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Branch point selection in alternative splicing of tropomyosin pre-mRNAs.
Nucleic Acids Res. 1989 Jul 25; 17(14):5633-50.NA

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.

Authors+Show Affiliations

Cold Spring Harbor Laboratory, NY 11724.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

2762151

Citation

Helfman, D M., and W M. Ricci. "Branch Point Selection in Alternative Splicing of Tropomyosin Pre-mRNAs." Nucleic Acids Research, vol. 17, no. 14, 1989, pp. 5633-50.
Helfman DM, Ricci WM. Branch point selection in alternative splicing of tropomyosin pre-mRNAs. Nucleic Acids Res. 1989;17(14):5633-50.
Helfman, D. M., & Ricci, W. M. (1989). Branch point selection in alternative splicing of tropomyosin pre-mRNAs. Nucleic Acids Research, 17(14), 5633-50.
Helfman DM, Ricci WM. Branch Point Selection in Alternative Splicing of Tropomyosin Pre-mRNAs. Nucleic Acids Res. 1989 Jul 25;17(14):5633-50. PubMed PMID: 2762151.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Branch point selection in alternative splicing of tropomyosin pre-mRNAs. AU - Helfman,D M, AU - Ricci,W M, PY - 1989/7/25/pubmed PY - 1989/7/25/medline PY - 1989/7/25/entrez SP - 5633 EP - 50 JF - Nucleic acids research JO - Nucleic Acids Res VL - 17 IS - 14 N2 - 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. SN - 0305-1048 UR - https://www.unboundmedicine.com/medline/citation/2762151/Branch_point_selection_in_alternative_splicing_of_tropomyosin_pre_mRNAs_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/17.14.5633 DB - PRIME DP - Unbound Medicine ER -