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PMID: 9121458 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The Snf1 protein kinase and its activating subunit, Snf4, interact with distinct domains of the Sip1/Sip2/Gal83 component in the kinase complex.

Molecular and cellular biology ·Vol. 17 ·No. 4 ·1997-04-00 ·Pages 2099-106

Jiang R, Carlson M

Abstract

The Snf1 protein kinase plays a central role in the response to glucose starvation in the yeast Saccharomyces cerevisiae. Previously, we showed that two-hybrid interaction between Snf1 and its activating subunit, Snf4, is inhibited by high levels of glucose. These findings, together with biochemical evidence that Snf1 and Snf4 remain associated in cells grown in glucose, suggested that another protein (or proteins) anchors Snf1 and Snf4 into a complex. Here, we examine the possibility that a family of proteins, comprising Sip1, Sip2, and Gal83, serves this purpose. We first show that the fraction of cellular Snf4 protein that is complexed with Snf1 is reduced in a sip1delta sip2delta gal83delta triple mutant. We then present evidence that Sip1, Sip2, and Gal83 each interact independently with both Snf1 and Snf4 via distinct domains. A conserved internal region binds to the Snf1 regulatory domain, and the conserved C-terminal ASC domain binds to Snf4. Interactions were mapped by using the two-hybrid system and were confirmed by in vitro binding studies. These findings indicate that the Sip1/Sip2/Gal83 family anchors Snf1 and Snf4 into a complex. Finally, the interaction of the yeast Sip2 protein with a plant Snf1 homolog suggests that this function is conserved in plants.

MeSH Terms
AMP-Activated Protein Kinases Amino Acid Sequence Binding Sites/genetics Carrier Proteins Conserved Sequence Fungal Proteins/chemistry,genetics,metabolism Glucose/metabolism Models, Biological Molecular Sequence Data Mutation Plant Proteins/chemistry,genetics,metabolism Protein Conformation Protein Kinases/chemistry,genetics,metabolism Protein Serine-Threonine Kinases/chemistry,genetics,metabolism Repressor Proteins Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Trans-Activators Transcription Factors/chemistry,genetics,metabolism
Chemicals
Carrier Proteins Fungal Proteins GAL83 protein, S cerevisiae Plant Proteins Repressor Proteins SIP2 protein, S cerevisiae Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors Protein Kinases SNF1-related protein kinases Protein Serine-Threonine Kinases SIP1 protein, S cerevisiae SNF4 protein, S cerevisiae AMP-Activated Protein Kinases Glucose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Jiang R
Department of Genetics and Development, Columbia University, New York, New York 10032, USA.
Carlson M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.17.4.2099
Published
1997-04-00
Pages
2099-106
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC232057
Subset
IM
Grants
NIGMS NIH HHS · GM34095 · United States
External Links
PubMed source DOI: 10.1128/MCB.17.4.2099 PMC Full Text View journal details
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