SPERM CHROMATIN PROTEOMICS

精子染色质蛋白质组学

• 批准号: 7957805
• 负责人: BARBARA J MEYER
• 金额: $ 0.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957805
• 关键词: Aneuploidy; Biology; Caenorhabditis elegans; Cataloging; Catalogs; Chromatin; Chromatin Structure; Chromosome Segregation; Chromosomes; Clinical; Computer Retrieval of Information on Scientific Projects Database; Coupled; Cytology; DNA; F Factor; Failure; Fertility; Fertilization in Vitro; Funding; Fungal Genome; Goals; Grant; Infertility; Institution; Male Contraceptive Agents; Male Infertility; Male Sterility; Mammals; Medical; Meiosis; Mus; Nematoda; Oogenesis; Process; Proteins; Proteomics; RNA Interference; Research; Research Personnel; Resources; Role; Source; Spermatogenesis; Technology; United States National Institutes of Health; base; cell type; gene function; knockout gene; male; reproductive; sperm cell; sperm protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Male infertility is a long-standing enigma of significant medical concern. The integrity of sperm chromatin is a clinical indicator of male fertility and in vitro fertilization potential: chromosome aneuploidy and DNA decondensation or damage are correlated with reproductive failure. Identifying conserved proteins important for sperm chromatin structure and packaging can reveal universal causes of infertility. Here we combine proteomics, cytology and functional analysis in Caenorhabditis elegans to identify spermatogenic chromatin-associated proteins that are important for fertility. Our strategy employed multiple steps: purification of chromatin from comparable meiotic cell types, namely those undergoing spermatogenesis or oogenesis; proteomic analysis by multidimensional protein identification technology (MudPIT) of factors that co-purify with chromatin; prioritization of sperm proteins based on abundance; and subtraction of common proteins to eliminate general chromatin and meiotic factors. Our approach reduced 1,099 proteins co-purified with spermatogenic chromatin, currently the most extensive catalogue, to 132 proteins for functional analysis. Reduction of gene function through RNA interference coupled with protein localization studies revealed conserved spermatogenesis-specific proteins vital for DNA compaction, chromosome segregation, and fertility. Unexpected roles in spermatogenesis were also detected for factors involved in other processes. Our strategy to find fertility factors conserved from C. elegans to mammals achieved its goal: of mouse gene knockouts corresponding to nematode proteins, 37% (7/19) cause male sterility. Our list therefore provides significant opportunity to identify causes of male infertility and targets for male contraceptives.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 男性不育症是长期以来的重要医学关注的谜团。精子染色质的完整性是雄性生育能力和体外受精潜力的临床指标：染色体非整倍性和DNA脱气或损伤与生殖衰竭相关。确定对精子染色质结构和包装重要的保守蛋白质可以揭示不孕的普遍原因。在这里，我们结合了秀丽隐杆线虫中的蛋白质组学，细胞学和功能分析，以鉴定出对生育至关重要的精子染色质相关蛋白。我们的策略采用了多个步骤：从可比的减数分裂细胞类型中纯化染色质，即接受精子发生或卵子发生的染色质；通过多维蛋白质鉴定技术（MUDPIT）与染色质共纯化的因素进行蛋白质组学分析；基于丰度的精子蛋白的优先级；和减去普通蛋白以消除一般染色质和减数分裂因子。我们的方法将1,099种蛋白质与精子染色质（目前是最广泛的目录）共纯化为132种蛋白质，以进行功能分析。通过RNA干扰以及蛋白质定位研究的降低基因功能表明，对DNA压实，染色体分离和生育力至关重要的精子发生特异性蛋白质。还检测到其他过程中涉及的因素，还检测到了精子发生中的意外作用。我们发现从秀丽隐杆线虫到哺乳动物保守的生育因子的策略实现了其目标：与线虫蛋白相对应的小鼠基因敲除，37％（7/19）导致男性无菌。因此，我们的列表提供了巨大的机会来确定男性不育症的原因和男性避孕药的目标。