Functions of the Tudor-domain containing protein TDRD6 in male germ cells

含有 Tudor 结构域的蛋白 TDRD6 在雄性生殖细胞中的功能

• 批准号: 186884645
• 负责人: Professor Dr. Rolf Jessberger
• 金额: --
• 依托单位: Institut für Physiologische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/186884645?language=en
• 关键词: Functions; Tudor; domain; containing; protein

Tudor domains bind methylated arginine in proteins and may bind nucleic acids. In germ cells, the molecular and cellular roles of several tudor domain bearing proteins are little understood, yet some reports indicate central and essential functions. TDRD6 is specifically expressed starting in late pachytene in spermatocytes. Initially distributed throughout the cell it concentrates during meiosis in some dispersed granule-like chromatoid bodies (CB; type I), then in spermatids it concentrates in the one perinuclear CB (type II). In the 1st funding period we identified and characterized two hitherto unknown major functions for TDRD6, both relating to large protein-RNA complexes: roles (i) in nonsense-mediated mRNA decay (NMD) and (ii) in spliceosome assembly. Our studies on TDRD6 in spermatid NMD were recently published (Fanourgakis et al., 2016), the data on the role of TDRD6 in spermatocyte splicing are submitted (Akpinar et al., 2016). Very recent data also suggest an additional nuclear localisation of TDRD6. Based on the data generated during the 1st funding period our hypothesis is that TDRD6 acts in assembling diverse RNA-related protein complexes and intracellular bodies that harbor arginine-methylated proteins. In addition, TDRD6 controls arginine methylation itself by regulating the methyl transferase PRMT5. Therefore, TDRD6 appears as a central regulator of processes in spermatogenesis, which depend on symmetric dimethylation of arginine of proteins.In the requested 2nd funding period, we plan to determine fundamental properties of TDRD6 underlying its role in protein complex assembly and its relationship to PRMT5.Our specific aims for the next funding period therefore are:(1) To understand the regulation of TDRD6 (2) To understand how TDRD6 controls PRMT5(3) To decipher the role of TDRD6 in the nucleusWe have developed a plethora of tools and methods to address these questions including TDRD6 deficiency mice, mice carrying a fully functional TDRD6-LAP transgene, mice allowing sorting of specific spermatocyte/spermatid populations, as well as RIP, chromatoid body proteomics and transcriptomics. We expect to obtain important insights into TDRD6 and PRMT5, which will in mid-term allow us to further study in detail the activities of TDRD6 complexes in NMD and splicing as well as to identify potential further roles of TDRD6.

Tudor结构域结合蛋白质中的甲基化精氨酸并且可以结合核酸。在生殖细胞中，几个都铎结构域轴承蛋白的分子和细胞的作用是很少了解，但一些报告表明，中央和必要的功能。TDRD 6在精母细胞的粗线期晚期开始特异性表达。最初分布在整个细胞中，在减数分裂期间集中在一些分散的颗粒状拟染色体体（CB; I型）中，然后在精子细胞中集中在一个核周CB（II型）中。在第一个资助期，我们确定并表征了TDRD 6的两个迄今未知的主要功能，这两个功能都与大蛋白质-RNA复合物有关：（i）无义介导的mRNA衰变（NMD）和（ii）剪接体组装中的作用。我们对精子细胞NMD中TDRD 6的研究最近发表（Fanourgakis et al.，2016），提交了关于TDRD 6在精母细胞剪接中的作用的数据（Akpinar等人，2016年）。最近的数据也表明TDRD 6的额外核定位。基于第一个资助期产生的数据，我们的假设是TDRD 6在组装不同的RNA相关蛋白质复合物和含有精氨酸甲基化蛋白质的细胞内体中起作用。此外，TDRD 6通过调节甲基转移酶PRMT 5来控制精氨酸甲基化本身。因此，TDRD 6似乎是精子发生过程中的一个中心调节因子，而精子发生过程依赖于蛋白质精氨酸的对称二甲基化。在申请的第二个资助期，我们计划确定TDRD 6在蛋白质复合物组装中的基本特性及其与PRMT 5的关系。因此，我们下一个资助期的具体目标是：（1）了解TDRD 6的调节（2）了解TDRD 6如何控制PRMT 5（3）为了破译TDRD 6在细胞核中的作用，我们已经开发了大量的工具和方法来解决这些问题，包括TDRD 6缺陷小鼠，携带完全功能性TDRD 6-TRY 1转基因的小鼠，允许分选特定精母细胞/精子细胞群体的小鼠，以及RIP、拟染色体体蛋白质组学和转录组学。我们期望获得对TDRD 6和PRMT 5的重要见解，这将使我们能够在中期进一步详细研究TDRD 6复合物在NMD和剪接中的活性，以及确定TDRD 6的潜在进一步作用。