Isolation of the MNN9 gene of Yarrowia lipolytica (YlMNN9) and phenotype analysis of a mutant ylmnn9 Delta strain.
In this work we describe the isolation of the Yarrowia lipolytica homologue of Saccharomyces cerevisiae MNN9 gene, which we have named YlMNN9, and the phenotype analysis of a Y. lipolytica strain containing the disrupted YlMNN9 allele. YlMNN9 was cloned using degenerate consensus oligonucleotides to generate specific probes that were in turn used to screen mini-gene libraries. The gene is defined by a 1014 bp ORF predicted to encode a protein 337 amino acids long that shares significant homology with the Mnn9ps of S. cerevisiae, Candida albicans and Hansenula polymorpha, including a putative N-terminal transmembrane domain. Disruption of YlMNN9 leads to phenotypes such as resistance to sodium orthovanadate and sensitivity to hygromycin B, compatible with a glycosylation defect, and hypersensitivity to Calcofluor white, Congo red or zymolyase, characteristic of cell wall defects. Analysis of cell wall proteins present in beta-mercaptoethanol and zymolyase extracts showed significant differences between the parental and the ylmnn9 Delta strain. These results suggest that, as has been the case with the mnn9 strain of S. cerevisiae, the ylmnn9 Delta strain we present in this work, could be used to study the cell wall proteins of Y. lipolytica and how they are organized into the cell wall.
Unidad de Microbiología, Facultad de Farmacia, Universidad de Valencia, Avda. Vicente Andres Estelles s/n, 46100-Burjassot, Valencia, Spain.
Lahcen
L
Jaafar
Maela
M
León
Jesús
J
Zueco
D000595:Amino Acid Sequence; D001483:Base Sequence; D001557:Benzenesulfonates / Q000494:pharmacology; D002176:Candida albicans / Q000235:genetics; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism; D002934:Cinnamates; D003224:Congo Red / Q000494:pharmacology; D004271:DNA, Fungal / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D005800:Genes, Fungal; D006031:Glycosylation; D006867:Hydrolases / Q000494:pharmacology; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010843:Pichia / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology; D025062:Yarrowia / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
D001557:Benzenesulfonates; D002934:Cinnamates; D004271:DNA, Fungal; D005656:Fungal Proteins; C086605:MNN9 protein, S cerevisiae; D008562:Membrane Glycoproteins; D029701:Saccharomyces cerevisiae Proteins; D012967:Sodium Dodecyl Sulfate; C037833:zymolyase; D003224:Congo Red; D014638:Vanadates; D006921:Hygromycin B; C026273:hygromycin A; C007061:C.I. Fluorescent Brightening Agent 28; D006867:Hydrolases