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PMID: 10990457 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

beta-subunits of Snf1 kinase are required for kinase function and substrate definition.

The EMBO journal ·Vol. 19 ·No. 18 ·2000-09-15 ·Pages 4936-43

Schmidt MC, McCartney RR

Abstract

The Snf1 kinase and its mammalian homolog, the AMP-activated protein kinase, are heterotrimeric enzymes composed of a catalytic alpha-subunit, a regulatory gamma-subunit and a beta-subunit that mediates heterotrimer formation. Saccharomyces cerevisiae encodes three beta-subunit genes, SIP1, SIP2 and GAL83. Earlier studies suggested that these subunits may not be required for Snf1 kinase function. We show here that complete and precise deletion of all three beta-subunit genes inactivates the Snf1 kinase. The sip1Delta sip2Delta gal83Delta strain is unable to derepress invertase, grows poorly on alternative carbon sources and fails to direct the phosphorylation of the Mig1 and Sip4 proteins in vivo. The SIP1 sip2Delta gal83Delta strain manifests a subset of Snf phenotypes (Raf(+), Gly(-)) observed in the snf1Delta 10 strain (Raf(-), Gly(-)), suggesting that individual beta-subunits direct the Snf1 kinase to a subset of its targets in vivo. Indeed, deletion of individual beta-subunit genes causes distinct differences in the induction and phosphorylation of Sip4, strongly suggesting that the beta-subunits play an important role in substrate definition.

MeSH Terms
AMP-Activated Protein Kinases Basic-Leucine Zipper Transcription Factors Blotting, Western Cell Division/genetics DNA-Binding Proteins/metabolism Epitopes/chemistry Fungal Proteins/metabolism Gene Deletion Genetic Complementation Test Glucose/pharmacology Glycoside Hydrolases/metabolism Models, Biological Mutagenesis, Site-Directed Phenotype Phosphorylation Protein Serine-Threonine Kinases/chemistry,genetics Protein Structure, Tertiary Repressor Proteins/metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins Trans-Activators/metabolism beta-Fructofuranosidase
Chemicals
Basic-Leucine Zipper Transcription Factors DNA-Binding Proteins Epitopes Fungal Proteins MIG1 protein, S cerevisiae Repressor Proteins SIP4 protein, S cerevisiae Saccharomyces cerevisiae Proteins Trans-Activators SNF1-related protein kinases Protein Serine-Threonine Kinases SIP1 protein, S cerevisiae AMP-Activated Protein Kinases Glycoside Hydrolases beta-Fructofuranosidase Glucose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schmidt M C
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA. mcs2@pitt.edu
McCartney R R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.18.4936
Email
mcs2@pitt.edu
Published
2000-09-15
Pages
4936-43
Language
English
Region
England
NLM ID
8208664
PMCID
PMC314222
Subset
IM
Grants
NIGMS NIH HHS · R01 GM046443 · United States
NIGMS NIH HHS · GM46443 · United States
External Links
PubMed source DOI: 10.1093/emboj/19.18.4936 PMC Full Text View journal details
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