Budding Yeast Saccharomyces Cerevisiae as a Model Genetic Organism

Abstract

Budding yeast has served as an experimental organism for genetic research for over 50 years. The yeast shares a common cell division cycle and cellular architecture with other eukaryotes, and as a microorganism, it is easily propagated and manipulated in the laboratory. An intense focus on the central dogma, the cell cycle and sexual reproduction unleashed new fields including gene silencing, homologous recombination and differential gene expression, among others. The ease of genetic analysis allowed researchers to study processes to a degree not seen for other model organisms. As was often the case, new techniques were developed in yeast that are now broadly used. Budding yeast was the first eukaryote to be sequenced, which, in turn, led to genome‐wide analyses to map gene networks common to all life. Research on yeast has also informed us on the molecular basis of human diseases from birth defects to neurodegenerative disorders.

Key Concepts

  • Budding yeast is nonpathogenic, easy to grow and amenable to genetic analysis.
  • Core cellular functions and cell architecture are highly conserved with other eukaryotes.
  • Genes from other species can be expressed in yeast and function in place of the homologous yeast gene.
  • Sexual reproduction allows for genetic recombination and recovery of haploid yeasts expressing recessive phenotypes.
  • Homologous recombination is used to facilitate genome engineering.
  • Many tools used in molecular biology were developed in yeast (e.g. yeast two‐hybrid assay).
  • Budding yeast was the first eukaryote to have its entire genome sequenced.
  • A vast amount of yeast information has been catalogued in the Saccharomyces Genome Database (SGD) and is readily accessible to anyone.
  • The yeast knockout collection has been used for genome‐wide studies of gene function.

Keywords: Saccharomyces cerevisiae; budding yeast; homologous recombination; mating type; gene knockout; tetrad analysis; synthetic lethality; autonomously replicating sequence (ARS); yeast artificial chromosome (YAC); SaccharomycesGenome Database (SGD)

Figure 1. The life cycle of Saccharomyces cerevisiae. Cells can be grown vegetatively as haploids or diploid. Sporulation is a cellular pathway that is induced by limited nutrients. The output of sporulation is four haploid spores (tetrad) that is encased in an ascus coat.
Figure 2. Schematic of genome engineering strategies using homologous recombination. YFG is ‘Your Favorite Gene’ and URA3 is used as a selectable marker. (a) Two‐step allele replacement to replace the wild‐type allele of YFG1 with a mutated allele. For the first step, a strain with the wild‐type YFG1 allele is transformed with a plasmid bearing the mutated Y*G1 allele and the URA3 selectable marker. Transformed cells that have integrated the plasmid DNA (deoxyribonucleic acid) sequences into the chromosome by homologous recombination can be selected by growth on media lacking uracil. For the second step, a recombination event between homologous regions will loop out the duplicated sequences and depending on the placement of the recombination event, the mutation will remain in the chromosome. Although this excision event is rare, the relevant strain can be isolated through negative selection against the presence of URA3 by plating on media containing 5‐FOA (5‐fluorouracil‐6‐carboxylic acid monohydrate; 5‐fluororotic acid). Cells containing URA3 will die as the Ura3 enzyme will convert 5‐FOA to a product that is toxic to the cell. (b) Allele replacement using a linear DNA fragment with a selectable marker flanked by regions of homology that target integration in place of the wild‐type allele. Note that a region of YFG1 is missing in this fragment. (c) Gene conversion to copy information from the chromosome to a plasmid. Before transformation, a plasmid is cut with restriction enzymes to create a gap in the YFG1 sequence. This gap is filled with the wild‐type sequences in the chromosome by homologous recombination. Note that this plasmid has both CEN and ARS sequences so that it can be stably propagated in the cell.
Figure 3. Spore clones of dissected tetrads from a wild‐type strain on nutrient‐rich media containing yeast extract, peptone and dextrose (YPD) and grown for 2 days at 30 °C. Each colony arose from a single spore of a tetrad. The middle line of cells represents the pool of cells that was used to find tetrads for microdissection.
Figure 4. Meiotic segregation in the budding yeast. (a) Cross between two mutants, yfg1 × yfg2. The white and black boxes represent the mutant alleles of yfg1 and yfg2, respectively. The black circle represents the centromere. (b) Schematic of chromosomes after DNA replication and the pairing of homologous chromosomes. (c) Schematic of three possible chromosome configurations after recombination and synapsis. CO stands for crossover. (d) Tetrad classes resulting from a given crossover event are represented with the genotype indicated in the spore. Not shown: two‐strand double crossovers result in PD (parental ditype); three‐strand double crossovers result in TT (tetratype).
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Burgess, Sean M, Powers, Ted, and Mell, Joshua Chang(Nov 2017) Budding Yeast Saccharomyces Cerevisiae as a Model Genetic Organism. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0000821.pub2]