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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：
8164682
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 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8164682
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. PUBLIC HEALTH RELEVANCE: During the formation of sperm and eggs, chromosomes must segregate accurately. However, approximately 10% of clinically reported pregnancies are chromosomally abnormal, resulting in pregnancy loss or genetic defects such as Down syndrome. Very little is known about the genetic failures that cause these chromosomal abnormalities in sex cells; however, we will significantly enhance our knowledge of this phenomenon by achieving the goals set within this proposal.

描述（由申请人提供）：我们知道减数分裂期间受到干扰的DNA损伤反应将导致不孕，妊娠丢失或遗传缺陷。然而，我们对减数分裂过程中DNA损伤反应的调控知之甚少。我的初步数据表明SMC5/6复合体在减数分裂细胞周期进程中介导DNA损伤反应是必需的。由于SMC5/6复合体对细胞活力至关重要，因此尚未在哺乳动物体内进行实验，我们对SMC5/6在哺乳动物中的功能的了解有限。我正在制造Smc5的生殖细胞特异性突变并使睾丸特异性Smc5 /6基因Eid3发生突变。这些突变体将使我能够在减数分裂（Aim1）中进行SMC5/6复合体功能的首次全面研究。EID3是SMC5/6复合体的睾丸特异性卵磷脂亚基。然而，体细胞蛋白酶SMC5/6亚基NSE4也在睾丸内表达。因此，在睾丸内存在两种SMC5/6复合物，SMC5/6NSE4和SMC5/6EID3。我将确定这两个综合体之间的异同（目标2）。我已经确认SMC5/6复合物与BAT3-EP300复合物相互作用。BAT3-EP300复合体是激活trp53介导的DNA损伤反应所必需的。根据SMC5/6与BAT3-EP300复合物之间形成的蛋白质相互作用网络，SMC5/6通过调节TRP53乙酰化而成为DNA损伤应答通路的关键组成部分。在Aim 3中，我将详细评估SMC5/6与BAT3-EP300之间的相互作用，并对EID3对BAT3-EP300介导的TRP53乙酰化的拮抗作用进行生化分析。我通过假设SMC5/6作为修复/监视复合体协调DNA修复和trp53介导的DNA损伤反应来接近K99/R00奖励。这个功能保证了减数分裂前期到中期转换的保真度。