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Mutation of PTB binding sites causes misregulation of alternative 3' splice site selection in vivo.
RNA. 1997 Jul; 3(7):764-78.RNA

Abstract

Alternative splicing of pre-mRNA is a commonly used mechanism to regulate gene expression in higher eukaryotes. However, with the exception of regulated cascades in Drosophila, the cis-acting elements and the trans-acting factors that control tissue- and/or developmentally regulated splicing remain largely unidentified. Cis-acting elements that control smooth muscle-specific repression of exon 3 of alpha-tropomyosin (alpha-TM) have been identified recently and consist of two regions that flank this exon. Deletion of either element causes misregulated splicing of alpha-TM in transfected smooth muscle cells. In experiments designed to characterize essential sequences within each element and the factors that interact with these sequences, we have identified two overlapping sequences within the downstream regulatory element (DRE) that are identical to binding sites for polypyrimidine tract binding protein (PTB) that were identified using iterative selection techniques. Mutation of these sites caused aberrant splicing regulation in transfected smooth muscle cells. In addition, sequences identical to high-affinity PTB binding sites were also detected upstream of exon 3 and mutation of these sites also resulted in misregulation of splicing in vivo, suggesting that PTB binding to specific sequences flanking exon 3 is responsible, in part, for the repression of exon 3. Consistent with this hypothesis, UV crosslinking and equilibrium binding assays confirm that the same mutations that cause misregulated splicing also disrupt PTB binding to RNA.

Authors+Show Affiliations

Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.No affiliation info availableNo affiliation info availableNo 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

9214659

Citation

Pérez, I, et al. "Mutation of PTB Binding Sites Causes Misregulation of Alternative 3' Splice Site Selection in Vivo." RNA (New York, N.Y.), vol. 3, no. 7, 1997, pp. 764-78.
Pérez I, Lin CH, McAfee JG, et al. Mutation of PTB binding sites causes misregulation of alternative 3' splice site selection in vivo. RNA. 1997;3(7):764-78.
Pérez, I., Lin, C. H., McAfee, J. G., & Patton, J. G. (1997). Mutation of PTB binding sites causes misregulation of alternative 3' splice site selection in vivo. RNA (New York, N.Y.), 3(7), 764-78.
Pérez I, et al. Mutation of PTB Binding Sites Causes Misregulation of Alternative 3' Splice Site Selection in Vivo. RNA. 1997;3(7):764-78. PubMed PMID: 9214659.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mutation of PTB binding sites causes misregulation of alternative 3' splice site selection in vivo. AU - Pérez,I, AU - Lin,C H, AU - McAfee,J G, AU - Patton,J G, PY - 1997/7/1/pubmed PY - 1997/7/1/medline PY - 1997/7/1/entrez SP - 764 EP - 78 JF - RNA (New York, N.Y.) JO - RNA VL - 3 IS - 7 N2 - Alternative splicing of pre-mRNA is a commonly used mechanism to regulate gene expression in higher eukaryotes. However, with the exception of regulated cascades in Drosophila, the cis-acting elements and the trans-acting factors that control tissue- and/or developmentally regulated splicing remain largely unidentified. Cis-acting elements that control smooth muscle-specific repression of exon 3 of alpha-tropomyosin (alpha-TM) have been identified recently and consist of two regions that flank this exon. Deletion of either element causes misregulated splicing of alpha-TM in transfected smooth muscle cells. In experiments designed to characterize essential sequences within each element and the factors that interact with these sequences, we have identified two overlapping sequences within the downstream regulatory element (DRE) that are identical to binding sites for polypyrimidine tract binding protein (PTB) that were identified using iterative selection techniques. Mutation of these sites caused aberrant splicing regulation in transfected smooth muscle cells. In addition, sequences identical to high-affinity PTB binding sites were also detected upstream of exon 3 and mutation of these sites also resulted in misregulation of splicing in vivo, suggesting that PTB binding to specific sequences flanking exon 3 is responsible, in part, for the repression of exon 3. Consistent with this hypothesis, UV crosslinking and equilibrium binding assays confirm that the same mutations that cause misregulated splicing also disrupt PTB binding to RNA. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/9214659/Mutation_of_PTB_binding_sites_causes_misregulation_of_alternative_3'_splice_site_selection_in_vivo_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=9214659 DB - PRIME DP - Unbound Medicine ER -