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Creeping yeast: a simple, cheap, and robust protocol for the identification of mating type in Saccharomyces cerevisiae.

Authors
  • Arras, Samantha D M1
  • Hibbard, Taylor R2
  • Mitsugi-McHattie, Lucy1
  • Woods, Matthew A3
  • Johnson, Charlotte E1
  • Munkacsi, Andrew2
  • Denmat, Sylvie Hermann-Le1
  • Ganley, Austen R D1, 3, 4
  • 1 School of Biological Sciences, University of Auckland, Auckland, 1142, New Zealand. , (New Zealand)
  • 2 School of Biological Sciences, Victoria University of Wellington, Wellington 6012, New Zealand. , (New Zealand)
  • 3 Digital Life Institute, University of Auckland 0632, New Zealand. , (New Zealand)
  • 4 Institute of Natural and Mathematical Sciences, Massey University, Auckland 0632, New Zealand. , (New Zealand)
Type
Published Article
Journal
FEMS Yeast Research
Publisher
Oxford University Press
Publication Date
Mar 17, 2022
Identifiers
DOI: 10.1093/femsyr/foac017
PMID: 35298616
Source
Medline
Language
English
License
Unknown

Abstract

Saccharomyces cerevisiae is an exceptional genetic system, with genetic crosses facilitated by its ability to be maintained in haploid and diploid forms. Such crosses are straightforward if the mating type/ploidy of the strains are known. Several techniques can determine mating type (or ploidy), but all have limitations. Here we validate a simple, cheap and robust method to identify S. cerevisiae mating types. When cells of opposite mating type are mixed in liquid media, they 'creep' up the culture vessel sides, a phenotype that can be easily detected visually. In contrast, mixtures of the same mating type or with a diploid simply settle out. The phenotype is observable for several days under a range of routine growth conditions and with different media/strains. Microscopy suggests that cell aggregation during mating is responsible for the phenotype. Yeast knockout collection analysis identified 107 genes required for the creeping phenotype, with these being enriched for mating-specific genes. Surprisingly, the RIM101 signalling pathway was strongly represented. We propose that RIM101 signalling regulates aggregation as part of a wider, previously-unrecognized role in mating. The simplicity and robustness of this method makes it ideal for routine verification of S. cerevisiae mating type, with future studies required to verify its molecular basis. © The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.

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