Multiple functional domains of the yeast l,3-beta-glucan synthase subunit Fks1p revealed by quantitative phenotypic analysis of temperature-sensitive mutants

Hiroki Okada; Mitsuhiro Abe; Masayo Asakawa-Minemura; Aiko Hirata; Hiroshi Qadota; Kana Morishita; Shinsuke Ohnuki; Satoru Nogami; Yoshikazu Ohya

doi:10.1534/genetics.109.109892

Multiple functional domains of the yeast l,3-beta-glucan synthase subunit Fks1p revealed by quantitative phenotypic analysis of temperature-sensitive mutants

Genetics. 2010 Apr;184(4):1013-24. doi: 10.1534/genetics.109.109892. Epub 2010 Feb 1.

Authors

Hiroki Okada¹, Mitsuhiro Abe, Masayo Asakawa-Minemura, Aiko Hirata, Hiroshi Qadota, Kana Morishita, Shinsuke Ohnuki, Satoru Nogami, Yoshikazu Ohya

Affiliation

¹ Department of Biological Sciences, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, Japan.

Abstract

The main filamentous structural component of the cell wall of the yeast Saccharomyces cerevisiae is 1,3-beta-glucan, which is synthesized by a plasma membrane-localized enzyme called 1,3-beta-glucan synthase (GS). Here we analyzed the quantitative cell morphology and biochemical properties of 10 different temperature-sensitive mutants of FKS1, a putative catalytic subunit of GS. To untangle their pleiotropic phenotypes, the mutants were classified into three functional groups. In the first group, mutants fail to synthesize 1,3-beta-glucan at the proper subcellular location, although GS activity is normal in vitro. In the second group, mutants have normal 1,3-beta-glucan content but are defective in polarized growth and endocytosis. In the third group, mutations in the putative catalytic domain of Fks1p result in a loss of the catalytic activity of GS. The differences among the three groups suggest that Fks1p consists of multiple domains that are required for cell wall construction and cellular morphogenesis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Catalytic Domain*
Cell Wall / metabolism
Cluster Analysis
Echinocandins / chemistry
Echinocandins / classification
Echinocandins / genetics*
Echinocandins / metabolism
Endocytosis / genetics
Glucosyltransferases / chemistry*
Glucosyltransferases / classification
Glucosyltransferases / genetics*
Glucosyltransferases / metabolism
Membrane Proteins / chemistry
Membrane Proteins / classification
Membrane Proteins / genetics*
Membrane Proteins / metabolism
Mutant Proteins / chemistry
Mutant Proteins / classification
Mutant Proteins / genetics
Mutant Proteins / metabolism
Mutation*
Phenotype*
Saccharomyces cerevisiae / cytology
Saccharomyces cerevisiae / enzymology*
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism
Saccharomyces cerevisiae Proteins / chemistry
Saccharomyces cerevisiae Proteins / classification
Saccharomyces cerevisiae Proteins / genetics*
Saccharomyces cerevisiae Proteins / metabolism
Temperature*

Substances

Echinocandins
Membrane Proteins
Mutant Proteins
Saccharomyces cerevisiae Proteins
Glucosyltransferases
1,3-beta-glucan synthase
FKS1 protein, S cerevisiae
GSC2 protein, S cerevisiae