Characterization of a Testis Kinase Family (Tssk)

睾丸激酶家族 (Tssk) 的表征

• 批准号: 7093986
• 负责人: Pablo E. Visconti
• 金额: $ 7.85万
• 依托单位: UNIVERSITY OF MASSACHUSETTS AMHERST
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7093986
• 关键词: DNA binding protein; antibody; biological signal transduction; cell differentiation; cell line; developmental genetics; enzyme activity; enzyme substrate; heat shock proteins; histones; immunoprecipitation; laboratory mouse; meiosis; phosphorus; phosphorylation; protein isoforms; protein localization; protein protein interaction; serine threonine protein kinase; sperm; spermatogenesis; testis; two dimensional gel electrophoresis

DESCRIPTION (provided by applicant): Post translational modifications of proteins through phosphorylation play a role in many cellular processes such as the transduction of extracellular signals, intracellular transport, and cell cycle progression. Our group is interested in the study of signaling events that are involved in sperm differentiation and function. Although meiotic events are well characterized in the female germ cells, the regulation of male germ cell meiosis and differentiation into mature sperm is still not well understood. Recently, a novel family of serine/threonine kinases (testes specific serine kinases (Tssk)) has been found post meiotically expressed in the male germ cells. We have demonstrated the presence of 1or more members of the Tssk family in mature mammalian sperm (Hao et al., 2004) (and PR) and in addition, we have obtained evidence that Tssk homologues are expressed in the testes of invertebrates. The conserved testicular expression pattern of Tssk genes, as well as the importance of phosphorylation in signaling processes, strongly suggests that Tssk(s) have important role(s) in germ cell differentiation and/or sperm function. This proposal is designed to set the grounds to understand the function(s) of Tssk(s) in germ cells and in mature sperm. Future directions of this project include the analysis of null mutants (KO) of Tssk(s), as well as the investigation of signaling pathways involving Tssk(s) using genetic methods available in invertebrate models. Toward these goals, it will be necessary first to create tools for these studies and to characterize these kinases. Taking this into consideration, this application will focus on: 1) The generation of specific antibodies against the 5 members of the Tssk kinase family; 2) The identification of the Tssk(s) present in mature sperm and the investigation of their subcellular localization; 3) The biochemical characterization of Tssk4 kinase activity and the study of potential interacting partners. Although not part of this application, in collaboration with Dr. Mitch Eddy from NTH, a mouse KO of Tssk4 is in the process of being produced. Importantly, the proposed research will be useful to begin to illuminate the pathways of Tssk gene function. The predicted importance of Tssks in spermiogenesis and/or sperm function has been recently confirmed by phenotypic analysis of Tssk6 KOs (also known as Sstk)(Spiridonov et al., 2005); these null mutant mice present a sterile phenotype in the males but not in the females. The tools generated as part of this application will be very important for the analysis of Tssk KO phenotypes and to start the study of Tssk function. The predicted importance of Tssks in spermiogenesis and/or sperm function suggests that this research can ultimately help to design novel human contraceptives.

描述（由申请人提供）：通过磷酸化进行的蛋白质翻译后修饰在许多细胞过程中发挥作用，例如细胞外信号转导、细胞内转运和细胞周期进程。我们的团队对研究精子分化和功能中涉及的信号事件感兴趣。虽然减数分裂事件在雌性生殖细胞中得到了很好的表征，但雄性生殖细胞减数分裂和分化为成熟精子的调控仍然没有得到很好的理解。最近，一个新的丝氨酸/苏氨酸激酶家族（睾丸特异性丝氨酸激酶（Tssk））被发现在减数分裂后的雄性生殖细胞中表达。我们已经证明了在成熟的哺乳动物精子中存在1个或多个Tssk家族成员（Hao等人，2004）（和PR），此外，我们已经获得的证据表明，Tssk同源物在无脊椎动物的睾丸中表达。Tssk基因的保守睾丸表达模式以及磷酸化在信号传导过程中的重要性强烈表明Tssk在生殖细胞分化和/或精子功能中具有重要作用。该提案旨在为了解Tssk在生殖细胞和成熟精子中的功能奠定基础。该项目的未来方向包括分析Tssk（s）的无效突变体（KO），以及使用无脊椎动物模型中可用的遗传方法研究涉及Tssk（s）的信号通路。为了实现这些目标，首先有必要为这些研究创建工具并表征这些激酶。考虑到这一点，本申请将集中于：1）产生针对Tssk激酶家族5个成员的特异性抗体; 2）鉴定成熟精子中存在的Tssk及其亚细胞定位的研究; 3）Tssk 4激酶活性的生化表征和潜在相互作用伴侣的研究。虽然不是本申请的一部分，但与NTH的Mitch Eddy博士合作，正在生产Tssk 4的小鼠KO。重要的是，拟议的研究将有助于开始阐明Tssk基因功能的途径。Tssk在精子发生和/或精子功能中的预测重要性最近已经通过Tssk 6科斯（也称为Sstk）的表型分析得到证实（Spiridonov等人，2005）;这些无效突变小鼠在雄性中而不是在雌性中呈现不育表型。作为该应用程序的一部分生成的工具对于分析Tssk KO表型和开始Tssk功能的研究非常重要。Tssks在精子发生和/或精子功能中的预测重要性表明，这项研究最终有助于设计新型人类避孕药。