Identification and characterization of two alpha-1,6-mannosyltransferases, Anl1p and Och1p, in the yeast Yarrowia lipolytica.
In this study, the identification and characterization of the Yarrowia lipolytica homologues of Saccharomyces cerevisiae alpha-1,6-mannosyltransferases Anp1p and Och1p, designated YlAnl1p and YlOch1p, are described. In order to confirm the function of the Y. lipolytica proteins, including the previously isolated YlMnn9p, in the N-glycosylation pathway, a phenotypic analysis of the disrupted strains Delta Ylmnn9, Delta Ylanl1, Delta Yloch1, Delta Ylanl1 Delta Ylmnn9 and Delta Ylmnn9 Delta Yloch1 was performed. Disruption of the YlMNN9, YlANL1 and YlOCH1 genes caused an increased sensitivity to SDS, compatible with a glycosylation defect, and to Calcofluor White, characteristic of cell-wall defects. Moreover, Western-blot analysis of a heterologous glycosylated protein confirmed a direct role of YlMnn9p and YlAnl1p in the N-glycosylation process. These mutant strains, Delta Ylmnn9, Delta Ylanl1, Delta Yloch1, Delta Ylanl1 Delta Ylmnn9 and Delta Ylmnn9 Delta Yloch1 may thus be used to establish a model for the Y. lipolytica N-linked glycosylation pathway.
Barnay-Verdier
Stéphanie
S
Laboratoire de Microbiologie et Génétique Moléculaire, CNRS-Institut National Agronomique Paris-Grignon-INRA, 78850 Thiverval-Grignon, France.
Boisramé
Anita
A
Laboratoire de Microbiologie et Génétique Moléculaire, CNRS-Institut National Agronomique Paris-Grignon-INRA, 78850 Thiverval-Grignon, France.
Beckerich
Jean-Marie
JM
Laboratoire de Microbiologie et Génétique Moléculaire, CNRS-Institut National Agronomique Paris-Grignon-INRA, 78850 Thiverval-Grignon, France.
D000595:Amino Acid Sequence; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D006031:Glycosylation; D008364:Mannosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C075636:OCH1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C090211:ANP1 protein, S cerevisiae; D008364:Mannosyltransferases