Abstract
Pre-mRNA splicing is functionally coupled to transcription, and genotoxic stresses can enhance alternative exon inclusion by affecting elongating RNA polymerase II. We report here that various genotoxic stress inducers, including camptothecin (CPT), inhibit the interaction between Ewing's sarcoma proto-oncoprotein (EWS), an RNA polymerase II–associated factor, and YB-1, a spliceosome-associated factor. This results in the cotranscriptional skipping of several exons of the MDM2 gene, which encodes the main p53 ubiquitin ligase. This reversible exon skipping participates in the regulation of MDM2 expression that may contribute to the accumulation of p53 during stress exposure and its rapid shut-off when stress is removed. Finally, a splicing-sensitive microarray identified numerous exons that are skipped in response to CPT and EWS–YB-1 depletion. These data demonstrate genotoxic stress-induced alteration of the communication between the transcriptional and splicing machineries, which results in widespread exon skipping and plays a central role in the genotoxic stress response.
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Acknowledgements
We are grateful to Angela Krämer (University of Geneva) and Robin Fahraeus (Institut National de la Santé et de la Recherche Médicale U940) for providing antibodies. This work was supported by the EC (NoE EURASNET), Association pour la recherche sur le cancer, Ligue nationale contre le cancer, Institut national du cancer and Agence Nationale de la Recherche. G.S. was supported by La Chancellerie des Universités de Paris; M.D. by l'Institut National de la Santé et de la Recherche Médicale; J.B. by the French Ministry of Education. M.-C.D.C. was supported by a fellowship from the Brittany Region; G.D. by Vaincre la Mucoviscidose; L.C. by Brest University, the IFR 148-ScInBioS.
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M.D., G.S., M.-C.D.C., L.C. and D.A. designed the experiments; M.D., L.C. and D.A. supervised the project; M.D., G.S., M.-C.D.C., J.B., E.D., L.G., G.D. and C.L.-C. conducted the experiments; M.D., G.S., M.-C.D.C., C.L.-C., M.-C.D.C., L.C. and D.A. performed the data analysis; M.D., G.S., M.-C.D.C., L.C. and D.A. wrote the manuscript.
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Dutertre, M., Sanchez, G., De Cian, MC. et al. Cotranscriptional exon skipping in the genotoxic stress response. Nat Struct Mol Biol 17, 1358–1366 (2010). https://doi.org/10.1038/nsmb.1912
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DOI: https://doi.org/10.1038/nsmb.1912
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