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Cited by 6RNA Research and SplicingClick Chemistry and ApplicationsProtein Degradation and Inhibitors

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Structural and mechanistic analysis of covalent ligands targeting the RNA-binding protein NONO

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Lindsey, Garrett L., Hockley, Thomas K., Gomez, Alejandro Villa, Marshall, Andrew C., Brothers, William R., Finney, Colin T., Gross, Jacob, Fox, Archa H., Yeo, Gene W., Melillo, Bruno, Bond, Charles S., Cravatt, Benjamin F.·2026·Cell Chemical Biology
DOI 10.1016/j.chembiol.2025.12.010PMC PMC12918760

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Abstract

RNA-binding proteins (RBPs) play important roles in mRNA transcription, processing, and translation. Chemical tools are lacking for RBPs, which has hindered efforts to perturb and understand RBP function in cells. We previously described a chloroacetamide compound ( R )-SKBG-1 that covalently binds the RBP NONO and stabilizes its interactions with mRNAs, leading to transcriptional remodeling and suppression of cancer cell growth. Here, we report the crystal structure of an ( R )-SKBG-1:NONO complex, which confirms covalent modification of cysteine-145 at a pocket proximal to the RNA-binding interface of the protein. We show that this pocket can also be targeted by a lower reactivity chlorofluoroacetamide analog ( R , R )-GL-373, which retains the pharmacological properties of ( R )-SKBG-1, including blockade of estrogen receptor expression in breast cancer cells, while displaying much greater proteome-wide selectivity. Our findings thus show that NONO can be targeted by covalent ligands with high specificity to pharmacologically suppress pro-tumorigenic gene products in cancer cells.

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Metadata

Authors
Lindsey, G. L., Hockley, T. K., Gomez, A. V., Marshall, A. C., Brothers, W. R., Finney, C. T., Gross, J., Fox, A. H., Yeo, G. W., Melillo, B., Bond, C. S., & Cravatt, B. F.
Affiliations
  • Scripps Research Institute (US)
  • The University of Western Australia (AU)
  • University of California San Diego (US)
  • Vividion Therapeutics (United States) (US)
Corresponding author
  • Bond, Charles S.
  • Cravatt, Benjamin F.
ORCID
Venue
Cell Chemical Biology
Publisher
Elsevier BV
Published
2026-02
Volume
33
Issue
2
Pages
256-267.e11
ISSN
  • 2451-9448 (ISSN-L)
  • 2451-9456
DOI
10.1016/j.chembiol.2025.12.010
PMC
PMC12918760
OpenAlex
W7123473986
Cited by
6
References
3
Citations by year
2026: 5
FWCI
10.33
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell chemical biology (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed Central (submittedVersion)
  • UWA Profiles and Research Repository (UWA) (submittedVersion, cc-by-nc-nd)
Funders
  • National Health and Medical Research Council (APP1147496)
  • Australian Research Council (DP220103667, DE240101210, LE230100156, FT180100204, LE120100092, LE140100096)
  • Cancer Research UK (CGCATF-2021/100012 + CGCATF-2021/100021)
  • National Institutes of Health National Cancer Institute (R01 CA238249, OT2CA278688 + OT2CA278692)
  • National Institutes of Health
  • Australian Cancer Research Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
Keywords
NONO, RNA-binding proteins, covalent inhibitors, chloroacetamide, crystal structure, breast cancer, estrogen receptor, transcriptional remodeling, tumor cell growth inhibition

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