HDX-MS Analysis of RORg:RORE:SRC3 Complexes

The retinoic acid receptor-related orphan receptor γ (RORγ) is a ligand-dependent transcription factor that both underpins metabolic and immune functions and provides vantage to manipulate those processes pharmacologically. Despite its importance, our understanding of the ligand-dependent activities of RORγ is far from complete and developing a detailed structural model for RORγ pharmacology could provide a path towards the development of safe and efficacious therapeutics targeting the receptor. Herein, we examine the ligand-dependent assembly of recombinant RORγ:coregulator complexes on cognate DNA response elements using structural proteomics and small angle x-ray scattering. These studies reveal that the RORγ DNA binding domain can bind multiple different sequences of DNA, and that coregulatory proteins may be able to ‘sense’ the ligand- and DNA-bound status of RORγ. Overall, the efforts described herein will illuminate important aspects of RORγ activity and drug development that could lead to more efficacious treatments targeting this important receptor.

视黄酸受体相关孤儿受体γ（retinoic acid receptor-related orphan receptor γ, RORγ）是一类配体依赖性转录因子（ligand-dependent transcription factor），既在代谢与免疫功能中发挥核心支撑作用，也为通过药理学手段调控此类生理过程提供了可行切入点。尽管该受体的生物学功能至关重要，但当前我们对其配体依赖性活性的认知仍远未完备；构建RORγ药理学的精细结构模型，或可为开发靶向该受体的安全有效治疗药物开辟路径。本文借助结构蛋白质组学（structural proteomics）与小角度X射线散射（small angle x-ray scattering）技术，探究了重组RORγ：共调节因子复合物（coregulator complexes）在同源DNA反应元件（cognate DNA response elements）上的配体依赖性组装过程。研究结果显示，RORγ的DNA结合结构域（DNA binding domain）可结合多种不同序列的DNA，且共调节蛋白或能够“感知”RORγ的配体结合与DNA结合状态。总体而言，本文所述的研究工作将阐明RORγ活性与药物开发的关键环节，有望推动针对这一重要受体的更高效治疗方案的研发。