1887

Abstract

In this study, the identification and characterization of the homologues of -1,6-mannosyltransferases Anp1p and Och1p, designated YlAnl1p and YlOch1p, are described. In order to confirm the function of the proteins, including the previously isolated YlMnn9p, in the -glycosylation pathway, a phenotypic analysis of the disrupted strains Δ, Δ, Δ, ΔΔ and ΔΔ was performed. Disruption of the , and 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 -glycosylation process. These mutant strains, Δ, Δ, Δ, ΔΔ and ΔΔ may thus be used to establish a model for the -linked glycosylation pathway.

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2004-07-01
2020-08-15
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References

  1. Ballou C. E.. 1982; Yeast cell wall and cell surface. In The Molecular Biology of the Yeast Saccharomyces. Metabolism and Gene Expression pp.335–360Edited by Strathern J. N., Jones E. W., Broach J. R.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  2. Ballou C. E.. 1990; Isolation, characterization and properties of Saccharomyces cerevisiae mnn mutants with non-conditional glycosylation defects. Methods Enzymol185:440–470
    [Google Scholar]
  3. Ballou L., Hitzeman R. A., Lewis M. S., Ballou C. E.. 1991; Vanadate-resistant yeast mutants are defective in protein glycosylation. Prot Natl Acad Sci U S A88:3209–3212[CrossRef]
    [Google Scholar]
  4. Barth G., Gaillardin C.. 1996; Yarrowia lipolytica. In Non-Conventional Yeasts in Biotechnology: a Handbook pp.313–388Edited by Wolf K.. Berlin, Heidelberg, New York: Springer;
    [Google Scholar]
  5. Barth G., Gaillardin C.. 1997; Physiology and genetics of the dimorphic fungus Yarrowia lipolytica. FEMS Microbiol Rev18:219–237
    [Google Scholar]
  6. Beckerich J. M., Boisramé A., Gaillardin C.. 1998; Yarrowia lipolytica: a model organism for protein secretion studies. Int Microbiol1:123–130
    [Google Scholar]
  7. Boisramé A., Kabani M., Beckerich J. M., Hartmann E., Gaillardin C.. 1998; Interaction of Kar2p and Slsl1p is required for efficient co-translational translocation of secreted proteins in the yeast Yarrowia lipolytica. J Biol Chem273:30903–30908[CrossRef]
    [Google Scholar]
  8. Bui D. M., Kunze I., Wartmann T., Horstmann C., Manteuffel R., Kunze G., Förster S.. 1996; Cloning and expression of an Arxula adeninivorans glucoamylase gene in Saccharomyces cerevisiae. Appl Microbiol Biotechnol44:610–619[CrossRef]
    [Google Scholar]
  9. Casaregola S., Neuvéglise C., Lepingle A., Bon E., Artiguenave F., Wincker P., Gaillardin C.. 2000; Genomic exploration of the hemiascomycetous yeasts: 17 Yarrowia lipolytica. FEBS Lett487:95–100[CrossRef]
    [Google Scholar]
  10. Chapman R. E., Munro S.. 1994; The functioning of the yeast Golgi apparatus requires an ER protein encoded by ANP1, a member of a new family of genes affecting the secretory pathway. EMBO J13:4896–4907
    [Google Scholar]
  11. Dean N.. 1995; Yeast glycosylation mutants are sensitive to aminoglycosides. Prot Natl Acad Sci U S A92:1287–1291[CrossRef]
    [Google Scholar]
  12. Drickamer K., Taylor M. E.. 1998; Evolving views of protein glycosylation. Trends Biochem Sci23:321–324[CrossRef]
    [Google Scholar]
  13. Fields S., Song O.. 1989; A novel genetic system to detect protein–protein interactions. Nature340:245–246[CrossRef]
    [Google Scholar]
  14. Gaynor E. C., te Heesen S., Graham T. R., Aebi M., Emr S. D.. 1994; Signal-mediated retrieval of a membrane protein from the Golgi to the ER in yeast. J Cell Biol127:653–665[CrossRef]
    [Google Scholar]
  15. He F., Beckerich J. M., Gaillardin C.. 1992; A mutant of 7SL RNA in Yarrowia lipolytica affecting the synthesis of a secreted protein. J Biol Chem25:1932–1937
    [Google Scholar]
  16. Jaafar L., Leon M., Zueco J.. 2003; Identification of the MNN9 gene of Yarrowia lipolytica (YlMNN9) and phenotype analysis of a mutant ylmnn9Δ strain. Yeast20:633–644[CrossRef]
    [Google Scholar]
  17. James P., Halladay J., Craig E. A.. 1996; Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics144:1425–1436
    [Google Scholar]
  18. Jungmann J., Munro S.. 1998; Multi-protein complexes in the cis Golgi of Saccharomyces cerevisiae with α-1,6-mannosyltransferase activity. EMBO J17:423–434[CrossRef]
    [Google Scholar]
  19. Kanik-Ennulat C., Montalvo E., Neff N.. 1995; Sodium orthovanadate-resistant mutants of Saccharomyces cerevisiae show effects in Golgi-mediated protein glycosylation, sporulation, and detergent resistance. Genetics140:933–943
