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PMID: 11121472 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Role of SR protein modular domains in alternative splicing specificity in vivo.

Nucleic acids research ·Vol. 28 ·No. 24 ·2000-12-15 ·Pages 4822-31

van Der Houven Van Oordt W, Newton K, Screaton GR, Cáceres JF

Abstract

The SR proteins constitute a family of nuclear phosphoproteins which are required for constitutive splicing and also influence alternative splicing regulation. They have a modular structure consisting of one or two RNA recognition motifs (RRMs) and a C-terminal domain, rich in arginine and serine residues. The functional role of the different domains of SR proteins in constitutive splicing activity has been extensively studied in vitro; however, their contribution to alternative splicing specificity in vivo has not been clearly established. We sought to address how the modular domains of SR proteins contribute to alternative splicing specificity. The activity of a series of chimeric proteins consisting of domain swaps between different SR proteins showed that splice site selection is determined by the nature of the RRMs and that RRM2 of SF2/ASF has a dominant role and can confer specificity to a heterologous protein. In contrast, the identity of the RS domain is not important, as the RS domains are functionally interchangeable. The contribution of the RRMs to alternative splicing specificity in vivo suggests that sequence-specific RNA binding by SR proteins is required for this activity.

MeSH Terms
Adenovirus E1A Proteins/genetics Alternative Splicing Amino Acid Motifs Fibronectins/genetics Genes, Reporter/genetics HeLa Cells Humans Nuclear Proteins/chemistry,genetics,metabolism Phosphoproteins/chemistry,genetics,metabolism Protein Structure, Tertiary RNA Precursors/genetics,metabolism RNA, Messenger/genetics,metabolism RNA-Binding Proteins/chemistry,genetics,metabolism Recombinant Fusion Proteins/chemistry,metabolism Serine-Arginine Splicing Factors Substrate Specificity Transfection
Chemicals
Adenovirus E1A Proteins Fibronectins Nuclear Proteins Phosphoproteins RNA Precursors RNA, Messenger RNA-Binding Proteins Recombinant Fusion Proteins Serine-Arginine Splicing Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
van Der Houven Van Oordt W
MRC Human Genetics Unit, Western General Hospital, Edinburgh EH4 2XU, UK and Molecular Immunology Group, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK.
Newton K
Screaton G R
Cáceres J F
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/28.24.4822
Published
2000-12-15
Pages
4822-31
Language
English
Region
England
NLM ID
0411011
PMCID
PMC115228
Subset
IM
Grants
Wellcome Trust · United Kingdom
Medical Research Council · MC_U127584479 · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/28.24.4822 PMC Full Text View journal details
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