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Related Experiment Videos

Cell integrity and morphogenesis in a budding yeast septin mutant

Victor J Cid1, Lubica Adamíková2, Rosa Cenamor1

  • 1Departamento de Microbiología II, Facultad de Farmacia,Universidad Complutense, 28040-Madrid,Spain.

Microbiology (Reading, England)
|January 12, 1999
PubMed
Summary

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A Saccharomyces cerevisiae mutant (V327) exhibits cell lysis and abnormal cell division due to a defect in the CDC10 gene. This mutation, cdc10-11, impacts cell wall formation and sporulation, affecting Cdc10 protein localization at higher temperatures.

Area of Science:

  • Cell Biology
  • Genetics
  • Microbiology

Background:

  • The yeast Saccharomyces cerevisiae is a model organism for studying eukaryotic cell division.
  • Thermosensitive mutants are valuable tools for dissecting essential cellular processes.
  • Autolytic mutants can reveal insights into cell wall dynamics and protein secretion.

Purpose of the Study:

  • To investigate the genetic basis of a thermosensitive autolytic mutant (V327) in Saccharomyces cerevisiae.
  • To characterize the morphogenetic defect and its relationship to cell lysis and septation.
  • To identify and analyze the specific gene responsible for the observed phenotype.

Main Methods:

  • Isolation and characterization of thermosensitive autolytic mutants.
  • Transmission electron microscopy to analyze cell morphology and septation.

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  • Gene cloning by complementation of the mutant phenotype.
  • DNA sequencing to identify mutations in cloned genes.
  • Green fluorescent protein (GFP) tagging to study protein localization.

    • Main Results:

    • The V327 mutant displays efficient release of intracellular proteins and forms elongated cell chains at high temperatures.
    • Electron microscopy shows defects in septation and abnormal cell wall structure in V327.
    • Cloning and sequencing identified a point mutation in the CDC10 gene (cdc10-11) responsible for the pleiotropic phenotype.
    • The cdc10-11 mutation leads to reduced sporulation in diploid strains.
    • Cdc10-GFP localization is disrupted at the bud neck at non-permissive temperatures, correlating with the morphogenetic defect.

    Conclusions:

    • The CDC10 gene is crucial for proper cell division, cell wall integrity, and sporulation in Saccharomyces cerevisiae.
    • The cdc10-11 mutation disrupts septin function, leading to morphological defects and cell lysis.
    • Understanding Cdc10 function provides insights into the regulation of cytokinesis and cell morphogenesis.