Post-translational modifications in RORgt-dependent immune cell functions

项目摘要 Rorγt是3型固有淋巴样细胞(ILC3)和辅助性T细胞(Th17)分化所必需的 不仅保护屏障表面不受感染，而且还极大地促进了炎症性疾病。vbl.使用 蛋白质组学方法，我们发现RoRγt在翻译后被大量修饰，并与 蛋白质和RNA共同调节因子的数量。除了先前已知的磷酸化和乙酰化 ，我们发现了RoRγt的丝氨酸510位的新磷酸化和赖氨酸516位的泛素化。 评估这些PTM的含义，我们已经在以下位置产生了携带修饰零等位基因的敲入小鼠 内源性RORC基因座。在ptm突变动物中，rorγt靶基因表达显著降低。 以特定于细胞类型和组织的方式。更详细地描述RoRγt是如何由这些 PTM及其对组织特异性RoRγt相互作用伙伴的贡献可能提供新的方法 在免疫和自身免疫条件下的治疗干预。对于第一个目标，我们将 确定RoR-γ-tPTMS在IL-3介导的保护性免疫和Th17依赖的自身免疫中的作用 条件。在目标2中，我们将评估rorγt ptm零突变对基因组占有率的贡献。 Rorγt及其转录辅助调节因子、染色质可及性和Th17细胞中的整体转录。在……里面 目的3，在体内鉴定在S510和K516修饰RoRγt所必需的上游酶。

Project Summary RORγt is required for the differentiation of type 3 innate lymphoid cells (ILC3) and T helper 17 cells (Th17) that not only protect barrier surfaces from infection but also contribute significantly to inflammatory diseases. Using proteomics approaches, we found RORγt to be heavily modified post-translationally (PTM) and interact with a number of protein and RNA coregulators. In addition to the known phosphorylation and acetylation previously documented, we found novel phosphorylation at serine 510 and ubiquitination at lysine 516 of RORγt. To evaluate the implication of these PTMs, we have generated knock-in mice carrying modification-null alleles at the endogenous rorc locus. RORγt target gene expressions were significantly reduced in PTM mutant animals in a cell type and tissue specific manner. A more detailed characterization of how RORγt is regulated by these PTMs and their contribution to tissue-specific RORγt interaction partners may provide new approaches for therapeutic intervention in the setting of immunity and autoimmune conditions. For the first aim, we will determine the role of RORγt PTMs in ILC3-mediated protective immunity and Th17-dependent autoimmune conditions. In Aim 2, we will evaluate the contribution of RORγt PTM null mutations to the genomic occupancy of RORγt and its transcription coregulators, chromatin accessibility, and global transcriptions in Th17 cells. In Aim 3, we will identify upstream enzymes essential for modifying RORγt at S510 and K516 in vivo.