Genetic and Cytological Methods to Study ESCRT Cell Cycle Function in Fission Yeast

Methods Mol Biol. 2019:1998:239-250. doi: 10.1007/978-1-4939-9492-2_18.

Abstract

The fission yeast Schizosaccharomyces pombe, an ascomycete fungus, is an established model organism for studying eukaryotic molecular and cellular events such as the cell cycle due to its powerful genetics, a sequenced genome, and the ease of molecular manipulation (Wood et al., Nature 415:871-880, 2002; Hoffman et al., Genetics 201:403-423, 2015). This chapter describes genetic and cytological methods to study endosomal sorting complex required for transport (ESCRT) function during the cell cycle in fission yeast. These include tetrad analysis to allow the creation of double mutants to test for genetic interactions by synthetic phenotype characterization, such as cellular growth and the analysis of division septa by calcofluor-white staining.

Keywords: Anillin; Calcofluor-white; ESCRT; Fission yeast; Genetic interaction; Polo kinase; Schizosaccharomyces pombe; Septation; Synthetic phenotype; Tetrad analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Benzenesulfonates / chemistry
  • Cell Culture Techniques / methods
  • Cell Cycle*
  • Endosomal Sorting Complexes Required for Transport / physiology*
  • Fluorescent Dyes / chemistry
  • Genotyping Techniques / methods
  • Intravital Microscopy / methods*
  • Microscopy, Fluorescence / methods
  • Mutation
  • Schizosaccharomyces / physiology*
  • Schizosaccharomyces pombe Proteins / physiology*
  • Staining and Labeling / methods

Substances

  • Benzenesulfonates
  • Endosomal Sorting Complexes Required for Transport
  • Fluorescent Dyes
  • Schizosaccharomyces pombe Proteins
  • C.I. Fluorescent Brightening Agent 28