Stringent Mating-Type-Regulated Auxotrophy Increases the Accuracy of Systematic Genetic Interaction Screens with Saccharomyces cerevisiae Mutant Arrays

Singh I., Pass R., Togay S., Rodgers J. W., Hartman J. L.

GENETICS, vol.181, no.1, pp.289-300, 2009 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 181 Issue: 1
  • Publication Date: 2009
  • Doi Number: 10.1534/genetics.108.092981
  • Journal Name: GENETICS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.289-300
  • Bursa Uludag University Affiliated: No


A genomic collection of haploid Saccharomyces cerevisiae deletion strains provides a unique resource for systematic analysis of gene interactions. Double-mutant. haploid strains can be Constructed by the synthetic genetic array (SGA) method, wherein a query Mutation is introduced by mating to mutant arrays, selection of diploid double mutants, induction of meiosis, and selection of recombinant haploid double-mutant progeny. The mechanism of haploid selection is mating-type-regulated auxotrophy (MRA), by which prototrophy is restricted to a particular haploid genotype generated only as a result of meiosis. MRA escape leads to false-negative genetic interaction results because postmeiotic haploids that are Supposed to be under negative selection instead proliferate and mate, forming diploids that are heterozygous at interacting loci, masking phenotypes that Would be observed in a pure haploid double-mutant. Culture. This work identified factors that reduce MRA escape, including insertion of terminator and repressor sequences upstream of the MRA cassette, deletion of silent mating-type loci, and utilization of alpha-type instead of a-type MRA. Modifications engineered to reduce haploid MRA escape reduced false negative results in SGA-type analysis, resulting in > 95% sensitivity for detecting gene-gene interactions.