The role of the Dead end protein in controlling germ cell fate

死端蛋白在控制生殖细胞命运中的作用

• 批准号: 388866775
• 负责人: Professor Dr. Erez Raz
• 金额: --
• 依托单位: Institut für Zellbiologie
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/388866775?language=en
• 关键词: role; Dead; end; protein; controlling

The migration, specification and fate maintenance of germ cells is a conserved at the cellular and molecular levels. A key molecule that is required during different steps of germline development is the RNA-binding protein Dead end.In the absence of the Dead end protein, the germline is lost giving rise to sterile animals. We could show that Dead end functions in germ cells to protect their unipotent differentiation potential by inhibiting somatic differentiation such that gametes can be formed. Global analysis of the effect of Dead end on the transcriptome of germ cells revealed that a range of somatic differentiation factors become expressed in Dead end depleted cells, concomitantly with a reduction in the transcription level of RNAs linked to germline development. Importantly, among the genes suppressed by Dead end are genes encoding for pluripotency factors such as Nanog and the Oct4 homolog.In the next funding period we propose to expand the scope of understanding the function of Dead end to later stages of germline development. The proposed work first involves the establishment of an experimental system for inhibiting Dead end at different stages. This procedure relies on inducing Dead end protein degradation by expressing specific nanobodies directed against the tagged protein. The transcriptome and chromatin structure of germ cells knocked down at different stages of their development will be assayed to reveal the role of the Dead end at stages previously not explored regarding this issue. The role of the regulation of pluripotency proteins by Dead end will be studied in experiments where Dead end function is inhibited in combination with manipulations (genetic mutations of and over-expression) in pluripotency-promoting genes. Together, the results of these studies will enhance the understanding of the molecular mechanisms that direct the developmental potential of the germline to the formation of gametes.

生殖细胞的迁移、特化和命运维持在细胞和分子水平上是保守的。RNA结合蛋白Dead end是生殖细胞发育过程中的一个关键分子。如果没有Dead end蛋白，生殖细胞就会丢失，从而导致不育动物。我们可以证明死端在生殖细胞中的功能是通过抑制体细胞分化来保护它们的单能分化潜能，从而可以形成配子。死端对生殖细胞转录组的影响的全球分析显示，一系列体细胞分化因子在死端耗尽的细胞中表达，伴随着与生殖系发育相关的RNA转录水平的降低。重要的是，在被Dead end抑制的基因中，有编码多能性因子的基因，如Nanog和Oct4同系物。在下一个资助期，我们建议将Dead end功能的理解范围扩大到生殖系发育的后期阶段。本文首先建立了一个在不同阶段抑制死胡同的实验系统。该程序依赖于通过表达针对标记蛋白质的特异性纳米抗体来诱导死端蛋白质降解。将测定在其发育的不同阶段被敲除的生殖细胞的转录组和染色质结构，以揭示死端在以前未探讨过的阶段中的作用。通过死端调节多能性蛋白的作用将在实验中研究，其中死端功能被抑制，并结合多能性促进基因的操作（基因突变和过表达）。总之，这些研究的结果将增强对指导生殖系发育潜力形成配子的分子机制的理解。