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A highly stable duplex structure sequesters the 5' splice site region of hnRNP A1 alternative exon 7B.
RNA. 1997 Apr; 3(4):405-19.RNA

Abstract

Exon 7B in the hnRNP A1 pre-mRNA is alternatively spliced to yield A1 and A1(B), two proteins that differ in their ability to modulate 5' splice site selection. Sequencing the murine intron downstream of exon 7B revealed the existence of several regions of similarity to the corresponding human intron. In vitro splicing assays indicate that an 84-nt region (CE6IO) decreases splicing to the proximal 5' splice site in a pre-mRNA carrying the 5' splice sites of exon 7 and 7B. In vivo, the CE6IO element promotes exon 7B skipping in pre-mRNAs expressed from a mini-gene containing the hnRNP A1 alternative splicing unit. Using oligonucleotide-targeted RNase H cleavage assays, we provide support for the existence of highly stable base pairing interactions between CE6IO and the 5' splice site region of exon 7B. Duplex formation occurs in naked pre-mRNA, resists incubation in splicing extracts, and is associated with a reduction in the assembly of U1 snRNP-dependent complexes to the 5' splice site of exon 7B. Our results demonstrate that pre-mRNA secondary structure plays an important role in promoting exon 7B skipping in the A1 pre-mRNA.

Authors+Show Affiliations

Départment de Microbiologie et Infectiologie, Faculté de Médicine, Université de Sherbrooke, Québec, Canada.No affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9085847

Citation

Blanchette, M, and B Chabot. "A Highly Stable Duplex Structure Sequesters the 5' Splice Site Region of hnRNP A1 Alternative Exon 7B." RNA (New York, N.Y.), vol. 3, no. 4, 1997, pp. 405-19.
Blanchette M, Chabot B. A highly stable duplex structure sequesters the 5' splice site region of hnRNP A1 alternative exon 7B. RNA. 1997;3(4):405-19.
Blanchette, M., & Chabot, B. (1997). A highly stable duplex structure sequesters the 5' splice site region of hnRNP A1 alternative exon 7B. RNA (New York, N.Y.), 3(4), 405-19.
Blanchette M, Chabot B. A Highly Stable Duplex Structure Sequesters the 5' Splice Site Region of hnRNP A1 Alternative Exon 7B. RNA. 1997;3(4):405-19. PubMed PMID: 9085847.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A highly stable duplex structure sequesters the 5' splice site region of hnRNP A1 alternative exon 7B. AU - Blanchette,M, AU - Chabot,B, PY - 1997/4/1/pubmed PY - 1997/4/1/medline PY - 1997/4/1/entrez SP - 405 EP - 19 JF - RNA (New York, N.Y.) JO - RNA VL - 3 IS - 4 N2 - Exon 7B in the hnRNP A1 pre-mRNA is alternatively spliced to yield A1 and A1(B), two proteins that differ in their ability to modulate 5' splice site selection. Sequencing the murine intron downstream of exon 7B revealed the existence of several regions of similarity to the corresponding human intron. In vitro splicing assays indicate that an 84-nt region (CE6IO) decreases splicing to the proximal 5' splice site in a pre-mRNA carrying the 5' splice sites of exon 7 and 7B. In vivo, the CE6IO element promotes exon 7B skipping in pre-mRNAs expressed from a mini-gene containing the hnRNP A1 alternative splicing unit. Using oligonucleotide-targeted RNase H cleavage assays, we provide support for the existence of highly stable base pairing interactions between CE6IO and the 5' splice site region of exon 7B. Duplex formation occurs in naked pre-mRNA, resists incubation in splicing extracts, and is associated with a reduction in the assembly of U1 snRNP-dependent complexes to the 5' splice site of exon 7B. Our results demonstrate that pre-mRNA secondary structure plays an important role in promoting exon 7B skipping in the A1 pre-mRNA. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/9085847/A_highly_stable_duplex_structure_sequesters_the_5'_splice_site_region_of_hnRNP_A1_alternative_exon_7B_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=9085847 DB - PRIME DP - Unbound Medicine ER -