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An intron element modulating 5' splice site selection in the hnRNP A1 pre-mRNA interacts with hnRNP A1.
Mol Cell Biol. 1997 Apr; 17(4):1776-86.MC

Abstract

The hnRNP A1 pre-mRNA is alternatively spliced to yield the A1 and A1b mRNAs, which encode proteins differing in their ability to modulate 5' splice site selection. Sequencing a genomic portion of the murine A1 gene revealed that the intron separating exon 7 and the alternative exon 7B is highly conserved between mouse and human. In vitro splicing assays indicate that a conserved element (CE1) from the central portion of the intron shifts selection toward the distal donor site when positioned in between the 5' splice sites of exon 7 and 7B. In vivo, the CE1 element promotes exon 7B skipping. A 17-nucleotide sequence within CE1 (CE1a) is sufficient to activate the distal 5' splice site. RNase T1 protection/immunoprecipitation assays indicate that hnRNP A1 binds to CE1a, which contains the sequence UAGAGU, a close match to the reported optimal A1 binding site, UAGGGU. Replacing CE1a by different oligonucleotides carrying the sequence UAGAGU or UAGGGU maintains the preference for the distal 5' splice site. In contrast, mutations in the AUGAGU sequence activate the proximal 5' splice site. In support of a direct role of the A1-CE1 interaction in 5'-splice-site selection, we observed that the amplitude of the shift correlates with the efficiency of A1 binding. Whereas addition of SR proteins abrogates the effect of CE1, the presence of CE1 does not modify U1 snRNP binding to competing 5' splice sites, as judged by oligonucleotide-targeted RNase H protection assays. Our results suggest that hnRNP A1 modulates splice site selection on its own pre-mRNA without changing the binding of U1 snRNP to competing 5' splice sites.

Authors+Show Affiliations

Département de Microbiologie, Faculté de Médecine, Université de Sherbrooke, Quebec, Canada. b.chabot@courrier.usherb.caNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

9121425

Citation

Chabot, B, et al. "An Intron Element Modulating 5' Splice Site Selection in the hnRNP A1 pre-mRNA Interacts With hnRNP A1." Molecular and Cellular Biology, vol. 17, no. 4, 1997, pp. 1776-86.
Chabot B, Blanchette M, Lapierre I, et al. An intron element modulating 5' splice site selection in the hnRNP A1 pre-mRNA interacts with hnRNP A1. Mol Cell Biol. 1997;17(4):1776-86.
Chabot, B., Blanchette, M., Lapierre, I., & La Branche, H. (1997). An intron element modulating 5' splice site selection in the hnRNP A1 pre-mRNA interacts with hnRNP A1. Molecular and Cellular Biology, 17(4), 1776-86.
Chabot B, et al. An Intron Element Modulating 5' Splice Site Selection in the hnRNP A1 pre-mRNA Interacts With hnRNP A1. Mol Cell Biol. 1997;17(4):1776-86. PubMed PMID: 9121425.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An intron element modulating 5' splice site selection in the hnRNP A1 pre-mRNA interacts with hnRNP A1. AU - Chabot,B, AU - Blanchette,M, AU - Lapierre,I, AU - La Branche,H, PY - 1997/4/1/pubmed PY - 1997/4/1/medline PY - 1997/4/1/entrez SP - 1776 EP - 86 JF - Molecular and cellular biology JO - Mol Cell Biol VL - 17 IS - 4 N2 - The hnRNP A1 pre-mRNA is alternatively spliced to yield the A1 and A1b mRNAs, which encode proteins differing in their ability to modulate 5' splice site selection. Sequencing a genomic portion of the murine A1 gene revealed that the intron separating exon 7 and the alternative exon 7B is highly conserved between mouse and human. In vitro splicing assays indicate that a conserved element (CE1) from the central portion of the intron shifts selection toward the distal donor site when positioned in between the 5' splice sites of exon 7 and 7B. In vivo, the CE1 element promotes exon 7B skipping. A 17-nucleotide sequence within CE1 (CE1a) is sufficient to activate the distal 5' splice site. RNase T1 protection/immunoprecipitation assays indicate that hnRNP A1 binds to CE1a, which contains the sequence UAGAGU, a close match to the reported optimal A1 binding site, UAGGGU. Replacing CE1a by different oligonucleotides carrying the sequence UAGAGU or UAGGGU maintains the preference for the distal 5' splice site. In contrast, mutations in the AUGAGU sequence activate the proximal 5' splice site. In support of a direct role of the A1-CE1 interaction in 5'-splice-site selection, we observed that the amplitude of the shift correlates with the efficiency of A1 binding. Whereas addition of SR proteins abrogates the effect of CE1, the presence of CE1 does not modify U1 snRNP binding to competing 5' splice sites, as judged by oligonucleotide-targeted RNase H protection assays. Our results suggest that hnRNP A1 modulates splice site selection on its own pre-mRNA without changing the binding of U1 snRNP to competing 5' splice sites. SN - 0270-7306 UR - https://www.unboundmedicine.com/medline/citation/9121425/An_intron_element_modulating_5'_splice_site_selection_in_the_hnRNP_A1_pre_mRNA_interacts_with_hnRNP_A1_ L2 - http://mcb.asm.org/cgi/pmidlookup?view=long&pmid=9121425 DB - PRIME DP - Unbound Medicine ER -