    [Google Scholar]
  20. Madzak C., Treton B., Blanchin-Roland S.. 2000; Strong hybrid promoters and integrative expression/secretion vectors for quasi-constitutive expression of heterologous proteins in the yeast Yarrowia lipolytica. J Mol Microbiol Biotechnol2:207–216
    [Google Scholar]
  21. Moukadiri I., Armero J., Abad A., Sentandreu R., Zueco J.. 1997; Identification of a mannoprotein present in the inner layer of the cell wall of Saccharomyces cerevisiae. J Bacteriol181:2154–2162
    [Google Scholar]
  22. Munro S.. 2001; What can yeast tell us about N-linked glycosylation in the Golgi apparatus?. FEBS Lett498:223–227[CrossRef]
    [Google Scholar]
  23. Nakanishi-Shindo N., Nakayama K., Tanaka A., Toda Y., Jigami Y.. 1993; Structure of the N-linked oligosaccharides that show the complete loss of α-1,6-polymannose outer chain fromoch1, och1 mnn1, and och1 mnn1alg3 mutants of Saccharomyces cerevisiae.. J Biol Chem268:26338–26345
    [Google Scholar]
  24. Nakayama K., Nagasu T., Shimma Y., Kuromitsu J., Jigami Y.. 1992; OCH1 encodes a novel membrane bound mannosyltransferase: outer chain elongation of asparagine-linked oligosaccharides. EMBO J11:2511–2519
    [Google Scholar]
  25. Nakayama K., Nakanishi-Shindo Y., Takana A., Haga-Toda Y., Jigami Y.. 1997; Substrate specificity of α-1,6-mannosyltransferase that initiates N-linked mannose outer chain elongation in Saccharomyces cerevisiae. FEBS Lett412:547–550[CrossRef]
    [Google Scholar]
  26. Neiman A. M., Mhaiskar V., Manus V., Galibert F., Dean N.. 1997; Saccharomyces cerevisiae HOC1, a Suppressor of pkc1, encodes a putative glycosyltransferase. Genetics145:637–645
    [Google Scholar]
  27. Parodi A. J.. 2000; Protein glycosylation and its role in protein folding. Annu Rev Biochem69:69–93[CrossRef]
    [Google Scholar]
  28. Ram A. F. J., Wolters A., ten Hoopen R., Klis F. M.. 1994; A new approach for isolating cell wall mutants in Saccharomyces cerevisiae by screening hypersensitivity to Calcofluor white. Yeast10:1019–1030[CrossRef]
    [Google Scholar]
  29. Redding K., Holcomb C., Fuller R. S.. 1991; Immunolocalization of Kex2 protease identifies a putative late Golgi compartment in the yeast Saccharomyces cerevisiae. J Cell Biol113:527–538[CrossRef]
    [Google Scholar]
  30. Romero P. A., Sleno B., Herscovics A.. 1994; Glycoprotein biosynthesis in Saccharomyces cerevisiae. Partial purification of the α-1,6-mannosyltransferase that initiates outer chain synthesis. Glycobiology4:135–140[CrossRef]
    [Google Scholar]
  31. Rothstein R.. 1983; One-step gene disruption in yeast. Methods Enzymol101:202–211
    [Google Scholar]
  32. Southard S. B., Specht C. A., Mishra C., Chen-Weiner J., Robbins P. W.. 1999; Molecular analysis of the Candida albicans homolog of Saccharomyces cerevisiae MNN9, required for glycosylation of cell wall proteins. J Bacteriol181:7439–7448
    [Google Scholar]
  33. Staden R., Beal K. F., Bonfield J. K.. 1998; Computer Methods in Molecular Biology. In Bioinformatics Methods and Protocols vol 132 pp.115–130Edited by Misener S., Krawetz S. A.. Totowa, NJ: Humana;
    [Google Scholar]
  34. Stolz J., Munro S.. 2002; The components of the Saccharomyces cerevisiae mannosyltransferase complex M-Pol I have distinct functions in mannan synthesis. J Biol Chem277:44801–44808[CrossRef]
    [Google Scholar]
  35. Swennen D., Paul M. F., Vernis L., Beckerich J. M., Fournier A., Gaillardin C.. 2002; Secretion of active anti-Ras single-chain Fv antibody by the yeasts Yarrowia lipolytica and Kluyveromyces lactis. Microbiology148:41–50
    [Google Scholar]
  36. van der Vaart J. M., Caro H. P., Chai Man J. W., Klis F. M., Verrips C. T.. 1995; Identification of three mannoproteins in the cell wall of Saccharomyces cerevisiae. J Bacteriol177:3104–3110
    [Google Scholar]
  37. Xuan J. W., Fournier P., Gaillardin C.. 1988; Cloning pf the LYS5 gene encoding saccharopine dehydrogenase from the yeast Yarrowia lipolytica by target integration. Curr Genet14:15–21[CrossRef]
    [Google Scholar]
  38. Yip C. L., Welch S. K., Klebl F., Gilbert T., Seidel P., Grant F. J., O'Hara P. J., MacKay V. L.. 1994; Cloning and analysis of the Saccharomyces cerevisiae MNN9 and MNN1 genes required for complex glycosylation of secreted proteins. Prot Natl Acad Sci U S A91:2723–2727[CrossRef]
    [Google Scholar]
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