Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains

Caceres, J. F., Krainer, A. R. (1993) Functional analysis of pre-mRNA splicing factor SF2/ASF structural domains. EMBO Journal, 12 (12). pp. 4715-4726. ISSN 0261-4189

URL: http://www.ncbi.nlm.nih.gov/pubmed/8223480

Abstract

Human pre-mRNA splicing factor SF2/ASF has an activity required for general splicing in vitro and promotes utilization of proximal alternative 5' splice sites in a concentration-dependent manner by opposing hnRNP A1. We introduced selected mutations in the N-terminal RNA recognition motif (RRM and the C-terminal Arg/Ser (RS) domain of SF2/ASF, and assayed the resulting recombinant proteins for constitutive and alternative splicing in vitro and for binding to pre-mRNA and mRNA. Mutants inactive in constitutive splicing can affect alternative splice site selection, demonstrating that these activities involve distinct molecular interactions. Specific protein - RNA contacts mediated by Phe56 and Phe58 in the RNP-1 submotif of the SF2/ASF RRM are essential for constitutive splicing, although they are not required for RRM-mediated binding to pre-mRNA. The RS domain is also required for constitutive splicing activity and both Arg and Ser residues are important. Analysis of domain deletion mutants demonstrated strong synergy between the RRM and a central degenerate RRM repeat in binding to RNA. These two domains are sufficient for alternative splicing activity in the absence of an RS domain.

Item Type: Paper
Uncontrolled Keywords: alternative splicing premessenger rna splicing rna-protein interactions rs domain sf2 asf nuclear ribonucleoprotein binding domain conserved family site selection poly(a)-binding protein alternative 5' polymerase-ii snrnp protein 70k protein factor sc35
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear ribonucleoprotein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA splicing
CSHL Authors:
Communities: CSHL labs > Krainer lab
Depositing User: Matt Covey
Date Deposited: 11 Dec 2013 14:58
Last Modified: 11 Dec 2013 14:58
PMCID: PMC413916
URI: http://repository.cshl.edu/id/eprint/28954

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