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Related Experiment Videos

Saccharomyces kluyveri mannoprotein mutants

D Pang, L Ballou, W J Zhang

    The Journal of Biological Chemistry
    |October 10, 1981
    PubMed
    Summary

    Mutant Saccharomyces kluyveri strains were identified with altered cell-wall mannoprotein carbohydrate structures. These yeast mutants exhibit defects in mannose linkage and oligosaccharide chain formation, impacting cell-wall mannoprotein composition.

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    Area of Science:

    • Biochemistry
    • Microbiology
    • Yeast Genetics

    Background:

    • Cell-surface mannoproteins are crucial for yeast cell structure and antigenicity.
    • The wild type Saccharomyces kluyveri mannoprotein has carbohydrate side chains with up to 8 mannose units.

    Purpose of the Study:

    • To identify and characterize yeast mutants with alterations in cell-wall mannoprotein carbohydrate structure.
    • To investigate the genetic basis of mannose linkage and oligosaccharide chain elongation in yeast.

    Main Methods:

    • Ethylmethane sulfonate mutagenesis of Saccharomyces kluyveri.
    • Enrichment of mutants using rabbit antiserum against wild type cells.
    • Structural analysis of mannoprotein carbohydrates using beta-elimination, acetolysis, and methylation.

    Main Results:

    • Isolated three classes of mannoprotein mutants (mnn1, mnn2, and others) with distinct carbohydrate defects.
    • The mnn1 mutant shows impaired alpha 1 leads to 3 mannose linkage, preventing chain elongation.
    • The mnn2 mutant is unable to synthesize the octasaccharide chain, while other mutants produce altered oligosaccharide ratios.

    Conclusions:

    • The identified mutations affect different genetic loci involved in yeast cell-wall mannoprotein biosynthesis.
    • These findings elucidate specific enzymatic steps in the complex process of yeast mannan assembly.

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