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Donor site competition is involved in the regulation of alternative splicing of the rat β-tropomyosin pre-mRNA

Published online by Cambridge University Press:  01 February 1999

CHARLIE DEGUI CHEN
Affiliation:
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA Molecular and Cellular Biology Program, State University of New York at Stony Brook, Stony Brook, New York 11790, USA
DAVID M. HELFMAN
Affiliation:
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA
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Abstract

The rat β-tropomyosin (β-TM) gene encodes both skeletal muscle β-TM mRNA and nonmuscle TM-1 mRNA via alternative RNA splicing. This gene contains eleven exons: exons 1–5, 8, and 9 are common to both mRNAs; exons 6 and 11 are used in fibroblasts as well as in smooth muscle, whereas exons 7 and 10 are used in skeletal muscle. Previously we demonstrated that utilization of the 3′ splice site of exon 7 is blocked in nonmuscle cells. In this study, we use both in vitro and in vivo methods to investigate the regulation of the 5′ splice site of exon 7 in nonmuscle cells. The 5′ splice site of exon 7 is used efficiently in the absence of flanking sequences, but its utilization is suppressed almost completely when the upstream exon 6 and intron 6 are present. The suppression of the 5′ splice site of exon 7 does not result from the sequences at the 3′ end of intron 6 that block the use of the 3′ splice site of exon 7. However, mutating two conserved nucleotides GU at the 5′ splice site of exon 6 results in the efficient use of the 5′ splice site of exon 7. In addition, a mutation that changes the 5′ splice site of exon 7 to the consensus U1 snRNA binding site strongly stimulates the splicing of exon 7 to the downstream common exon 8. Collectively, these studies demonstrate that 5′ splice site competition is responsible, in part, for the suppression of exon 7 usage in nonmuscle cells.

Type
Research Article
Information
RNA , Volume 5 , Issue 2 , February 1999 , pp. 290 - 301
Copyright
1999 RNA Society

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Donor site competition is involved in the regulation of alternative splicing of the rat β-tropomyosin pre-mRNA
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