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PMID: 2835658 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning of the RNA8 gene of Saccharomyces cerevisiae, detection of the RNA8 protein, and demonstration that it is essential for nuclear pre-mRNA splicing.

Molecular and cellular biology ·Vol. 8 ·No. 3 ·1988-03-00 ·Pages 1067-75

Jackson SP, Lossky M, Beggs JD

Abstract

Strains of Saccharomyces cerevisiae that bear the temperature-sensitive mutation rna8-1 are defective in nuclear pre-mRNA splicing at the restrictive temperature (36 degrees C), suggesting that the RNA8 gene encodes a component of the splicing machinery. The RNA8 gene was cloned by complementation of the temperature-sensitive growth defect of an rna8-1 mutant strain. Integrative transformation and gene disruption experiments confirmed the identity of the cloned DNA and demonstrated that the RNA8 gene encodes an essential function. The RNA8 gene was shown to be represented once per S. cerevisiae haploid genome and to encode a low-abundance transcript of approximately 7.4 kilobases. By using antisera raised against beta-galactosidase-RNA8 fusion proteins, the RNA8 gene product was identified in S. cerevisiae cell extracts as a low-abundance protein of approximately 260 kilodaltons. Immunodepletion of the RNA8 protein specifically abolished the activity of S. cerevisiae in vitro splicing extracts, confirming that RNA8 plays an essential role in splicing.

MeSH Terms
Animals Cloning, Molecular DNA Restriction Enzymes Female Fungal Proteins/analysis,genetics Genes, Fungal Genetic Complementation Test Immune Sera Immunoassay Mutation Nucleic Acid Hybridization Plasmids RNA Precursors/genetics RNA Splicing RNA, Fungal/genetics Rabbits Saccharomyces cerevisiae/genetics Temperature Transcription, Genetic Transformation, Genetic
Chemicals
Fungal Proteins Immune Sera RNA Precursors RNA, Fungal DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Jackson S P
Department of Molecular Biology, University of Edinburgh, United Kingdom.
Lossky M
Beggs J D
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.8.3.1067-1075.1988
Published
1988-03-00
Pages
1067-75
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC363249
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.8.3.1067-1075.1988 PMC Full Text View journal details
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