Journal
MOLECULAR BIOLOGY AND EVOLUTION
Volume 35, Issue 8, Pages 1823-1839Publisher
OXFORD UNIV PRESS
DOI: 10.1093/molbev/msy077
Keywords
thermotolerance; stress adaptation; experimental evolution; high-temperature growth; protein complexes; pooled segregant analysis
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Thermotolerance is a polygenic trait that contributes to cell survival and growth under unusually high temperatures. Although some genes associated with high-temperature growth (Htg(+)) have been identified, how cells accumulate mutations to achieve prolonged thermotolerance is still mysterious. Here, we conducted experimental evolution of a Saccharomyces cerevisiae laboratory strain with stepwise temperature increases for it to grow at 42 degrees C. Whole genome resequencing of 14 evolved strains and the parental strain revealed a total of 153 mutations in the evolved strains, including single nucleotide variants, small INDELs, and segmental duplication/deletion events. Some mutations persisted from an intermediate temperature to 42 degrees C, so they might be Htg(+) mutations. Functional categorization of mutations revealed enrichment of exonic mutations in the SWI/SNF complex and F-type ATPase, pointing to their involvement in high-temperature tolerance. In addition, multiple mutations were found in a general stress-associated signal transduction network consisting of Hog1 mediated pathway, RAS-cAMP pathway, and Rho1-Pkc1 mediated cell wall integrity pathway, implying that cells can achieve Htg(+) partly through modifying existing stress regulatory mechanisms. Using pooled segregant analysis of five Htg(+) phenotype-orientated pools, we inferred causative mutations for growth at 42 degrees C and identified those mutations with stronger impacts on the phenotype. Finally, we experimentally validated a number of the candidate Htg(+) mutations. This study increased our understanding of the genetic basis of yeast tolerance to high temperature.
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