title: Genetic Basis of Ammonium Toxicity Resistance in a Sake Strain of Yeast: A Mendelian Case.
abstract: High concentrations of ammonium at physiological concentrations of potassium are toxic for the standard laboratory strain of Saccharomyces cerevisiae In the original description of this metabolic phenotype, we focused on the standard laboratory strains of Saccharomyces In this study, we screened a large collection of S. cerevisiae natural isolates and identified one strain that is resistant to high concentrations of ammonium. This strain, K12, was isolated in sake breweries. When the K12 strain was crossed to the standard laboratory strain (FY4), the resulting tetrads displayed 2:2 segregation of the resistance phenotype, suggesting a single gene trait. Using a bulk segregant analysis strategy, we mapped this trait to a 150-kb region on chromosome X containing the TRK1 gene. This gene encodes a transporter required for high-affinity potassium transport in S. cerevisiae Data from reciprocal hemizygosity experiments with TRK1 deletion strains in K12 and BY backgrounds, as well as analysis of the deletion of this gene in the K12 strain, demonstrate that the K12 allele of TRK1 is responsible for ammonium toxicity resistance. Furthermore, we determined the minimal amount of potassium required for both the K12 and laboratory strain needed for growth. These results demonstrate that the gene encoded by the K12 allele of TRK1 has a greater affinity for potassium than the standard allele of TRK1 found in Saccharomyces strains. We hypothesize that this greater-affinity allele of the potassium transporter reduces the flux of ammonium into the yeast cells under conditions of ammonium toxicity. These findings further refine our understanding of ammonium toxicity in yeast and provide an example of using natural variation to understand cellular processes.
authors.affiliation: Department of Genetics, Genomics and Microbiology, University of Strasbourg, CNRS, UMR7156, Strasbourg, 67083 France.
authors.forename: Cyrielle
authors.initials: C
authors.lastname: Reisser
authors.affiliation: Department of Biology, Santa Clara University, Santa Clara, California 95053.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Dick
authors.affiliation: Lewis-Sigler Institute for Integrative Genomics and Department of Ecology and Evolutionary Biology.|Howard Hughes Medical Institute.
authors.forename: Leonid
authors.initials: L
authors.lastname: Kruglyak
authors.affiliation: Lewis-Sigler Institute for Integrative Genomics and Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544.
authors.forename: David
authors.initials: D
authors.lastname: Botstein
authors.affiliation: Department of Genetics, Genomics and Microbiology, University of Strasbourg, CNRS, UMR7156, Strasbourg, 67083 France schacherer@unistra.fr hess.scu@gmail.com.
authors.forename: Joseph
authors.initials: J
authors.lastname: Schacherer
authors.affiliation: Department of Biology, Santa Clara University, Santa Clara, California 95053 schacherer@unistra.fr hess.scu@gmail.com.
authors.forename: David C
authors.initials: DC
authors.lastname: Hess
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