Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States.
Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
J Am Chem Soc. 2023 May 24;145(20):11056-11066. doi: 10.1021/jacs.3c00165. Epub 2023 May 9.
Stress granules (SGs) and processing-bodies (PBs, P-bodies) are ubiquitous and widely studied ribonucleoprotein (RNP) granules involved in cellular stress response, viral infection, and the tumor microenvironment. While proteomic and transcriptomic investigations of SGs and PBs have provided insights into molecular composition, chemical tools to probe and modulate RNP granules remain lacking. Herein, we combine an immunofluorescence (IF)-based phenotypic screen with chemoproteomics to identify sulfonyl-triazoles (SuTEx) capable of preventing or inducing SG and PB formation through liganding of tyrosine (Tyr) and lysine (Lys) sites in stressed cells. Liganded sites were enriched for RNA-binding and protein-protein interaction (PPI) domains, including several sites found in RNP granule-forming proteins. Among these, we functionally validate G3BP1 Y40, located in the NTF2 dimerization domain, as a ligandable site that can disrupt arsenite-induced SG formation in cells. In summary, we present a chemical strategy for the systematic discovery of condensate-modulating covalent small molecules.
应激颗粒（SGs）和处理体（PBs，P 体）是普遍存在且广泛研究的核糖核蛋白（RNP）颗粒，参与细胞应激反应、病毒感染和肿瘤微环境。虽然对 SGs 和 PBs 的蛋白质组学和转录组学研究提供了对分子组成的深入了解，但用于探测和调节 RNP 颗粒的化学工具仍然缺乏。在此，我们结合基于免疫荧光（IF）的表型筛选和化学蛋白质组学，鉴定出能够通过结合应激细胞中酪氨酸（Tyr）和赖氨酸（Lys）位点来预防或诱导 SG 和 PB 形成的磺酰三唑（SuTEx）。结合位点富含 RNA 结合和蛋白质-蛋白质相互作用（PPI）结构域，包括在形成 RNP 颗粒的蛋白质中发现的几个位点。在这些位点中，我们验证了位于 NTF2 二聚化结构域中的 G3BP1 Y40 是一个可配体的位点，可破坏亚砷酸盐诱导的细胞中 SG 的形成。总之，我们提出了一种用于系统发现凝聚物调节共价小分子的化学策略。
