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

Fe（II）和2-酮戊二酸依赖性双加氧酶的JmjC结构域家族在人类中包含约60个成员。它们参与羟化底物如蛋白质或核酸;并参与逆转精氨酸或赖氨酸N-甲基化，将其分为羟化酶和脱甲基酶。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很重要。jmjd 6是该家族的一员，对脊椎动物胚胎发育和致癌机制具有重要的生物学意义。尽管Jmjd 6对小鼠和斑马鱼的发育至关重要，但人类Jmjd 6的分子功能还没有得到充分的理解，并且部分地进行了有争议的讨论。在早期后生动物中，几乎没有关于Jmjd 6的生化和生理作用的信息。从水螅到人类的Jmjd 6序列非常强的保守性表明，有一个保守的功能，必须被发现。因此，我们建议在早期后生动物水螅，这是一个理想的模式生物联合收割机生化与发育研究的特点Jmjd 6。使用针对Hydra-Jmjd 6的小鼠单克隆抗体，我们将鉴定与Hydra-Jmjd 6相互作用的蛋白质和蛋白质结构域，并将其与人类Jmjd 6相互作用组进行比较。我们将进行X射线结构分析，以了解Jmjd 6的3-D结构的演变，并确定其对已鉴定底物的酶活性。此外，我们正计划研究缺氧和营养对mRNA和蛋白质水平以及Jmjd 6亚核分布的影响，我们将分析Jmjd 6在水螅发育过程中的作用，如再生和出芽。将采用转基因系的表征和泛特异性Fe（II）和2 OG依赖性双加氧酶抑制剂、Jmjd 6特异性抑制剂和siRNA的应用。使用载体pHyVec 12产生Jmjd 6敲低Hydra-lines的第二敲低方法将对此进行补充。我们希望这些研究将揭示Jmjd 6在人类中的分子和生物学作用。

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.