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Open AccessJournal ArticleDOI

Chitin synthase I and chitin synthase II are not required for chitin synthesis in vivo in Saccharomyces cerevisiae.

TLDR
The double disruption mutant of Saccharomyces cerevisiae has no detectable chitin deficiency in vivo, as judged by quantitative assay and by staining cells with Calcofluor, and assay of membrane preparations from thedouble disruption mutant indicates the presence of chit in synthetic activity.
Abstract
In Saccharomyces cerevisiae, the polysaccharide chitin forms the primary division septum between mother cell and bud. Two related enzymes, chitin synthase I and chitin synthase II (UDP-acetamido-2-deoxy-D-glucose:chitin 4-beta-acetamidodeoxyglucosyltransferase, EC 2.4.1.16), have been identified and their structural genes, CHS1 and CHS2, respectively, have been cloned and sequenced. Gene disruption experiments led to the conclusion that CHS2 is essential for cell division [Silverman, S.J., Sburlati, A., Slater, M.L. & Cabib, E. (1988) Proc. Natl. Acad. Sci. USA 85, 4735-4739], whereas CHS1 is not. We repeated the disruption of CHS2 and determined that it is not essential for vegetative growth. The viability of chs1::HIS3 chs2::TRP1 spores is influenced by strain background and germination conditions. The double disruption mutant has no detectable chitin deficiency in vivo, as judged by quantitative assay and by staining cells with Calcofluor. Assay of membrane preparations from the double disruption mutant indicates the presence of chitin synthetic activity. Unlike the CHS gene products, this third activity is not stimulated by trypsin. Characterization of the double disruption mutant revealed abnormalities in morphology and nuclear migration.

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Journal ArticleDOI

Architecture of the yeast cell wall. Beta(1-->6)-glucan interconnects mannoprotein, beta(1-->)3-glucan, and chitin.

TL;DR: It is concluded that the latter polysaccharide has a central role in the organization of the yeast cell wall and the possible mechanism of synthesis and physiological significance of the cross-links is discussed.
Journal ArticleDOI

Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae.

TL;DR: An overall view of the current understanding of cell wall dynamics and of the complex network that controls polarized growth at particular stages of the budding yeast cell cycle and life cycle is offered.
Journal ArticleDOI

The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle.

TL;DR: Genetic evidence indicated that a mutant lacking all three chitin synthases was inviable; this was confirmed by constructing a triple mutant rescued by a plasmid carrying a CHS2 gene under control of a GAL1 promoter.
Journal ArticleDOI

Architecture of the yeast cell wall. The linkage between chitin and beta(1-->3)-glucan.

TL;DR: Experiments with appropriate mutants showed that synthesis of the chitin combined with glucan is catalyzed by chit in synthetase 3, implying that chitInase can hydrolyze the linkage between N-acetylglucosamine and glucose.
Journal ArticleDOI

Genetics and molecular biology of chitin synthesis in fungi.

TL;DR: The available data suggest that many fungi have more than one chitin synthase and that these synthases are related to the S. cerevisiae CHS and CSD gene products.
References
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Journal ArticleDOI

A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.

TL;DR: A series of yeast shuttle vectors and host strains has been created to allow more efficient manipulation of DNA in Saccharomyces cerevisiae to perform most standard DNA manipulations in the same plasmid that is introduced into yeast.
Book ChapterDOI

Fluorescence microscopy methods for yeast.

TL;DR: This chapter reviews and provides detailed protocols for the application of immunofluorescence and other fluorescence-microscopic procedures to yeast and shows that these methods are effective with other yeasts such as Schizosaccharomyces pombe and Candida albicans.
Journal ArticleDOI

Phenotypic analysis of temperature-sensitive yeast actin mutants

TL;DR: Results implicate actin in the organization and polarized growth of the yeast cell surface as well as osmotic sensitivity in conditional-lethal actin mutants.
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