Mutations at different sites in members of the Gpr1/Fun34/YaaH protein family cause hypersensitivity to acetic acid in Saccharomyces cerevisiae as well as in Yarrowia lipolytica.
The Gpr1 protein of the ascomycetous yeast Yarrowia lipolytica belongs to the poorly characterized Gpr1/Fun34/YaaH protein family, members of which have thus far only been found in prokaryotes and lower eukaryotes. Trans-dominant mutations in the GPR1 gene result in acetic acid sensitivity of cells at low pH. Moreover, Gpr1p is subjected to phosphorylation at serine-37 in a carbon source-dependent manner. Here we show that several mutations within the ORFs of the GPR1 orthologues of Saccharomyces cerevisiae, YCR010c (ATO1) and YNR002c (ATO2), also trans-dominantly induce acetic acid hypersensitivity in this yeast. We demonstrate that the C-termini of mutated Gpr1p, Ycr010cp and Ynr002cp are necessary for the triggering of acetic acid sensitivity. Phosphorylation of Y. lipolytica Gpr1p was also affected by several mutations. Data further suggest that Gpr1p exists in an oligomeric state.
Gentsch
Marcus
M
Department of Pediatrics, University Clinic Carl Gustav Carus, Dresden, Germany.
Kuschel
Margret
M
Schlegel
Susan
S
Barth
Gerold
G
D019342:Acetic Acid / Q000378:metabolism / Q000494:pharmacology*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D015153:Blotting, Western; D004271:DNA, Fungal / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D025062:Yarrowia / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
D004271:DNA, Fungal; C109334:GPR1 protein, S cerevisiae; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D019342:Acetic Acid