Functional Characterazation of the H1 Histone Binding Protein, NASP

这项研究项目的长期目标是了解核自身抗原(NASP)的作用 精子蛋白)在基因表达和细胞周期进程中的调节作用。NASP是一个链接器(Hi) 与核小体上的分子复合体中的结合伙伴相互作用的组蛋白伴侣，结合 HSPyo-2和HIT在生精细胞中的表达，并与Kuyo/80和DNA-PK在DNA修复中的作用有关。完毕 在过去的四年里，我们的假设是，在两次有丝分裂过程中，NASP对正常细胞功能都很重要 减数分裂得到了我们的新数据和来自其他几个实验室的数据的充分支持。 我们的数据表明，NASP在细胞周期进程和胚胎发育过程中是必不可少的 Hi组蛋白的发育和NASP的运输和调节可能是表达的关键 由许多基因组成。为了继续我们对NASP的研究，这项建议的具体目标是： 目的#1.这个目标将检验NASP在类固醇受体基因调控中的作用这一假设 通过控制染色质中Hi的水平进行转录。为了检验这一假设，我们将(I)合作 与项目I和项目II一起研究NASP的表达与表达和转录的关系 正常育龄妇女和卵巢癌患者卵巢和子宫内膜类固醇受体活性的研究 多囊卵巢综合征。在这个具体目标的第二部分，我们将(2)确定NASP是否 存在于与Hi组蛋白和DNA结合的染色质核小体复合体中 Hi的翻译后修饰影响其与NASP的结合。具体目标2。这个目标将 检验假设：(I)在精子发生过程中，tNASP与染色质核小体有关 包括HSPyo-2的复合体，HSPyo-2向细胞发出信号，表明它可能从G2过渡到M，以及(2)in 生精细胞α[tNASP-Hi]复合体与Kuyo/KuSo/DNA-PK相关 染色质重塑修复双链DNA断裂并避免细胞凋亡反应 精母细胞和精子细胞。具体目标#3。这个目标将检验NASP表达 在生殖细胞中是精子发生所必需的。为了检验这一假设，我们将生成一个条件 基因敲除小鼠模型，旨在去除NASP在精子发生过程中正常表达的情况。 这项研究将有助于我们对基因调控的理解，特别是有助于 了解多囊卵巢综合征患者不适当的基因表达。

The long-term goals of this research project are to understand the role of NASP (nuclear autoantigenic sperm protein) in the regulation of gene expression and cell cycle progression. NASP is a linker (Hi) histone chaperone that interacts with binding partners in a molecular complex on the nucleosome, binds HSPyo-2 and Hit in spermatogenic cells and associates with Kuyo/8o and DNA-PK in DNA repair. Over the past four years our hypothesis that NASP is important for normal cell function during both mitosis and meiosis has been abundantly supported by our new data and data from several other laboratories. Our data demonstrate that NASP is absolutely required for cell cycle progression and embryonic development and that NASP's transport and regulation of Hi histones may be critical for the expression of a number of genes. To continue our study of NASP, the Specific Aims of this proposal are: Specific aim #1. This aim will test the hypothesis that NASP functions in steroid receptor regulation of gene transcription by controlling the level of Hi in chromatin. To test this hypothesis we will (i) collaborate with Projects I and II to study the expression of NASP in relation to the expression and transcriptional activities of steroid receptors in the ovary and endometrium of normal fertile women and women with polycystic ovarian syndrome. In the second part of this specific aim we will (2) determine whether NASP is present in chromatin nucleosomal complexes bound to Hi histones and DNA and whether post-translational modification of Hi influences its binding to NASP. Specific aim #2. This aim will test the hypothesis that (i) during spermatogenesis tNASP is associated with a chromatin nucleosomal complex that includes HSPyo-2, which signals the cell that it may transition from G2 to M, and that (2) in spermatogenic cells a [tNASP-Hi] complex is associated with Kuyo/KuSo/DNA-PK coincident with chromatin remodeling that repairs double strand DNA breaks and avoids apoptotic responses of spermatocytes and spermatids. Specific aim #3. This aim will test the hypothesis that NASP expression in germ cells is required for spermatogenesis. To test this hypothesis we will generate a conditional knockout mouse model designed to ablate NASP when it is normally expressed during spermatogenesis. This research will contribute to our understanding of gene regulation and in particular will help understand the inappropriate' gene expression in PCOS patients.