes, Human, Pair 5 Complex DNA Damage DNA Repair Data Development Differentiation Inhibitor Down Syndrome EP300 gene Ensure Event Exons Failure Female Fertilization Gametogenesis Genes Genetic Genetic Variation Germ Cells Goals Immunoprecipitation Infertility Investigation Knock-out Knockout Mice Knowledge Maintenance Mammals Mediating Meiosis Meiotic Prophase I Meiotic Recombination Mentors Metaphase Mus Mutate Mutation Pathway interactions Phase Play Pregnancy Pregnancy loss Process Property Prophase Proteins Regulation Reporting Role Somatic Cell Spermatogenesis Synaptonemal Complex TP53 gene Techniques Testing Testis Work Yeasts design egg in vivo inhibitor/antagonist male meetings mutant novel recombinase repaired research study response sexual dimorphism sperm cell yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): We know a perturbed DNA damage response during meiosis will result in infertility, pregnancy loss or genetic defects. However, we know very little about the regulation of the DNA damage response during meiosis. My preliminary data implies that the SMC5/6 complex is required to mediate the DNA damage response during meiotic cell cycle progression. Because the SMC5/6 complex is essential for cell viability, in vivo experiments in mammals have not been performed and our understanding of the function of SMC5/6 in mammals is limited. I am creating a germ cell-specific mutation of Smc5 and mutating the testis specific SMC5/6 gene Eid3. These mutants will enable me to perform the first comprehensive studies of SMC5/6 complex function in meiosis (Aim1). EID3 is a testis-specific kleisin subunit of the SMC5/6 complex. However, the somatic cell kleisin SMC5/6 subunit, NSE4, is also expressed within the testis. Therefore, there are two SMC5/6 complexes present within the testis, SMC5/6NSE4 and SMC5/6EID3. I will determine the similarities and differences between the two complexes (Aim 2). I have confirmed that the SMC5/6 complex interacts with the BAT3-EP300 complex. The BAT3-EP300 complex is required for the activation of a TRP53-mediated DNA damage response. According to the protein interaction network developed between the SMC5/6 and the BAT3-EP300 complexes, SMC5/6 emerges as a key component of the DNA damage response pathway, by regulating TRP53 acetylation. In Aim 3, I will conduct a detailed assessment of the interaction between SMC5/6 and BAT3-EP300, together with biochemical analysis of the antagonistic function of EID3 on BAT3-EP300-mediated TRP53 acetylation. I approach the K99/R00 award with the hypothesis that SMC5/6 acts as a repair/surveillance complex coordinating DNA repair and the TRP53-mediated DNA damage response. This function ensures fidelity at the prophase to metaphase I transition of meiosis.

描述（由申请人提供）：我们知道减数分裂过程中受干扰的DNA损伤反应将导致不育症，妊娠丧失或遗传缺陷。但是，我们对减数分裂过程中DNA损伤反应的调节知之甚少。我的初步数据意味着需要SMC5/6复合物在减数分裂细胞周期进程中介导DNA损伤反应。由于SMC5/6复合物对于细胞活力是必不可少的，因此尚未进行哺乳动物的体内实验，并且我们对SMC5/6在哺乳动物中的功能的理解受到限制。我正在创建SMC5的生殖细胞特异性突变，并突变睾丸特异性SMC5/6基因EID3。这些突变体将使我能够对减数分裂中的SMC5/6复杂功能进行首次全面研究（AIM1）。 EID3是SMC5/6复合物的睾丸特异性kleisin亚基。但是，在睾丸中也表达了体细胞kleisin smc5/6亚基NSE4。因此，睾丸中存在两个SMC5/6复合物，SMC5/6NSE4和SMC5/6EID3。我将确定两个复合物之间的相似性和差异（AIM 2）。我已经确认SMC5/6复合物与BAT3-EP300复合物相互作用。激活TRP53介导的DNA损伤响应需要BAT3-EP300复合物。根据SMC5/6和BAT3-EP300复合物之间开发的蛋白质相互作用网络，SMC5/6通过调节TRP53乙酰化而成为DNA损伤响应途径的关键组成部分。在AIM 3中，我将对SMC5/6和BAT3-EP300之间的相互作用进行详细评估，并对EID3对BAT3-EP300介导的TRP53乙酰化的拮抗功能的生化分析。我通过假设SMC5/6作为修复/监视复合物协调DNA修复和TRP53介导的DNA损伤响应的假设。此功能可确保对中期I过渡的预言性的保真度。