TY - JOUR
T1 - Global Discovery of Covalent Modulators of Ribonucleoprotein Granules
AU - Ciancone, Anthony M.
AU - Seo, Kyung W.
AU - Chen, Miaomiao
AU - Borne, Adam L.
AU - Libby, Adam H.
AU - Bai, Dina L.
AU - Kleiner, Ralph E.
AU - Hsu, Ku Lung
N1 - Funding Information:
We thank Mark Ross and all members of the Hsu Lab for helpful discussions and review of the manuscript. This work was supported by the National Institutes of Health Grants (DA043571 and GM144472 to K.-L.H.; CA009109 to A.L.B.; and GM132189 to R.E.K.), University of Virginia Cancer Center (NCI Cancer Center Support Grant No. 5P30CA044579-27 to K.-L.H.), the Robbins Family MRA Young Investigator Award from the Melanoma Research Alliance ( http://doi.org/10.48050/pc.gr.80540 to K.-L.H.), the Mark Foundation for Cancer Research (Emerging Leader Award to K.-L.H.), and a Recruitment of Rising Stars Award from CPRIT (RR220063 to K.-L.H.). K.W.S. and R.E.K. thank Princeton University for financial support. We acknowledge the Keck Center for Cellular Imaging for the usage of the Leica SP5X microscopy system (PI:AP, NIH-RR025616). We thank Jeffrey Brulet and Kun Yuan for assistance in synthesis of SuTEx compounds and probes. We thank Heung-Sik Hahm and Emmanuel Toroitich for general help with the SuTEx project. A special thanks to Timothy B. Ware for helping to blind the microscopy slides. We thank D. W. Sanders and C. Brangwynne for providing the G3BP1/2 KO cell line.
Funding Information:
We thank Mark Ross and all members of the Hsu Lab for helpful discussions and review of the manuscript. This work was supported by the National Institutes of Health Grants (DA043571 and GM144472 to K.-L.H.; CA009109 to A.L.B.; and GM132189 to R.E.K.), University of Virginia Cancer Center (NCI Cancer Center Support Grant No. 5P30CA044579-27 to K.-L.H.), the Robbins Family MRA Young Investigator Award from the Melanoma Research Alliance (http://doi.org/10.48050/pc.gr.80540 to K.-L.H.), the Mark Foundation for Cancer Research (Emerging Leader Award to K.-L.H.), and a Recruitment of Rising Stars Award from CPRIT (RR220063 to K.-L.H.). K.W.S. and R.E.K. thank Princeton University for financial support. We acknowledge the Keck Center for Cellular Imaging for the usage of the Leica SP5X microscopy system (PI:AP, NIH-RR025616). We thank Jeffrey Brulet and Kun Yuan for assistance in synthesis of SuTEx compounds and probes. We thank Heung-Sik Hahm and Emmanuel Toroitich for general help with the SuTEx project. A special thanks to Timothy B. Ware for helping to blind the microscopy slides. We thank D. W. Sanders and C. Brangwynne for providing the G3BP1/2 KO cell line.
Publisher Copyright:
© 2023 American Chemical Society.
PY - 2023/5/24
Y1 - 2023/5/24
N2 - 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.
AB - 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.
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U2 - 10.1021/jacs.3c00165
DO - 10.1021/jacs.3c00165
M3 - Article
C2 - 37159397
AN - SCOPUS:85160007779
SN - 0002-7863
VL - 145
SP - 11056
EP - 11066
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 20
ER -