Characterization, localization and functional analysis of Gpr1p, a protein affecting sensitivity to acetic acid in the yeast Yarrowia lipolytica.
Adaptation of cells to acetic acid requires a hitherto unknown number of proteins. Studies on the GPR1 gene and its encoded protein in the ascomycetous fungus Yarrowia lipolytica have revealed an involvement of this protein in the molecular processes of adaptation to acetic acid. Gpr1p belongs to a novel family of conserved proteins in prokaryotic and eukaryotic organisms that is characterized by the two motifs (A/G)NPAPLGL and SYG(X)FW (GPR1_FUN34_YaaH protein family). Analysis of four trans-dominant mutations and N-terminal deletion analysis of Gpr1p identified the amino acid sequence FGGTLN important for function of this protein in Y. lipolytica. Deletion of GPR1 slowed down adaptation to acetic acid, but had no effect on growth in the presence of acetic acid. Expression of GPR1 is induced by acetic acid and moderately repressed by glucose. It was shown by subcellular fractionation that Gpr1p is an integral membrane protein, which is also suggested by the presence of five to six putative transmembrane spanning regions. Fluorescence microscopy confirmed a localization to the plasma membrane. A model is presented describing a hypothetical function of Gpr1p during adaptation to acetic acid.
Augstein
Antje
A
Institute of Microbiology, Dresden University of Technology, Mommsenstrasse 13, D-01062 Dresden, Germany.
Barth
Kathrin
K
Institute of Microbiology, Dresden University of Technology, Mommsenstrasse 13, D-01062 Dresden, Germany.
Gentsch
Marcus
M
Institute of Microbiology, Dresden University of Technology, Mommsenstrasse 13, D-01062 Dresden, Germany.
Kohlwein
Sepp D
SD
Department of Molecular Biology, Biochemistry and Microbiology, SFB Biomembrane Research Center, University Graz, Schubertstrasse 1, A-8010 Graz, Austria.
Barth
Gerold
G
Institute of Microbiology, Dresden University of Technology, Mommsenstrasse 13, D-01062 Dresden, Germany.
D019342:Acetic Acid / Q000494:pharmacology*; D000222:Adaptation, Physiological; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001483:Base Sequence; D002462:Cell Membrane / Q000378:metabolism*; D003001:Cloning, Molecular; D015966:Gene Expression Regulation, Fungal; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D025062:Yarrowia / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism
C109334:GPR1 protein, S cerevisiae; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D019342:Acetic Acid