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SF2/ASF autoregulation involves multiple layers of post-transcriptional and translational control

Nature Structural & Molecular Biology volume 17, pages 306–312 (2010)Cite this article

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Abstract

SF2/ASF is a prototypical serine- and arginine-rich protein, with important roles in splicing and other aspects of mRNA metabolism. Splicing factor, arginine/serine-rich 1 (SFRS1), the gene encoding SF2/ASF, is a potent proto-oncogene with abnormal expression in many tumors. We found that SF2/ASF negatively autoregulates its expression to maintain homeostatic levels. We characterized six alternatively spliced SF2/ASF mRNA isoforms: the major isoform encodes full-length protein, whereas the others are either retained in the nucleus or degraded by nonsense-mediated mRNA decay. Unproductive splicing accounts for only part of the autoregulation, which occurs primarily at the translational level. The effect is specific to SF2/ASF and requires RNA recognition motif 2 (RRM2). The ultraconserved 3′ untranslated region (UTR) is necessary and sufficient for downregulation. SF2/ASF overexpression shifts the distribution of target mRNA toward monoribosomes, and translational repression is partly independent of Dicer and a 5′ cap. Thus, multiple post-transcriptional and translational mechanisms are involved in fine-tuning the expression of SF2/ASF.

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Figure 1: HeLa tet-off cells with inducible SF2/ASF overexpression.
Figure 2: Alternative splicing of SF2/ASF.
Figure 3: Expression of SF2/ASF from a genomic construct.
Figure 4: The 3′ UTR is necessary and sufficient for SF2/ASF autoregulation.
Figure 5: SF2/ASF reduces the polysome association of its own mRNA.
Figure 6: IRES-dependent translation assay.

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Acknowledgements

We thank B. Vogelstein (Johns Hopkins University) for generously providing the DicerEx5/Ex5 RKO cell line, G. Hannon (Cold Spring Harbor Laboratory) for sharing the Dicer−/− and Dicer+/− ES cells, V. Racaniello (Columbia University) for the gift of HCV and CrPV IRES plasmids, C. Xue for statistical analysis, Y. Yu for advice on polysome gradients and J. Zhu for helpful discussions and communicating unpublished results. This work was supported by US National Cancer Institute grant CA13106.

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Authors and Affiliations

  1. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA

    Shuying Sun, Rahul Sinha & Adrian R Krainer

  2. Graduate Program in Molecular and Cellular Biology, Stony Brook University, Stony Brook, New York, USA

    Shuying Sun & Rahul Sinha

  3. Merck Research Laboratories, Merck & Co., Inc., Rahway, New Jersey, USA

    Zuo Zhang

  4. Department of Biochemistry and Molecular Biology, The Hebrew University Medical School, Ein Karem, Jerusalem, Israel

    Rotem Karni

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Contributions

S.S. and A.R.K. designed the experiments and wrote the manuscript; S.S. performed the experiments and analyzed the data; Z.Z., R.S. and R.K. provided key reagents and advice.

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Correspondence to Adrian R Krainer.

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Sun, S., Zhang, Z., Sinha, R. et al. SF2/ASF autoregulation involves multiple layers of post-transcriptional and translational control. Nat Struct Mol Biol 17, 306–312 (2010). https://doi.org/10.1038/nsmb.1750

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