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PMID: 10072385 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Interaction of the U1 snRNP with nonconserved intronic sequences affects 5' splice site selection.

Genes & development ·Vol. 13 ·No. 5 ·1999-03-01 ·Pages 569-80

Puig O, Gottschalk A, Fabrizio P, Séraphin B

Abstract

Intron definition and splice site selection occur at an early stage during assembly of the spliceosome, the complex mediating pre-mRNA splicing. Association of U1 snRNP with the pre-mRNA is required for these early steps. We report here that the yeast U1 snRNP-specific protein Nam8p is a component of the commitment complexes, the first stable complexes assembled on pre-mRNA. In vitro and in vivo, Nam8p becomes indispensable for efficient 5' splice site recognition when this process is impaired as a result of the presence of noncanonical 5' splice sites or the absence of a cap structure. Nam8p stabilizes commitment complexes in the latter conditions. Consistent with this, Nam8p interacts with the pre-mRNA downstream of the 5' splice site, in a region of nonconserved sequence. Substitutions in this region affect splicing efficiency and alternative splice site choice in a Nam8p-dependent manner. Therefore, Nam8p is involved in a novel mechanism by which a snRNP component can affect splice site choice and regulate intron removal through its interaction with a nonconserved sequence. This supports a model where early 5' splice recognition results from a network of interactions established by the splicing machinery with various regions of the pre-mRNA.

MeSH Terms
5' Untranslated Regions Alternative Splicing Binding Sites Fungal Proteins/genetics Introns RNA Caps RNA Precursors RNA Splicing RNA-Binding Proteins/genetics Ribonucleoprotein, U1 Small Nuclear/genetics Ribonucleoproteins, Small Nuclear Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins
Chemicals
5' Untranslated Regions Fungal Proteins RNA Caps RNA Precursors RNA-Binding Proteins Ribonucleoprotein, U1 Small Nuclear Ribonucleoproteins, Small Nuclear Saccharomyces cerevisiae Proteins NAM8 protein, S cerevisiae
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Puig O
Gene Expression Program, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany.
Gottschalk A
Fabrizio P
Séraphin B
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52 references, click to expand
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.13.5.569
Published
1999-03-01
Pages
569-80
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316504
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.13.5.569 PMC Full Text View journal details
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