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Molecular and Cellular Biology, August 2004, p. 6811-6823, Vol. 24, No. 15
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.15.6811-6823.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

SMU-2 and SMU-1, Caenorhabditis elegans Homologs of Mammalian Spliceosome-Associated Proteins RED and fSAP57, Work Together To Affect Splice Site Choice

Angela K. Spartz, Robert K. Herman, and Jocelyn E. Shaw*

Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455

Received 16 October 2003/ Returned for modification 13 January 2004/ Accepted 20 April 2004

Mutations in the Caenorhabditis elegans gene smu-2 suppress mec-8 and unc-52 mutations. It has been proposed that MEC-8 regulates the alternative splicing of unc-52 transcripts, which encode the core protein of perlecan, a basement membrane proteoglycan. We show that mutation in smu-2 leads to enhanced accumulation of transcripts that skip exon 17, but not exon 18, of unc-52, which explains our finding that smu-2 mutations suppress the uncoordination conferred by nonsense mutations in exon 17, but not in exon 18, of unc-52. We conclude that smu-2 encodes a ubiquitously expressed nuclear protein that is 40% identical to the human RED protein, a component of purified spliceosomes. The effects of smu-2 mutation on both unc-52 pre-mRNA splicing and the suppression of mec-8 and unc-52 mutant phenotypes are indistinguishable from the effects of mutation in smu-1, a gene that encodes a protein that is 62% identical to human spliceosome-associated protein fSAP57. We provide evidence that SMU-2 protects SMU-1 from degradation in vivo. In vitro and in vivo coimmunoprecipitation experiments indicate that SMU-2 and SMU-1 bind to each other. We propose that SMU-2 and SMU-1 function together to regulate splice site choice in the pre-mRNAs of unc-52 and other genes.

* Corresponding author. Mailing address: Department of Genetics, Cell Biology and Development, University of Minnesota, 6-160 Jackson Hall, 321 Washington Ave. SE, Minneapolis, MN 55455. Phone: (612) 625-1912. Fax: (612) 624-8118. E-mail: jocelyn{at}biosci.cbs.umn.edu.

Molecular and Cellular Biology, August 2004, p. 6811-6823, Vol. 24, No. 15
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.15.6811-6823.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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