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The splicing factor SC35 has an active role in transcriptional elongation

Nature Structural & Molecular Biology volume 15, pages 819–826 (2008)Cite this article

Abstract

Mounting evidence suggests that transcription and RNA processing are intimately coupled in vivo, although each process can occur independently in vitro. It is generally thought that polymerase II (Pol II) C-terminal domain (CTD) kinases are recruited near the transcription start site to overcome initial Pol II pausing events, and that stably bound kinases facilitate productive elongation and co-transcriptional RNA processing. Whereas most studies have focused on how RNA processing machineries take advantage of the transcriptional apparatus to efficiently modify nascent RNA, here we report that a well-studied splicing factor, SC35, affects transcriptional elongation in a gene-specific manner. SC35 depletion induces Pol II accumulation within the gene body and attenuated elongation, which are correlated with defective P-TEFb (a complex composed of CycT1–CDK9) recruitment and dramatically reduced CTD Ser2 phosphorylation. Recombinant SC35 is sufficient to rescue this defect in nuclear run-on experiments. These findings suggest a reciprocal functional relationship between the transcription and splicing machineries during gene expression.

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Figure 1: SR proteins are required for Pol II transcription in MEFs.
Figure 2: Induced Pol II accumulation in gene bodies in response to SC35 depletion in vivo.
Figure 3: Requirement for SC35 in transcription elongation.
Figure 4: Functional rescue of SC35 depletion–induced blockage of transcriptional elongation.
Figure 5: Dynamic and SC35-dependent recruitment of P-TEFb to the elongating Pol II complex.
Figure 6: Proposed model for the role of SR proteins in transcriptional elongation and co-transcriptional RNA splicing.

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Acknowledgements

We are grateful to Q. Zhou (Univeristy of California, Berkeley) and K. Jones (The Salk Institute) for advice, reporter plasmids and antibodies, K. Hertel (University of California, Irvine) for recombinant 9G8 and J. Hagopian (University of California, San Diego) for recombinant SF2/ASF. We thank M. Ares (University of California, Santa Cruz), B. Hamilton (University of California, San Diego), C. Glass (University of California, San Diego) and M.G. Rosenfeld (University of California, San Diego) for critical reading of the manuscript. This work was supported by a US National Institutes of Health (NIH) postdoctoral fellowship to G.C.-M. (1F32 GM077907) and NIH grants to X.-D.F. (5RO1 GM49369).

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Author notes

  1. Shengrong Lin

    Present address: Present address: Stanford Genome Technology Center, Stanford University, Palo Alto, California 94305, USA.,

  2. Shengrong Lin, Gabriela Coutinho-Mansfield and Dong Wang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cellular and Molecular Medicine, University of California, San Diego, 9500 Gilman Drive, La Jolla, 92093-0651, California, USA

    Shengrong Lin, Gabriela Coutinho-Mansfield, Dong Wang, Shatakshi Pandit & Xiang-Dong Fu

  2. Molecular Pathology Graduate Program, University of California, San Diego, 9500 Gilman Drive, La Jolla, 92093-0651, California, USA

    Shengrong Lin

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Contributions

X.-D.F. and S.L. designed the experiment; S.L. developed the modified nuclear run-on assay and performed most initial experiments; G.C.-M. repeated most experiments; D.W. performed the ChIP-DSL analysis; S.P. contributed to analysis of nascent RNA; all authors participated in writing the paper.

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Correspondence to Xiang-Dong Fu.

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Lin, S., Coutinho-Mansfield, G., Wang, D. et al. The splicing factor SC35 has an active role in transcriptional elongation. Nat Struct Mol Biol 15, 819–826 (2008). https://doi.org/10.1038/nsmb.1461

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