REGULATION OF MEIOSIS IN SPERMATOGENESIS

精子发生中减数分裂的调节

• 批准号: 6085402
• 负责人: Steven Alexander Wasserman
• 金额: $ 21.56万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6085402
• 关键词: Drosophilidae; RNA binding protein; cell cycle proteins; cell differentiation; developmental genetics; enzyme activity; enzyme induction /repression; gene deletion mutation; genetic regulatory element; genetically modified animals; laboratory rabbit; male; meiosis; phosphoprotein phosphatase; point mutation; posttranslational modifications; protein structure function; site directed mutagenesis; sperm; spermatogenesis; testis; yeast two hybrid system

The long term focus of this research is the regulation of cell division in the male germline and the coordination of the cell cycle with spermatocyte development and differentiation. In Drosophila, molecular genetic analyses have identified the boule gene as a key regulatory of meiosis in males. Spermatogenesis in boule mutants is normal up to meiosis, but the G2/M transition of meiosis I fails to occur and males are sterile. Molecular studies have revealed that boule belongs to a conserved family of fertility factors that includes the Daz (Deleted in Azoospermia) gene, a candidate locus for the human Azoospermia Factor (AZF). Whereas the molecular role of its vertebrate homologs in spermatogenesis has not been defined, Boule has been shown to be essential for translation of the cdc25 phosphatase Twine, a key component of the cell cycle machinery. The four specific aims of this proposal are directed at identifying the targets, functional domains, pairing partners, and regulators of the Boule protein. A transgenic approach will be used to analyze the function of Boule, an RNA-binding protein, in Twine regulation. Deletion derivatives and site-directed points mutants will be employed to analyze Boule structure and function both in vitro and in vivo. A yeast two-hybrid screen will be carried out to identify proteins that interact directly with the Boule protein. Lastly, a collection of male-sterile mutants will be screened for regulators for Boule expression and activity. These studies should provide insight into the basic regulatory mechanisms governing cell division in gametogenesis, as well as be of substantial relevance to the investigation and treatment of a significant genetic cause of infertility in men.

这项研究的长期重点是男性种系中细胞分裂的调节以及细胞周期与精子细胞发育和分化的协调。在果蝇中，分子遗传分析已将Boule基因确定为男性减数分裂的关键调节。 Boule突变体中的精子发生是正常的减数分裂，但是我没有发生的减数分裂的G2/M过渡，雄性是无菌的。分子研究表明，Boule属于一个保守的生育因子家族，其中包括DAZ（Azoospermia中删除）基因，这是人类Azoospermia因子（AZF）的候选基因座。尽管尚未定义其脊椎动物同源物在精子发生中的分子作用，但已证明Boule对于CDC25磷酸酶细胞的翻译至关重要，Cdc25磷酸酶麻木是细胞循环机械的关键成分。该提案的四个特定目的旨在识别Boule蛋白质的目标，功能域，配对伙伴和调节剂。一种转基因方法将用于分析在链条调节中BOULE（RNA结合蛋白）的功能。将使用缺失衍生物和定点点突变体来分析体外和体内的Boule结构和功能。将进行酵母两杂交筛选，以鉴定直接与Boule蛋白相互作用的蛋白质。最后，将筛选一组男性突变体的集合，以进行调节剂的表达和活性。这些研究应洞悉有关配子发生中细胞分裂的基本调节机制，并与研究和治疗男性不孕的重大遗传原因有关。