Role of GCNA in preserving genome integrity and fertility

项目摘要 我们提出了一种跨物种的方法来了解维持基因组完整性的途径， 细胞物种的延续需要基因组受到保护，不受内源性和外源性的影响。 威胁这一点在生殖细胞（卵子和精子的前体）中最为关键。缺陷 警惕途径导致基因组完整性丧失和后代活力的随后恶化，和/或 流产一种新发现的高度保守的遗传因子GCNA（生殖细胞核因子）的详细研究 酸性肽酶），将提供新的见解基因组完整性和生育力的关键决定因素。我们 初步数据显示GCNA的缺失导致果蝇和C.优雅， 表明功能的保守性。这里概述的实验将提供一些初步的见解 保守的GCNA家族成员通过共同的机制促进基因组的完整性和生育力。 物种我们发现GCNA既与拓扑异构酶2相互作用，又阻止DNA：蛋白质的形成。 交联（DPC）。GCNA突变动物表现出增加的染色体丢失、突变积累， 细胞周期缺陷，将GCNA置于多个基因组完整性过程的中心。探讨 gcna在生殖细胞维持中的功能机制，我们提出了三个具体目标：1） 分析染色体丢失的病因，并确定gcna对细胞周期调控的贡献，2） 表征与拓扑异构酶2的相互作用，鉴定保守的结合伴侣，并阐明 DPC的性质和病因学，以及3）了解GCNA的结构域及其功能相关性 对人类生育能力的影响

PROJECT SUMMARY We propose a cross-species approach to understand the pathways that maintain genome integrity in germ cells. The perpetuation of the species requires that the genome is protected from endogenous and exogenous threats. Nowhere is this more critical than in the germ cells, the precursors of egg and sperm. Defects in the vigilance pathways lead to loss of genome integrity and subsequent deterioration of offspring vigor and/or miscarriage. Detailed studies of a newly identified, highly conserved genetic factor, GCNA (Germ Cell Nuclear Acidic Peptidase), will provide novel insights into key determinants of genome integrity and fertility. Our preliminary data show that loss of GCNA results genomic instability in both Drosophila and C. elegans, indicating conservation of function. The experiments outlined here will provide some of the first insights into the shared mechanisms by which conserved GCNA family members promote genomic integrity and fertility across species. We find that GCNA both interacts with Topoisomerase 2 and prevents the formation of DNA:protein crosslinks (DPCs). GCNA mutant animals display increased chromosome loss, mutation accumulation, and cell cycle defects, placing GCNA at the hub of multiple genome integrity processes. To explore the mechanisms underlying gcna functions in germ cell maintenance, we propose three specific aims: 1) to analyze the etiology of chromosome loss, and to determine the contribution of gcna to cell cycle regulation, 2) to characterize the interaction with topoisomerase 2, identify conserved binding partners, and illuminate the nature and etiology of DPCs, and 3) to understand the domain structure of GCNA and its functional relevance to human fertility.