Heterogeneous Nuclear Ribonucleoprotein G Regulates Splice Site Selection by Binding to CC(A/C)-rich Regions in Pre-mRNA*
- Bettina Heinrich‡1,
- Zhaiyi Zhang‡§1,
- Oleg Raitskin¶,
- Michael Hiller∥,
- Natalya Benderska‡,
- Annette M. Hartmann‡,
- Laurent Bracco**,
- David Elliott‡‡,
- Shani Ben-Ari¶,
- Hermona Soreq¶,
- Joseph Sperling§§,
- Ruth Sperling¶ and
- Stefan Stamm‡§2
- ‡Institute for Biochemistry, University of Erlangen, Fahrstrasse 17, 91054 Erlangen, Germany, ¶The Hebrew University of Jerusalem, Jerusalem 91904, Israel, the ∥Institute of Computer Science, Bioinformatics Group, Albert-Ludwigs-University Freiburg, Georges-Koehler-Allee 106, 79110 Freiburg, Germany, **Exonhit, ExonHit Therapeutics, 65 Boulevard Massena, 75013 Paris, France, §§The Weizmann Institute of Science, Rehovot 76100, Israel, the ‡‡Institute of Human Genetics, The International Centre for Life, Central Parkway University of Newcastle upon Tyne, Newcastle upon Tyne NE1 3BZ, United Kingdom, and the §Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky 40536-0509
- 2 To whom correspondence should be addressed: B283 Biomedical Biological Sciences Research Bldg., 741 South Limestone, Lexington, KY 40536-0509. Fax: 859-323-1037; E-mail: stefan{at}stamms-lab.net.
Abstract
Almost every protein-coding gene undergoes pre-mRNA splicing, and the majority of these pre-mRNAs are alternatively spliced. Alternative exon usage is regulated by the transient formation of protein complexes on the pre-mRNA that typically contain heterogeneous nuclear ribonucleoproteins (hnRNPs). Here we characterize hnRNP G, a member of the hnRNP class of proteins. We show that hnRNP G is a nuclear protein that is expressed in different concentrations in various tissues and that interacts with other splicing regulatory proteins. hnRNP G is part of the supraspliceosome, where it regulates alternative splice site selection in a concentration-dependent manner. Its action on alternative exons can occur without a functional RNA-recognition motif by binding to other splicing regulatory proteins. The RNA-recognition motif of hnRNP G binds to a loose consensus sequence containing a CC(A/C) motif, and hnRNP G preferentially regulates alternative exons where this motif is clustered in close proximity. The X-chromosomally encoded hnRNP G regulates different RNAs than its Y-chromosomal paralogue RNA-binding motif protein, Y-linked (RBMY), suggesting that differences in alternative splicing, evoked by the sex-specific expression of hnRNP G and RBMY, could contribute to molecular sex differences in mammals.
Footnotes
-
↵3 The abbreviations used are: hnRNP, heterogeneous nuclear ribonucleoprotein; RRM, RNA-recognition motif; GFP, green fluorescent protein; EGFP, enhanced green fluorescent protein; RT, reverse transcription; IP, immunoprecipitated; co-IP, co-immunoprecipitated; SMN2, survival of motoneuron 2.
-
↵* This work was supported, in whole or in part, by National Institutes of Health Grants GM079549 (to R. S. and J. S.), P20 RR020171, and R21HD056195-01 (to S. S.). This work was also supported by the German Research Council, the Families of SMA, EURASNET, the Bundesministerium für Bildung und Forschung, and a grant from the Helen and Milton Kimmelman Center for Biomolecular Structure and Assembly at the Weizmann Institute of Science (to J. S.).
-
↵
The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–4.
-
↵1 Both authors should be considered first authors.
-
- Received February 13, 2009.
- Revision received March 11, 2009.
- The American Society for Biochemistry and Molecular Biology, Inc.
