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Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models
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Hypersensitivity to heavy water: a new conditional phenotype.

B Bartel1, A Varshavsky

  • 1Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

Cell
|March 25, 1988
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method using heavy water (D2O) to create conditional mutations in yeast. This D2O-sensitive (ds) mutant technique offers a novel approach for genetic studies, particularly in organisms not easily studied with temperature-sensitive methods.

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Area of Science:

  • Genetics and Molecular Biology
  • Yeast Model Organisms
  • Biochemistry

Background:

  • Wild-type yeast strains (Saccharomyces cerevisiae) exhibit growth in heavy water (D2O).
  • Conventional methods like temperature-sensitive (ts) mutations are widely used for conditional genetics.
  • Limitations exist in ts and cold-sensitive (cs) approaches for certain genes and organisms.

Purpose of the Study:

  • To develop and characterize a novel D2O-sensitive (ds) mutant screening method in yeast.
  • To assess the frequency and genetic distribution of ds mutations.
  • To evaluate the utility of the ds technique for generating conditional mutations in previously intractable genes and for broader applications in genetics.

Main Methods:

  • Chemical mutagenesis of wild-type yeast (S. cerevisiae).
  • Screening for mutants unable to grow on 90% D2O media but capable of growth on H2O media.
  • Complementation analysis to determine the distribution of ds mutations across complementation groups.
  • Phenotypic characterization of ds mutants for other conditional sensitivities (heat, cold, osmotic).

Main Results:

  • A significant frequency of D2O-sensitive (ds) mutants was obtained, comparable to temperature-sensitive (ts) mutants.
  • ds mutations were found to be distributed across numerous complementation groups.
  • Most ds mutants did not exhibit other conditional phenotypes; conversely, few cell division cycle ts mutants were also ds.

Conclusions:

  • The D2O-sensitivity (ds) technique is a valuable new method for generating conditional mutations.
  • This approach is particularly useful for genes not amenable to traditional ts or cs screening.
  • The ds technique holds potential for generating conditional mutants in homeothermic animals, including mammals and birds.