Genetic analysis of regulatory mutants affecting synthesis of extracellular proteinases in the yeast Yarrowia lipolytica: identification of a RIM101/pacC homolog.
Depending on the pH of the growth medium, the yeast Yarrowia lipolytica secretes both an acidic proteinase and an alkaline proteinase, the synthesis of which is also controlled by carbon, nitrogen, and sulfur availability, as well as by the presence of extracellular proteins. Recessive mutations at four unlinked loci, named PAL1 to PAL4, were isolated which prevent alkaline proteinase derepression under conditions of carbon and nitrogen limitation at pH 6.8. These mutations markedly affect mating and sporulation. A dominant suppressor of all four PAL mutations was isolated from a wild-type genomic library, which turned out to be a C-terminally truncated form of a 585-residue transcriptional factor of the His2Cys2 zinc finger family, which we propose to call YlRim101p. Another C-terminally truncated version of YlRim101p (419 residues) is encoded by the dominant RPH2 mutation previously isolated as expressing alkaline protease independently of the pH. YlRim101p is homologous to the transcriptional activators Rim101p of Saccharomyces cerevisiae, required for entry into meiosis, and PacC of Aspergillus nidulans and Penicillium chrysogenum, which were recently shown to mediate regulation by ambient pH. YlRim101p appears essential for mating and sporulation and for alkaline proteinase derepression. YlRIM101 expression is autoregulated, maximal at alkaline pH, and strongly impaired by PAL mutations.
Lambert
M
M
Laboratoire de Génétique Moléculaire et Cellulaire INRA-CNRS, Institut National Agronomique Paris-Grignon, Thiverval-Grignon, France.
Blanchin-Roland
S
S
Le Louedec
F
F
Lepingle
A
A
Gaillardin
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics* / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D004718:Saccharomycetales / Q000235:genetics*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics*; D013172:Spores, Fungal; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D015003:Yeasts / Q000235:genetics*
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C092394:PacC protein, Aspergillus; C489601:RIM101 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D012697:Serine Endopeptidases; C067468:Yarrowia lipolytica alkaline extracellular protease