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Dephosphorylated SRp38 acts as a splicing repressor in response to heat shock

Nature volume 427, pages 553–558 (2004)Cite this article

Abstract

The cellular response to stresses such as heat shock involves changes in gene expression1. It is well known that the splicing of messenger RNA precursors is generally repressed on heat shock2,3, but the factors responsible have not been identified4,5,6,7,8. SRp38 is an SR protein splicing factor9,10 that functions as a general repressor of splicing. It is activated by dephosphorylation and required for splicing repression in M-phase cells11. Here we show that SRp38 is also dephosphorylated on heat shock and that this dephosphorylation correlates with splicing inhibition. Notably, depletion of SRp38 from heat-shocked cell extracts derepresses splicing, and adding back dephosphorylated SRp38 specifically restores inhibition. We further show that dephosphorylated SRp38 interacts with a U1 small nuclear ribonucleoprotein particle (snRNP) protein, and that this interaction interferes with 5′-splice-site recognition by the U1 snRNP. Finally, SRp38-deficient DT40 cells show an altered cell-cycle profile consistent with a mitotic defect; they are also temperature sensitive and defective in recovery after heat shock. SRp38 thus plays a crucial role in cell survival under stress conditions by inhibiting the splicing machinery.

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Figure 1: SRp38 is dephosphorylated and splicing is inhibited in response to heat shock.
Figure 2: dSRp38 is necessary for heat-shock splicing repression.
Figure 3: dSRp38 interacts with U1 snRNP and inhibits 5′-splice-site recognition.
Figure 4: Characteristics of SRp38-deficient DT40 cells.

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Acknowledgements

We thank Z. Chen, T. Kashima, S. Bush, Y. Shi and X. Li for plasmids and discussion; A. Norris and A. Doty for technical assistance; I. Boluk for help preparing the manuscript; R. Luhrmann and B. Kastner for purified U1 snRNP; and W. van Venrooij for antibodies against U1-70K. This work was supported by a grant from the NIH.

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  1. Chanseok Shin and Ying Feng: These authors contributed equally to this work

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  1. Department of Biological Sciences, Columbia University, New York, New York, 10027, USA

    Chanseok Shin, Ying Feng & James L. Manley

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  1. Chanseok Shin
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Correspondence to James L. Manley.

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Shin, C., Feng, Y. & Manley, J. Dephosphorylated SRp38 acts as a splicing repressor in response to heat shock. Nature 427, 553–558 (2004). https://doi.org/10.1038/nature02288

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