Functional Characterazation of the H1 Histone Binding Protein, NASP

H1 组蛋白结合蛋白 (NASP) 的功能表征

• 批准号: 7315897
• 负责人: MICHAEL GENE ORAND
• 金额: $ 30.11万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7315897
• 关键词: Address; Apoptotic; Binding; Cell Cycle; Cell Cycle Progression; Cell Nucleus; Cell physiology; Chromatin; Complement; Complex; DNA; DNA Double Strand Break; DNA Repair; DNA biosynthesis; DNA chemical synthesis; DNA-dependent protein kinase; Data; Embryo; Embryonic Development; Endometrium; Equilibrium; Estrogen Receptors; G2 Phase; G22P1 gene; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Germ Cells; Goals; Grant; Histone H1; Histone H1(s); Histones; In Vitro; Knock-out; Laboratories; Meiosis; Mitosis; Molecular; Molecular Chaperones; Names; Nonhomologous DNA End Joining; Normal Cell; Nuclear; Nucleosomes; Numbers; Ovarian; Ovary; Patients; Phase Transition; Play; Post-Translational Protein Processing; Regulation; Reporter Genes; Research; Research Project Grants; Role; Signal Transduction; Somatic Cell; Spermatids; Spermatocytes; Spermatogenesis; Spermatogenic Cell; Staging; Steroid Receptors; Syndrome; System; Testing; Testis; Type I DNA Topoisomerases; Woman; XRCC5 gene; chromatin remodeling; histone-binding proteins; model design; mouse model; null mutation; prevent; protein expression; protein function; repaired; response; sperm cell; sperm protein

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.

本研究项目的长期目标是了解NASP（核自身抗原）的作用， 精子蛋白）在基因表达和细胞周期进程的调节中的作用。NASP是接头（Hi） 组蛋白伴侣蛋白与核小体上的分子复合物中的结合伴侣相互作用， HSPyo-2和Hit在生精细胞中的表达，并与DNA修复中的Kuyo/8o和DNA-PK相关。超过 在过去的四年里，我们假设NASP在有丝分裂和有丝分裂过程中对正常细胞功能都很重要， 我们的新数据和其他几个实验室的数据充分支持了减数分裂。 我们的数据表明，NASP是细胞周期进程和胚胎发育所必需的。 NASP对Hi组蛋白的转运和调节可能对H1组蛋白的表达至关重要。 一系列的基因。为了继续我们对NASP的研究，本提案的具体目标是： 目标#1。这一目的将验证NASP在类固醇受体基因调控中的作用的假设。 通过控制染色质中Hi的水平来调控转录。为了验证这一假设，我们将（i）合作 与项目I和II一起研究NASP的表达与表达和转录的关系， 正常生育妇女和不孕妇女卵巢和子宫内膜类固醇受体活性的测定 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物中， Hi的翻译后修饰影响其与NASP的结合。具体目标#2这一目标将 检验以下假设：（i）在精子发生过程中，tNASP与染色质核小体相关 复合物，包括HSPyo-2，其信号细胞，它可以从G2过渡到M，和（2）在 生精细胞a [tNASP-Hi]复合物与Kuyo/KuSo/DNA-PK相关联， 染色质重塑修复双链DNA断裂，避免细胞凋亡反应， 精母细胞和精子细胞。具体目标#3这一目的将检验NASP表达 是精子发生所必需的。为了验证这个假设，我们将生成一个条件 基因敲除小鼠模型，其设计为当NASP在精子发生期间正常表达时消融NASP。 这项研究将有助于我们理解基因调控，特别是将有助于 了解PCOS患者中不适当的基因表达。