Small-molecule Ro-08-2750 interacts with many RNA-binding proteins and elicits MUSASHI2-independent phenotypes.

Kathryn Walters, Marcin Piotr Sajek, Elisabeth Murphy, Aaron Issaian, Amber Baldwin, Evan Harrison, Miles Daniels, Julie A Reisz, Kirk Hansen, Angelo D'Alessandro, Neelanjan Mukherjee

RNA (New York, N.Y.) 2023 Oct

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RNA-binding proteins (RBPs) are key regulators of gene expression. Small molecules targeting these RBP-RNA interactions are a rapidly emerging class of therapeutics for treating a variety of diseases. Ro-08-2750 (Ro) is a small molecule identified as a competitive inhibitor of Musashi (MSI)-RNA interactions. Here, we show that multiple Ro-dependent cellular phenotypes, specifically adrenocortical steroid production and cell viability, are Musashi-2 (MSI2)-independent. Using an unbiased proteome-wide approach, we discovered Ro broadly interacts with RBPs, many containing RRM domains. To confirm this finding, we leveraged the large-scale ENCODE data to identify a subset of RBPs whose depletion phenocopies Ro inhibition, indicating Ro is a promiscuous inhibitor of multiple RBPs. Consistent with broad disruption of ribonucleoprotein complexes, Ro treatment leads to stress granule formation. This strategy represents a generalizable framework for validating the specificity and identifying targets of RBP inhibitors in a cellular context. © 2023 Walters et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Citation

Kathryn Walters, Marcin Piotr Sajek, Elisabeth Murphy, Aaron Issaian, Amber Baldwin, Evan Harrison, Miles Daniels, Julie A Reisz, Kirk Hansen, Angelo D'Alessandro, Neelanjan Mukherjee. Small-molecule Ro-08-2750 interacts with many RNA-binding proteins and elicits MUSASHI2-independent phenotypes. RNA (New York, N.Y.). 2023 Oct;29(10):1458-1470