The Fe(II and 2OG dependent dioxygenase Jmjd6 in Hydra: Highly conserved in all animals, essential in vertebrates - does it have an evolutionary conserved function

水螅中的 Fe(II 和 2OG 依赖性双加氧酶 Jmjd6：在所有动物中高度保守，在脊椎动物中必不可少 - 它是否具有进化保守功能

• 批准号: 471245357
• 负责人: Professorin Dr. Angelika Böttger
• 金额: --
• 依托单位: Abteilung für Zell- und Entwicklungsbiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/471245357?language=en
• 关键词: Fe; II; 2OG; dependent; dioxygenase

The JmjC-domain family of Fe(II) and 2-oxoglutarate dependent dioxygenases contains about 60 members in humans. They are involved in hydroxylating substrates such as proteins or nucleic acids; and in reversing arginine or lysine N-methylations dividing them in hydroxylases and demethylases. As their activity depends on oxygen tension and tricarboxylic acid intermediates they are able to link protein and nucleic acid modifications with environmental conditions and this is important for some dynamic epigenetic modifications. Jmjd6 is a member of this family with great biological importance for vertebrate embryonic development and oncogenic mechanisms. Despite the fact that Jmjd6 is essential for mouse and zebrafish development the molecular function of human Jmjd6 is insufficiently understood and in parts controversially discussed. In early metazoans there is almost no information about the biochemical and physiological roles of Jmjd6. The remarkably strong sequence conservation of Jmjd6 from Hydra to humans indicates that there is a conserved function that has to be uncovered. We therefore suggest to characterize Jmjd6 in the early metazoan Hydra, which is an ideal model organism to combine biochemical with developmental studies. Using mouse monoclonal antibodies against Hydra-Jmjd6 we will identify proteins and protein domains that interact with Hydra-Jmjd6 and compare those with the human Jmjd6 interactome. We will carry out X-ray structural analysis to understand the evolution of the 3-D-structure of Jmjd6 and define its enzymatic activity towards the identified substrates. In addition, we are planning to study the effects of hypoxia and nutrition on mRNA- and protein levels as well as on the subnuclear distribution of Jmjd6, and we will analyze the role of Jmjd6 for Hydra developmental processes such as regeneration and budding. Characterization of transgenic lines and application of pan-specific Fe(II)- and 2OG dependent dioxygenase inhibitors, Jmjd6 specific inhibitors and siRNAs will be employed. A second knockdown approach generating Jmjd6 knockdown Hydra-lines using the vector pHyVec12 will complement this. We expect that these investigations will shed light on the molecular and biological roles of Jmjd6 in humans.

铁（II）和2-氧葡萄糖酸盐依赖性双加氧酶的jmjc结构域家族在人类中包含约60个成员。它们参与羟基化底物，如蛋白质或核酸；并逆转精氨酸或赖氨酸的n -甲基化，使它们在羟化酶和去甲基化酶中分裂。由于它们的活性取决于氧张力和三羧酸中间体，它们能够将蛋白质和核酸修饰与环境条件联系起来，这对于一些动态表观遗传修饰是重要的。Jmjd6是该家族的成员，在脊椎动物胚胎发育和致癌机制中具有重要的生物学意义。尽管Jmjd6对小鼠和斑马鱼的发育至关重要，但人类Jmjd6的分子功能还没有得到充分的了解，并且在某些方面存在争议。在早期后生动物中，几乎没有关于Jmjd6的生化和生理作用的信息。从Hydra到人类，Jmjd6的序列保守性非常强，这表明有一个保守的功能必须被揭示。因此，我们建议在早期后生动物水螅中对Jmjd6进行表征，水螅是将生化与发育研究结合起来的理想模式生物。利用小鼠抗Hydra-Jmjd6单克隆抗体，我们将鉴定与Hydra-Jmjd6相互作用的蛋白和蛋白结构域，并与人Jmjd6相互作用组进行比较。我们将进行x射线结构分析，了解Jmjd6的三维结构演变，并确定其对鉴定底物的酶活性。此外，我们计划研究缺氧和营养对Jmjd6 mRNA和蛋白水平的影响以及对Jmjd6亚核分布的影响，并分析Jmjd6在水螅再生和出芽等发育过程中的作用。转基因品系的鉴定以及泛特异性的Fe(II)-和2OG依赖性双加氧酶抑制剂、Jmjd6特异性抑制剂和sirna的应用将被采用。第二种knockdown方法使用pHyVec12向量生成Jmjd6 knockdown Hydra-lines，将补充这一点。我们期望这些研究将揭示Jmjd6在人类中的分子和生物学作用。