Branch point selection in alternative splicing of tropomyosin pre-mRNAs

Helfman, D. M., Ricci, W. M. (July 1989) Branch point selection in alternative splicing of tropomyosin pre-mRNAs. Nucleic Acids Res, 17 (14). pp. 5633-50. ISSN 0305-1048

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URL: http://www.ncbi.nlm.nih.gov/pubmed/2762151
DOI: 10.1093/nar/17.14.5633

Abstract

The rat tropomyosin 1 gene gives rise to two mRNAs encoding rat fibroblast TM-1 and skeletal muscle beta-tropomyosin via an alternative splicing mechanism. The gene is comprised of 11 exons. Exons 1 through 5 and exons 8 and 9 are common to all mRNAs expressed from this gene. Exons 6 and 11 are used in fibroblasts as well as smooth muscle whereas exons 7 and 10 are used exclusively in skeletal muscle. In the present studies we have focused on the mutually exclusive internal alternative splice choice involving exon 6 (fibroblast-type splice) and exon 7 (skeletal muscle-type splice). To study the mechanism and regulation of alternative splice site selection we have characterized the branch points used in processing of the tropomyosin pre-mRNAs in vitro using nuclear extracts obtained from HeLa cells. Splicing of exon 5 to exon 6 (fibroblast-type splice) involves the use of three branch points located 25, 29, and 36 nucleotides upstream of the 3' splice site of exon 6. Splicing of exon 6 (fibroblast-type splice) or exon 7 (skeletal muscle type-splice) to exon 8 involves the use of the same branch point located 24 nucleotides upstream of this shared 3' splice site. In contrast, the splicing of exon 5 to exon 7 (skeletal muscle-type splice) involves the use of three branch sites located 144, 147 and 153 nucleotides, upstream of the 3' splice site of exon 7. In addition, the pyrimidine content of the region between these unusual branch points and the 3' splice site of exon 7 was found to be greater than 80%. These studies raise the possibility that the use of branch points located a long distance from a 3' splice site may be an essential feature of some alternatively spliced exons. The possible significance of these unusual branch points as well as a role for the polypyrimidine stretch in intron 6 in splice site selection are discussed.

Item Type: Paper
Uncontrolled Keywords: Animals Base Sequence Exons Fibroblasts/metabolism *Genes, Structural Introns Molecular Sequence Data Muscles/metabolism Plasmids RNA Precursors/*genetics *RNA Splicing Rats Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Templates, Genetic Transcription, Genetic Tropomyosin/*genetics
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Alternative Splicing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
CSHL Authors:
Communities: CSHL labs > Helfman lab
Depositing User: Gail Sherman
Date: 25 July 1989
Date Deposited: 27 Jul 2017 19:34
Last Modified: 08 Nov 2017 16:58
PMCID: PMC318185
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34860

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