Mutations in the SHR5 gene of Saccharomyces cerevisiae suppress Ras function and block membrane attachment and palmitoylation of Ras proteins

Jung, V., Chen, L., Hofmann, S. L., Wigler, M. H., Powers, S. (1995) Mutations in the SHR5 gene of Saccharomyces cerevisiae suppress Ras function and block membrane attachment and palmitoylation of Ras proteins. Mol Cell Biol, 15 (3). pp. 1333-42. ISSN 0270-7306 (Print)

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URL: https://www.ncbi.nlm.nih.gov/pubmed/7532279
DOI: 10.1128/MCB.15.3.1333

Abstract

We have identified a gene, SHR5, in a screen for extragenic suppressors of the hyperactive RAS2Val-19 mutation in the budding yeast Saccharomyces cerevisiae. SHR5 was cloned, sequenced, and found to encode a 23-kDa protein not significantly homologous to other proteins in the current data bases. Genetic evidence arguing that Shr5 operates at the level of Ras is presented. We tested whether SHR5, like previously isolated suppressors of hyperactivated RAS2, acts by affecting the membrane attachment and/or posttranslational modification of Ras proteins. We found that less Ras protein is attached to the membrane in shr5 mutants than in wild-type cells and that the Ras proteins are markedly underpalmitoylated, suggesting that Shr5 is involved in palmitoylation of Ras proteins. However, shr5null mutants exhibit normal palmitoyltransferase activity measured in vitro. Further, shr5null mutations attenuate Ras function in cells containing mutant Ras2 proteins that are not palmitoylated or farnesylated. We conclude that SHR5 encodes a protein that participates in the membrane localization of Ras but also interacts in vivo with completely unprocessed and cytosolic Ras proteins.

Item Type: Paper
Uncontrolled Keywords: Acyltransferases Amino Acid Sequence Base Sequence Cell Membrane metabolism Cloning Molecular Comparative Study DNA Primers Epitopes analysis Fungal Proteins biosynthesis genetics isolation & purification metabolism GTP-Binding Proteins metabolism Genes, Fungal Genotype Immunoblotting Molecular Sequence Data Mutagenesis Open Reading Frames Palmitic Acid Palmitic Acids metabolism Polymerase Chain Reaction Protein Processing Post-Translational Recombinant Proteins biosynthesis/isolation & purification Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Saccharomyces cerevisiae genetics growth & development metabolism Saccharomyces cerevisiae Proteins Suppression Genetic Temperature Valine ras Proteins
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > RAS
organism description > yeast > Saccharomyces
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mutations
CSHL Authors:
Communities: CSHL labs > Powers lab
CSHL labs > Wigler lab
Depositing User: CSHL Librarian
Date Deposited: 12 Apr 2012 17:00
Last Modified: 04 Nov 2016 20:05
PMCID: PMC230357
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26203

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