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PMID: 3517868 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Fractionation and characterization of a yeast mRNA splicing extract.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 8 ·1986-04-00 ·Pages 2387-91

Cheng SC, Abelson J

Abstract

We have fractionated a yeast whole cell extract that can accurately splice synthetic actin and CYH2 pre-mRNAs. Three fractions, designated I, II, and III, have been separated by use of ammonium sulfate fractionation and chromatography on heparin agarose. Each fraction alone has no splicing activity. Fractions I and II allow the first step of the splicing reaction to proceed, giving rise to the splicing intermediates, free exon 1, and intron-exon 2. Addition of fraction III completes the reaction. Micrococcal nuclease treatment of the whole cell extract or of either fraction I or II abolished splicing activity, indicating that fractions I and II have RNA moieties that are required in the splicing reaction. The nature of the RNAs was examined using antibodies directed against the trimethylated cap structure unique to small nuclear RNAs. Preincubation of the whole cell extract with protein A-Sepharose coupled to trimethylated cap antibody abolished splicing activity. This indicates that at least one essential RNA component contains a trimethyl cap. Thus, in yeast as in mammalian systems, small nuclear RNAs are involved in mRNA splicing.

MeSH Terms
Cell Fractionation RNA Caps/analysis,immunology RNA Processing, Post-Transcriptional RNA Splicing RNA, Small Nuclear/genetics Saccharomyces cerevisiae/genetics
Chemicals
RNA Caps RNA, Small Nuclear
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Cheng S C
Abelson J
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30 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.8.2387
Published
1986-04-00
Pages
2387-91
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323302
Subset
IM
Grants
NIGMS NIH HHS · GM 30356 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.8.2387 PMC Full Text View journal details
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