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Despite a number of efforts to target RBPs with small molecules, it is still difficult to develop RBP inhibitors, asking for a deeper understanding of how to chemically perturb RNA-binding activity. In this study, we found that the thiopurine drugs (6-mercaptopurine and 6-thioguanine) effectively disrupt CELF1\u2013RNA interaction. The disrupting activity relies on the formation of disulfide bonds between the thiopurine drugs and CELF1. Mutating the cysteine residue proximal to the RNA recognition motifs (RRMs), or adding reducing agents, abolishes the disrupting activity. Furthermore, the 1,2,4-triazole-3-thione, a thiopurine analogue, was identified with 20-fold higher disrupting activity. Based on this analogue, we found that compound 9 disrupts CELF1\u2013RNA interaction in living cells and ameliorates CELF1-mediated myogenesis deficiency. In summary, we identified a thiol-mediated binding mechanism for thiopurine drugs and their derivatives to perturb protein-RNA interaction, which provides novel insight for developing RBP inhibitors. Additionally, this work may benefit the pharmacological and toxicity research of thiopurine drugs.<\/jats:p>","DOI":"10.1093\/nar\/gkae788","type":"journal-article","created":{"date-parts":[[2024,9,13]],"date-time":"2024-09-13T19:00:01Z","timestamp":1726254001000},"page":"10810-10822","source":"Crossref","is-referenced-by-count":3,"title":["Identification of an RNA-binding perturbing characteristic for thiopurine drugs and their derivatives to disrupt CELF1\u2013RNA interaction"],"prefix":"10.1093","volume":"52","author":[{"given":"Yang","family":"Tan","sequence":"first","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Drum Tower Hospital Affiliated to Medical School, School of Life Sciences, Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhibo","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Drum Tower Hospital Affiliated to Medical School, School of Life Sciences, Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingfang","family":"Han","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Drum Tower Hospital Affiliated to Medical School, School of Life Sciences, Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peipei","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Hematology, Drum Tower Hospital Affiliated to Medical School, Nanjing University , Nanjing 210008 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6116-635X","authenticated-orcid":false,"given":"Xiaopeng","family":"Shen","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Anhui Normal University , Wuhu ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yajun","family":"Jiang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Chemistry and Biomedicine Innovation Center (ChemBIC), Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiang","family":"Xu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Drum Tower Hospital Affiliated to Medical School, School of Life Sciences, Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0834-4601","authenticated-orcid":false,"given":"Xingxin","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Pharmaceutical Biotechnology, Drum Tower Hospital Affiliated to Medical School, School of Life Sciences, Nanjing University , Nanjing 210023 ,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,9,12]]},"reference":[{"key":"2024101409565991100_B1","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1038\/s41576-020-00302-y","article-title":"RNA-binding proteins in human genetic disease","volume":"22","author":"Gebauer","year":"2020","journal-title":"Nat. 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