trans-dominant mutations in the GPR1 gene cause high sensitivity to acetic acid and ethanol in the yeast Yarrowia lipolytica.
Acetate non-utilizing strains harbouring trans-dominant mutations in the GPR1 gene (GPR1(d)) of the dimorphic yeast Yarrowia lipolytica have been selected and characterized. These mutants are highly sensitive to low concentrations of acetic acid and ethanol, even in presence of glucose. The toxic effect of acetic acid is pH-dependent and has the strongest effect at low pH. In contrast, the action of ethanol is pH-independent. One GPR1(d) mutant has been detected that was highly sensitive to acetic acid but could still grow on ethanol, which indicates putative differences in the function of the GPR1 gene product in the sensitivity to acetic acid and ethanol. The GPR1(d) mutants exhibit a complex pleiotropic phenotype. The mutations cause changed colony morphology as well as dimorphism of cells, and induce early cell death during growth on glucose, even without the presence of dicarbon compounds. Composition of intracellular membranes, as well as morphology of vacuole and mitochondria, were strongly changed. Back-crosses with wild-type strains and analysis of recombinant strains have shown that the expression of the pleiotropic phenotype depends on the site of mutation in the GPR1 gene, as well as on the genetic background of the strain harbouring the responsive mutation. Our data suggest that Gpr1p is involved in a general response of cells to the toxic action of dicarbon compounds like acetic acid and ethanol.
Tzschoppe
K
K
Institut für Mikrobiologie, Technische Universität Dresden, Mommsenstrasse 13, D-01062 Dresden, Germany.
Augstein
A
A
Bauer
R
R
Kohlwein
S D
SD
Barth
G
G
D019342:Acetic Acid / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D000431:Ethanol / Q000494:pharmacology*; D005227:Fatty Acids / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008769:Methylnitronitrosoguanidine / Q000737:chemistry; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D016296:Mutagenesis; D010743:Phospholipids / Q000032:analysis; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D004718:Saccharomycetales / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure
D005227:Fatty Acids; C109334:GPR1 protein, S cerevisiae; D010743:Phospholipids; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D008769:Methylnitronitrosoguanidine; D000431:Ethanol; D019342:Acetic Acid