Article | Published:

RS domain–splicing signal interactions in splicing of U12-type and U2-type introns

Nature Structural & Molecular Biology volume 14, pages 597603 (2007) | Download Citation

Abstract

Serine-arginine (SR) proteins are general metazoan splicing factors that contain an essential arginine/serine-rich (RS) domain. On typical U2-type introns, RS domains contact the branchpoint and 5′ splice site to promote base-pairing with U small nuclear RNAs (snRNAs). Here we analyze the role of SR proteins in splicing of U12-type introns and in the second step of U2-type intron splicing. We show that RS domains contact the branchpoint and 5′ splice site of a U12-type intron. On a U2-type intron, we find that the RS domain contacts the site of the U6 snRNA–5′ splice site interaction during the first step of splicing and shifts to contact the site of the U5 snRNA–exon 1 interaction during the second step. Our results reveal alternative interactions between the RS domain and 5′ splice site region that coincide with remodeling of the spliceosome between the two catalytic steps.

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Acknowledgements

We thank A. Krainer (Cold Spring Harbor Laboratory) for providing plasmids pSP64-P120, pET19b-SF2/ASF-RRM/ψRRM and pET19b-SF2/ASF-RG, G. Akusjarvi (Uppsala University) for providing plasmids pET15b-MS2 and pET15b-MS2-RBD1, and Sara Evans for editorial assistance. This work was supported in part by a Charles A. King Trust Fellowship to H.S. and a US National Institutes of Health grant to M.R.G. M.R.G. is an investigator of the Howard Hughes Medical Institute.

Author information

Affiliations

  1. Howard Hughes Medical Institute and Programs in Gene Function and Expression and Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.

    • Haihong Shen
    •  & Michael R Green

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Contributions

H.S. and M.R.G. designed the experiments, and H.S. performed all experiments. H.S. and M.R.G. interpreted the data and prepared the manuscript.

Note: Supplementary information is available on the Nature Structural & Molecular Biology website.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Michael R Green.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Splicing of the unimolecular counterpart of the bimolecular substrate is dependent upon addition of MS2-(ASF)RS.

  2. 2.

    Supplementary Fig. 2

    On a conventional pre-mRNA substrate, the U5 snRNA-exon 1 interaction occurs prior to catalysis.

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DOI

https://doi.org/10.1038/nsmb1263

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