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PMID: 8289829 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The mutant type 1 protein phosphatase encoded by glc7-1 from Saccharomyces cerevisiae fails to interact productively with the GAC1-encoded regulatory subunit.

Molecular and cellular biology ·Vol. 14 ·No. 2 ·1994-02-00 ·Pages 896-905

Stuart JS, Frederick DL, Varner CM, Tatchell K

Abstract

Loss-of-function gac1 mutants of Saccharomyces cerevisiae fail to accumulate normal levels of glycogen because of low glycogen synthase activity. Increased dosage of GAC1 results in increased activity of glycogen synthase and a corresponding hyperaccumulation of glycogen. The glycogen accumulation phenotype of gac1 is similar to that of glc7-1, a type 1 protein phosphatase mutant. We have partially characterized the GAC1 gene product (Gac1p) and show that levels of Gac1p increase during growth with the same kinetics as glycogen accumulation. Gac1p is phosphorylated in vivo and is hyperphosphorylated in a glc7-1 mutant. Gac1p and the type 1 protein phosphatase directly interact in vitro, as assayed by coimmunoprecipitation, and in vivo, as determined by the dihybrid assay described elsewhere (S. Fields and O.-k. Song, Nature [London] 340:245-246, 1989). The interaction between Gac1p and the glc7-1-encoded form of the type 1 protein phosphatase is defective, as assayed by either immunoprecipitation or the dihybrid assay. Increased dosage of GAC1 partially suppresses the glycogen defect of glc7-1. Collectively, our data support the hypotheses that GAC1 encodes a regulatory subunit of type 1 protein phosphatase and that the glycogen accumulation defect of glc7-1 is due at least in part to the inability of the mutant phosphatase to interact with its regulatory subunit.

Related Genes
MeSH Terms
Base Sequence Crosses, Genetic DNA Primers Fungal Proteins/biosynthesis,metabolism Gene Expression Isoenzymes/biosynthesis,genetics,metabolism Kinetics Molecular Sequence Data Mutation Phosphoprotein Phosphatases/biosynthesis,genetics,metabolism Plasmids Polymerase Chain Reaction Protein Phosphatase 1 Recombinant Fusion Proteins/biosynthesis,metabolism Saccharomyces cerevisiae/enzymology,genetics Saccharomyces cerevisiae Proteins beta-Galactosidase/biosynthesis,metabolism
Chemicals
DNA Primers Fungal Proteins GAC1 protein, S cerevisiae Isoenzymes Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Phosphoprotein Phosphatases Protein Phosphatase 1 beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Stuart J S
Department of Microbiology, North Carolina State University, Raleigh 27695-7615.
Frederick D L
Varner C M
Tatchell K
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
1994-02-00
Pages
896-905
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358444
Subset
IM
Grants
NIGMS NIH HHS · GM47789 · United States
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