Post-translational modifications in RORgt-dependent immune cell functions

家长奖项目总结 转录因子RORgt是淋巴细胞分化所必需的，它不仅保护屏障表面 不受感染，但也对炎症性疾病有重大贡献。使用蛋白质组学方法，我们发现 RORgt在翻译后(PTM)被大量修饰，并与蛋白质和RNA辅助调节因子相互作用。 在Th17细胞中，RORgt被磷酸化。为了评估这些PTM的影响，我们生成了STOKS- 在内源性RORC基因座携带修饰零等位基因的小鼠中。RORgt靶基因表达为 在细胞类型和组织特异性的PTM突变动物中显著减少。更详细的 不同PTM调控RORgt的特性及其在组织特异性RORgt中的作用 互动伙伴可为免疫和免疫环境中的治疗干预提供新方法 自身免疫性疾病。在目标1中，我们将首先确定RORgt的磷酸化是否有助于促进靶向 基因转录水平上的染色质可及性、转录因子染色质占有率，还是变化 在Th17细胞中形成蛋白质伙伴关系。在目标2中，我们将确定磷酸化在RORgt依赖中的作用 多发性硬化症和结肠炎小鼠模型中的淋巴样细胞。在目标3中，我们将描述相互作用 MAPK12/p38g和RORgt之间的关系以确定其在RORgt磷酸化和转录中的作用 Th17细胞的活性。这里提出的实验除了解决特定的机械问题外 概述了每个目标，也将为我们的长期目标奠定基础，了解转录因子是如何 在分子水平上实现细胞类型的特定功能。

Project Summary of Parent Award Transcription factor RORgt is required for the differentiation of lymphoid cells that not only protect barrier surfaces from infection but also contribute significantly to inflammatory diseases. Using proteomics approaches, we found RORgt to be heavily modified post-translationally (PTM) and interacted with both protein and RNA coregulators. In Th17 cells, RORgt is phosphorylated. To evaluate the implication of these PTMs, we have generated knock- in mice carrying modification-null alleles at the endogenous rorc locus. RORgt target gene expressions were significantly reduced in PTM mutant animals in a cell-type and tissue-specific manner. A more detailed characterization of how RORgt is regulated by different PTMs and their contribution to tissue-specific RORgt interaction partners may provide new approaches for therapeutic intervention in the setting of immunity and autoimmune conditions. In Aim 1, we will first determine if phosphorylation of RORgt helps to facilitate target gene transcription at the level of chromatin accessibility, transcription factor chromatin occupancy, or changes to protein partnership in Th17 cells. In Aim 2, we will determine the role of phosphorylation in RORgt-dependent lymphoid cells in mouse models of multiple sclerosis and colitis. In Aim 3, we will characterize the interaction between MAPK12/p38g and RORgt to determine its contribution to RORgt phosphorylation and transcription activity in Th17 cells. The experiments proposed here in addition to addressing specific mechanistic questions outlined in each Aim, will also lay the groundwork for our long-term goal, to understand how transcription factors achieve cell-type specific functions at the molecular level.