Role of GCNA in preserving genome integrity and fertility

项目总结 我们提出了一种跨物种的方法来理解在胚胎中维持基因组完整性的途径。 细胞。物种的永久存在要求基因组受到保护，不受内源和外源的影响 威胁。在卵子和精子的前体--生殖细胞中，这一点最为关键。产品中的缺陷 警惕途径会导致基因组完整性的丧失，并随后导致后代活力和/或 流产。对一种新发现的高度保守的遗传因子GCNA(生殖细胞核)的详细研究 酸性多肽酶)，将为基因组完整性和生育力的关键决定因素提供新的见解。我们的 初步数据显示，GCNA的缺失导致果蝇和线虫的基因组不稳定， 表示功能上的守恒。这里概述的实验将提供一些关于 保守的GCNA家庭成员促进基因组完整性和生育力的共同机制 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 交叉链接(DPC)。GCNA突变动物表现出更多的染色体丢失、突变积累和 细胞周期缺陷，将GCNA置于多个基因组完整性过程的中心。要探索 Gcna在生殖细胞维持中的作用机制，我们提出了三个具体目标：1) 分析染色体丢失的病因，确定gcna在细胞周期调控中的作用，2) 为了表征与拓扑异构酶2的相互作用，确定保守的结合配对，并阐明 DPC的性质和病因学，以及3)了解GCNA的结构及其功能相关性 人类的生育能力。

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.