Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro

Krainer, A. R., Maniatis, T. (October 1985) Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro. Cell, 42 (3). pp. 725-36. ISSN 0092-8674 (Print)0092-8674

Abstract

We have identified six distinct factors necessary for pre-mRNA splicing in vitro by selective inactivation and complementation studies, and by fractionation procedures. Splicing factor 1 (SF1) is sensitive to micrococcal nuclease, and appears to consist of at least U1 and U2 snRNPs, since splicing is inhibited when the 5' termini of U1 and U2 snRNAs are removed by site-directed cleavage with RNAase H. SF2 is a micrococcal nuclease-resistant factor present in the nuclear extract but absent from an S100 extract. SF3 is a factor that can be preferentially inactivated by moderate heat treatment. Two additional factors (SF4A and SF4B) were identified by fractionation of the nuclear extract using spermine-agarose and CM-sepharose chromatography. SF1, SF2, and SF4B appear to be required for cleavage of the pre-mRNA at the 5' splice site and lariat formation, whereas SF3 and SF4A are only required for cleavage at the 3' splice site and exon ligation.

Item Type:	Paper
Uncontrolled Keywords:	Base Sequence Endoribonucleases Globins/genetics HeLa Cells/cytology Humans Micrococcal Nuclease Nucleic Acid Precursors/*genetics RNA Precursors *RNA Splicing RNA, Messenger/*genetics Ribonuclease H Ribonucleoproteins/*genetics/isolation & purification Ribonucleoproteins, Small Nuclear
Subjects:	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 bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > splicing factor
CSHL Authors:	Krainer, Adrian R.
Communities:	CSHL labs > Krainer lab
Depositing User:	Matt Covey
Date:	October 1985
Date Deposited:	11 Mar 2014 20:39
Last Modified:	11 Mar 2014 20:39
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/29588

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