Project description:Organisms specialized to extreme environments can be the product of millions of years of evolutionary engineering and refinement. The underlying genetics can be quite distinct from those operating at earlier stages of trait innovation. In this work, we have developed the multistress-resistant yeast Kluyveromyces marxianus, which diverged from its closest relative >20 million years ago, as a model for interspecies comparative biology and genomics. In growth assays of the Kluyveromyces genus, we found that K. marxianus exhibited unique tolerance of high heat and a subset of chemical stress conditions. We then generated and analyzed omic profiles from across the genus to find molecular features associated with—and potentially causal for—K. marxianus traits. Expression profiling revealed divergent lipid processing and membrane transport programs in K. marxianus, borne out in changes in lipid utilization in experimental assays. Sequence analyses found robust evidence for expansions in gene families in the K. marxianus genome, most notably among transmembrane transporters and in metabolic enzymes. In molecular-evolution tests, we identified adaptive protein variants throughout the K. marxianus genome, among which plasma membrane transporters were over-represented. These data enable a model of the molecular mechanisms and evolutionary pressures underlying K. marxianus traits, including adaptive changes to transporters, lipid processing, and membrane functions mediating stress resistance.
Project description:Relative quantification of protein abundances of three yeast strains (Saccharomyces cerevisiae CEN.PK113-7D, Kluyveromyces marxianus CBS6556 and Yarrowia lipolytica W29) cultivate in chemostats under different conditions. The conditions for Saccharomyces cerevisiae CEN.PK113-7D are: - Standard condition – 30°C, pH 5.5 - High temperature - 36°C, pH 5.5 - Low pH - 30°C, pH 3.5 - Osmotic stress – 30°C, pH 5.5, 1M KCl The conditions for Kluyveromyces marxianus CBS6556 are: - Standard condition – 30°C, pH 5.5 - High temperature - 40°C, pH 5.5 - Low pH - 30°C, pH 3.5 - Osmotic stress – 30°C, pH 5.5, 0.6 M KCl The conditions for Yarrowia lipolytica W29 are: - Standard condition - 28°C, pH 5.5 - High temperature - 32°C, pH 5.5 - Low pH - 28°C, pH 3.5 This study is part of the OMICS data generation of CHASSY project (European Union’s Horizon 2020 grant agreement No 720824).
Project description:We have studied Schizosaccharomyces pombe, Saccharomyces cerevisiae, Scheffersomyces (Pichia) stipites, and Kluyveromyces marxianus yeast using the modified IBAQ label-free method with the spiked universal protein standard (UPS2). Each species had been prepared in 3 culturing replicates, and the similarity of the replicates was assessed using the TMT relative quantification methodology.
Project description:Protein extracts of three yeast strains (Saccharomyces cerevisiae CEN.PK113-7D, Kluyveromyces marxianus CBS6556 and Yarrowia lipolytica W29) cultivated in chemostats under different conditions. Representative samples containing aliquots of all conditions for each yeast strain were spiked with UPS2 standard (Sigma) to estimate absolute values in fmol. The conditions for Saccharomyces cerevisiae CEN.PK113-7D are: - Standard condition : 30°C, pH 5.5 - High temperature: 36°C, pH 5.5 - Low pH: 30°C, pH 3.5 - Osmotic stress : 30°C, pH 5.5, 1M KCl The conditions for Kluyveromyces marxianus CBS6556 are: - Standard condition : 30°C, pH 5.5 - High temperature: 40°C, pH 5.5 - Low pH: 30°C, pH 3.5 - Osmotic stress: 30°C, pH 5.5, 0.6 M KCl The conditions for Yarrowia lipolytica W29 are: - Standard condition: 28°C, pH 5.5 - High temperature: 32°C, pH 5.5 - Low pH: 28°C, pH 3.5 This study is part of the OMICS data generation WP of CHASSY project (European Union’s Horizon 2020 grant agreement No 720824).