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PMID: 16396608 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Intronic binding sites for hnRNP A/B and hnRNP F/H proteins stimulate pre-mRNA splicing.

PLoS biology ·Vol. 4 ·No. 2 ·2006-02-00 ·Pages e21

Martinez-Contreras R, Fisette JF, Nasim FU, Madden R, Cordeau M, Chabot B

Abstract

hnRNP A/B proteins modulate the alternative splicing of several mammalian and viral pre-mRNAs, and are typically viewed as proteins that enforce the activity of splicing silencers. Here we show that intronic hnRNP A/B-binding sites (ABS) can stimulate the in vitro splicing of pre-mRNAs containing artificially enlarged introns. Stimulation of in vitro splicing could also be obtained by providing intronic ABS in trans through the use of antisense oligonucleotides containing a non-hybridizing ABS-carrying tail. ABS-tailed oligonucleotides also improved the in vivo inclusion of an alternative exon flanked by an enlarged intron. Notably, binding sites for hnRNP F/H proteins (FBS) replicate the activity of ABS by improving the splicing of an enlarged intron and by modulating 5' splice-site selection. One hypothesis formulated to explain these effects is that bound hnRNP proteins self-interact to bring in closer proximity the external pair of splice sites. Consistent with this model, positioning FBS or ABS at both ends of an intron was required to stimulate splicing of some pre-mRNAs. In addition, a computational analysis of the configuration of putative FBS and ABS located at the ends of introns supports the view that these motifs have evolved to support cooperative interactions. Our results document a positive role for the hnRNP A/B and hnRNP F/H proteins in generic splicing, and suggest that these proteins may modulate the conformation of mammalian pre-mRNAs.

MeSH Terms
Base Sequence Binding Sites Heterogeneous-Nuclear Ribonucleoprotein Group A-B/metabolism Heterogeneous-Nuclear Ribonucleoprotein Group F-H/metabolism Introns/genetics Models, Genetic Molecular Sequence Data Oligonucleotides, Antisense/genetics RNA Precursors/genetics RNA Splicing/genetics
Chemicals
Heterogeneous-Nuclear Ribonucleoprotein Group A-B Heterogeneous-Nuclear Ribonucleoprotein Group F-H Oligonucleotides, Antisense RNA Precursors
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Martinez-Contreras Rebeca
RNA/RNP Group, Département de Microbiologie et d'Infectiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Sherbrooke, Québec, Canada.
Fisette Jean-François
Nasim Faiz-ul Hassan
Madden Richard
Cordeau Mélanie
Chabot Benoit
Conflict of Interest

Competing interests. The authors have declared that no competing interests exist.

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Article Info
Journal
PLoS biology
Abbr.
PLoS Biol
ISSN
1545-7885
DOI
10.1371/journal.pbio.0040021
Published
2006-02-00
Epub
2006-00-10
Pages
e21
Language
English
Region
United States
NLM ID
101183755
PMCID
PMC1326234
Subset
IM
Corrections
CommentIn
PMID: 20076525
CommentIn
PMID: 20076527
External Links
PubMed source DOI: 10.1371/journal.pbio.0040021 PMC Full Text View journal details
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