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PMID: 8395005 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mutations in a protein tyrosine phosphatase gene (PTP2) and a protein serine/threonine phosphatase gene (PTC1) cause a synthetic growth defect in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 13 ·No. 9 ·1993-09-00 ·Pages 5408-17

Maeda T, Tsai AY, Saito H

Abstract

Two protein tyrosine phosphatase genes, PTP1 and PTP2, are known in Saccharomyces cerevisiae. However, the functions of these tyrosine phosphatases are unknown, because mutations in either or both phosphatase genes have no clear phenotypic effects. In this report, we demonstrate that although ptp2 has no obvious phenotype by itself, it has a profound effect on cell growth when combined with mutations in a novel protein phosphatase gene. Using a colony color sectoring assay, we isolated 25 mutants in which the expression of PTP1 or PTP2 is required for growth. Complementation tests of the mutants showed that they have a mutation in one of three genes. Cloning and sequence determination of one of these gene, PTC1, indicated that it encodes a homolog of the mammalian protein serine/threonine phosphatase 2C (PP2C). The amino acid sequence of the PTC1 product is approximately 35% identical to PP2C. Disruption of PTC1 indicated that the PTC1 function is nonessential. In contrast, ptc1 ptp2 double mutants showed a marked growth defect. To examine whether PTC1 encodes an active protein phosphatase, a glutathione S-transferase (GST)-PTC1 fusion gene was constructed and expressed in Escherichia coli. Purified GST-PTC1 fusion protein hydrolyzed a serine phosphorylated substrate in the presence of the divalent cation Mg2+ or Mn2+. GST-PTC1 also had weak (approximately 0.5% of its serine phosphatase activity) protein tyrosine phosphatase activity.

MeSH Terms
Amino Acid Sequence Base Sequence Chromosome Mapping Cloning, Molecular Fungal Proteins/genetics Genes, Dominant Genes, Fungal Genes, Lethal Genetic Complementation Test Genetic Linkage Molecular Sequence Data Mutagenesis, Insertional Phosphoprotein Phosphatases/genetics Protein Phosphatase 2 Protein Phosphatase 2C Protein Tyrosine Phosphatases/genetics Saccharomyces cerevisiae/enzymology,genetics,growth & development Saccharomyces cerevisiae Proteins Sequence Alignment Signal Transduction
Chemicals
Fungal Proteins Saccharomyces cerevisiae Proteins PTC1 protein, S cerevisiae Phosphoprotein Phosphatases Protein Phosphatase 2 Protein Phosphatase 2C Protein Tyrosine Phosphatases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Maeda T
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, Massachusetts.
Tsai A Y
Saito H
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
1993-09-00
Pages
5408-17
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360246
Subset
IM
Grants
NCI NIH HHS · CA-51132 · United States
Databases
GENBANK
L14593
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