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On ras gene function in yeast.

D G Fraenkel

    Proceedings of the National Academy of Sciences of the United States of America
    |July 1, 1985
    PubMed
    Summary
    This summary is machine-generated.

    A Saccharomyces cerevisiae RAS2 gene mutation impairs growth on non-carbohydrate sources and increases glycogen storage. Suppressor mutations restore growth and normalize carbohydrate reserves, suggesting RAS2

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    Area of Science:

    • Molecular Biology
    • Yeast Genetics
    • Cellular Metabolism

    Background:

    • Saccharomyces cerevisiae possesses two RAS genes, RAS1 and RAS2, involved in cellular regulation.
    • RAS genes are known to influence adenylate cyclase activity and G protein signaling pathways.
    • Cellular reserves like glycogen and trehalose are crucial for yeast survival under stress conditions.

    Purpose of the Study:

    • To investigate the function of the RAS2 gene in Saccharomyces cerevisiae.
    • To characterize the effects of a specific RAS2 mutation (ras2::LEU2) on yeast growth and metabolism.
    • To identify and analyze suppressor mutations that restore wild-type phenotypes.

    Main Methods:

    • Generating an insertion mutation in the RAS2 gene (ras2::LEU2).
    • Assessing yeast growth on various carbon sources at different pH levels.

    Related Experiment Videos

  • Quantifying glycogen and trehalose levels in wild-type and mutant strains.
  • Selecting for and characterizing spontaneous suppressor mutations.

    • Main Results:

    • The ras2::LEU2 mutation prevented growth on non-carbohydrate carbon sources, particularly at pH 7 and above.
    • Mutant strains exhibited significantly elevated glycogen levels post-glucose exhaustion.
    • Suppressor mutations restored growth on pyruvate and reduced both glycogen and trehalose to below wild-type levels.
    • Suppressors did not rescue the lethality of ras1 ras2 double mutants.

    Conclusions:

    • The RAS2 gene plays a critical role in utilizing non-carbohydrate carbon sources and regulating carbohydrate storage.
    • The observed phenotypes suggest RAS2 is involved in a G protein-mediated adenylate cyclase pathway.
    • Suppressor mutations provide insights into the complex regulatory network involving RAS genes and cellular metabolism.