General splicing factors SF2 and SC35 have equivalent activities in vitro, and both affect alternative 5' and 3' splice site selection

Fu, X. D., Mayeda, A., Maniatis, T., Krainer, A. R. (December 1992) General splicing factors SF2 and SC35 have equivalent activities in vitro, and both affect alternative 5' and 3' splice site selection. Proceedings of the National Academy of Sciences of the United States of America, 89 (23). pp. 11224-8. ISSN 0027-8424 (Print)0027-8424

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Abstract

The human pre-mRNA splicing factors SF2 and SC35 have similar electrophoretic mobilities, and both of them contain an N-terminal ribonucleoprotein (RNP)-type RNA-recognition motif and a C-terminal arginine/serine-rich domain. However, the two proteins are encoded by different genes and display only 31% amino acid sequence identity. Here we report a systematic comparison of the splicing activities of recombinant SF2 and SC35. We find that either protein can reconstitute the splicing activity of S100 extracts and of SC35-immunodepleted nuclear extracts. Previous studies revealed that SF2 influences alternative 5' splice site selection in vitro, by favoring proximal over distal 5' splice sites, and that the A1 protein of heterogeneous nuclear RNP counteracts this effect. We now show that SC35 has a similar effect on competing 5' splice sites and is also antagonized by A1 protein. In addition, we report that both SF2 and SC35 also favor the proximal site in a pre-mRNA containing duplicated 3' splice sites, but this effect is not modulated by A1. We conclude that SF2 and SC35 are distinct splicing factors, but they display indistinguishable splicing activities in vitro.

Item Type:	Paper
Uncontrolled Keywords:	Animals Cattle Cell Nucleus/metabolism Humans Nuclear Proteins/*metabolism *RNA Splicing RNA, Heterogeneous Nuclear/metabolism RNA-Binding Proteins/*metabolism Recombinant Proteins/metabolism *Ribonucleoproteins Spliceosomes/*metabolism Temperature
Subjects:	bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA splicing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > splice site bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > splicing factor
CSHL Authors:	Krainer, Adrian R. Mayeda, Akila
Communities:	CSHL labs > Krainer lab
Depositing User:	Matt Covey
Date:	1 December 1992
Date Deposited:	11 Mar 2014 20:05
Last Modified:	13 Sep 2019 17:03
PMCID:	PMC50522
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/29596

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