title: Fluidization of membrane lipids enhances the tolerance of Saccharomyces cerevisiae to freezing and salt stress.
abstract: Unsaturated fatty acids play an essential role in the biophysical characteristics of cell membranes and determine the proper function of membrane-attached proteins. Thus, the ability of cells to alter the degree of unsaturation in their membranes is an important factor in cellular acclimatization to environmental conditions. Many eukaryotic organisms can synthesize dienoic fatty acids, but Saccharomyces cerevisiae can introduce only a single double bond at the Delta(9) position. We expressed two sunflower (Helianthus annuus) oleate Delta(12) desaturases encoded by FAD2-1 and FAD2-3 in yeast cells of the wild-type W303-1A strain (trp1) and analyzed their effects on growth and stress tolerance. Production of the heterologous desaturases increased the content of dienoic fatty acids, especially 18:2Delta(9,12), the unsaturation index, and the fluidity of the yeast membrane. The total fatty acid content remained constant, and the level of monounsaturated fatty acids decreased. Growth at 15 degrees C was reduced in the FAD2 strains, probably due to tryptophan auxotrophy, since the trp1 (TRP1) transformants that produced the sunflower desaturases grew as well as the control strain did. Our results suggest that changes in the fluidity of the lipid bilayer affect tryptophan uptake and/or the correct targeting of tryptophan transporters. The expression of the sunflower desaturases, in either Trp(+) or Trp(-) strains, increased NaCl tolerance. Production of dienoic fatty acids increased the tolerance to freezing of wild-type cells preincubated at 30 degrees C or 15 degrees C. Thus, membrane fluidity is an essential determinant of stress resistance in S. cerevisiae, and engineering of membrane lipids has the potential to be a useful tool of increasing the tolerance to freezing in industrial strains.
authors.affiliation: Department of Biotechnology, Instituto de Agroquímica y Tecnología de los Alimentos, Consejo Superior de Investigaciones Científicas, E-46100 Burjassot, Valencia, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Rodríguez-Vargas
authors.forename: Alicia
authors.initials: A
authors.lastname: Sánchez-García
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Martínez-Rivas
authors.forename: Jose Antonio
authors.initials: JA
authors.lastname: Prieto
authors.forename: Francisca
authors.initials: F
authors.lastname: Randez-Gil
mesh_terms: D001709:Biotechnology / Q000379:methods; D002463:Cell Membrane Permeability; D044943:Fatty Acid Desaturases / Q000235:genetics* / Q000378:metabolism; D005615:Freezing; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering / Q000379:methods; D018869:Heat-Shock Response; D006368:Helianthus / Q000201:enzymology / Q000235:genetics; D008563:Membrane Lipids / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D012965:Sodium Chloride / Q000494:pharmacology*
chemical_list: D008563:Membrane Lipids; D012965:Sodium Chloride; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase