1887

Microbiology Society logo

Volume 150, Issue 6

Molecular characterization of protein -mannosyltransferase and its involvement in cell-wall synthesis in Free

Abstract

Protein -glycosylation is essential for protein modification and plays important roles in eukaryotic cells. -Mannosylation of proteins occurs in the filamentous fungus . The structure and function of the gene, encoding protein --mannosyltransferase, which is responsible for the initial -mannosylation reaction in , was characterized. Disruption of the gene resulted in the reduction of protein --mannosyltransferase activity to 6 % of that of the wild-type strain and led to underglycosylation of an extracellular glucoamylase. The disruptant exhibited abnormal cell morphology and alteration in carbohydrate composition, particularly reduction in the skeletal polysaccharides in the cell wall. The results indicate that PmtA is required for the formation of a normal cell wall in .

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Dol-P, dolichol phosphate , GAI, glucoamylase I , MNT, mannosyltransferase and PMT, protein O-d-mannosyltransferase
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27005-0
2004-06-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/6/mic1501973.html?itemId=/content/journal/micro/10.1099/mic.0.27005-0&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
     [Google Scholar]
  2. Aramayo R., Adams T. H., Timberlake W. E. 1989; A large cluster of highly expressed genes is dispensable for growth and development in Aspergillus nidulans. Genetics 122:65–71
     [Google Scholar]
  3. Barratt R. W., Johnson G. B., Ogata W. N. 1965; Wild-type and mutant stocks of Aspergillus nidulans. Genetics 52:233–246
     [Google Scholar]
  4. Borgia P. T., Dodge C. L. 1992; Characterization of Aspergillus nidulans mutants deficient in cell wall chitin or glucan. J Bacteriol 174:377–383
     [Google Scholar]
  5. Borgia P. T., Iartchouk N., Riggle P. J., Winter K. R., Koltin Y., Bulawa C. E. 1996; The chsB gene of Aspergillus nidulans is necessary for normal hyphal growth and development. Fungal Genet Biol 20:193–203 [CrossRef]
     [Google Scholar]
  6. Bourdineaud J. P., van der Vaart J. M., Donzeau M., de Sampaio G., Verrips C. T., Lauquin G. J. 1998; Pmt1 mannosyl transferase is involved in cell wall incorporation of several proteins in Saccharomyces cerevisiae. Mol Microbiol 27:85–98 [CrossRef]
     [Google Scholar]
  7. Brown J. L., Bussey H. 1993; The yeast KRE9 gene encodes an O glycoprotein involved in cell surface beta-glucan assembly. Mol Cell Biol 13:6346–6356
     [Google Scholar]
  8. Brown J. L., Kossaczka Z., Jiang B., Bussey H. 1993; A mutational analysis of killer toxin resistance in Saccharomyces cerevisiae identifies new genes involved in cell wall (1-6)-beta-glucan synthesis. Genetics 133:837–849
     [Google Scholar]
  9. Burda P., Aebi M. 1999; The dolichol pathway of N-linked glycosylation. Biochim Biophys Acta 1426239–257 [CrossRef]
     [Google Scholar]
  10. Dubois M., Gilles K. A., Hamilton J. K., Rebers P. A., Smith F. 1956; Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356 [CrossRef]
     [Google Scholar]
  11. Espeso E. A., Tilburn J., Sanchez-Pulido L., Brown C. V., Valencia A. Jr, Arst H. N., Penalva M. A. 1997; Specific DNA recognition by the Aspergillus nidulans three zinc finger transcription factor PacC. J Mol Biol 274:466–480 [CrossRef]
     [Google Scholar]
  12. Farkas V. 1985; The fungal cell wall. In Fungal Protoplasts pp. 3–29 Edited by Peberdy J. F., Ferenczy L. New York: Marcel Dekker;
     [Google Scholar]
  13. Fillinger S., Ruijter G., Tamas M. K., Visser J., Thevelein J. M., d'Enfert C. 2001; Molecular and physiological characterization of the NAD-dependent glycerol 3-phosphate dehydrogenase in the filamentous fungus Aspergillus nidulans. Mol Microbiol 39:145–157 [CrossRef]
     [Google Scholar]
  14. Fontaine T., Simenel C., Dubreucq G., Adam O., Delepierre M., Lemoine J., Vorgias C. E., Diaquin M., Latge J. P. 2002; Molecular organization of the alkali-insoluble fraction of Aspergillus fumigatus cell wall. J Biol Chem 275:27594–27607
     [Google Scholar]
  15. Garcia-Rodriguez L. J., Trilla J. A., Castro C., Valdivieso M. H., Duran A., Roncero C. 2000; Characterization of the chitin biosynthesis process as a compensatory mechanism in the fks1 mutant of Saccharomyces cerevisiae. FEBS Lett 478:84–88 [CrossRef]
     [Google Scholar]
  16. Gemmill T. R., Trimble R. B. 1999; Overview of N- and O-linked oligosaccharide structures found in various yeast species. Biochim Biophys Acta 1426227–237 [CrossRef]
     [Google Scholar]
  17. Gentzsch M., Tanner W. 1996; The PMT gene family: protein O-glycosylation in Saccharomyces cerevisiae is vital. EMBO J 15:5752–5759
     [Google Scholar]
  18. Gentzsch M., Tanner W. 1997; Protein-O-glycosylation in yeast: protein-specific mannosyltransferases. Glycobiology 7:481–486 [CrossRef]
     [Google Scholar]
  19. Goto M., Ekino K., Furukawa K. 1997; Expression and functional analysis of a hyperglycosylated glucoamylase in a parental host, Aspergillus awamori var. kawachi. Appl Environ Microbiol 63:2940–2943
     [Google Scholar]
  20. Goto M., Tsukamoto M., Kwon I., Ekino K., Furukawa K. 1999; Functional analysis of O-linked oligosaccharides in threonine/serine-rich region of Aspergillus glucoamylase by expression in mannosyltransferase-disruptants of yeast. Eur J Biochem 260:596–602 [CrossRef]
     [Google Scholar]
  21. Gunnarsson A., Svensson B., Nilsson B., Svensson S. 1984; Structural studies on the O-glycosidically linked carbohydrate chains of glucoamylase G1 from Aspergillus niger. Eur J Biochem 145:463–467 [CrossRef]
     [Google Scholar]
  22. Harris S. D., Morrell J. L., Hamer J. E. 1994; Identification and characterization of Aspergillus nidulans mutants defective in cytokinesis. Genetics 136:517–532
     [Google Scholar]
  23. Harty C., Strahl S., Romisch K. 2001; O-Mannosylation protects mutant alpha-factor precursor from endoplasmic reticulum-associated degradation. Mol Biol Cell 12:1093–1101 [CrossRef]
     [Google Scholar]
  24. Haselbeck A., Tanner W. 1982; Dolichyl phosphate-mediated mannosyl transfer through liposomal membranes. Proc Natl Acad Sci U S A 79:1520–1524 [CrossRef]
     [Google Scholar]
  25. Hausler A., Ballou L., Ballou C. E., Robbins P. W. 1992; Yeast glycoprotein biosynthesis: MNT1 encodes an alpha-1,2-mannosyltransferase involved in O-glycosylation. Proc Natl Acad Sci U S A 89:6846–6850 [CrossRef]
     [Google Scholar]
  26. Hayashida S., Nakahara K., Kuroda K., Miyata T., Iwanaga S. 1989; Structure of the raw-starch-affinity site on the Aspergillus awamori var.kawachi glucoamylase I molecule. . Agric Biol Chem 53:135–141 [CrossRef]
     [Google Scholar]
  27. Horiuchi H., Fujiwara M., Yamashita S., Ohta A., Takagi M. 1999; Proliferation of intrahyphal hyphae caused by disruption of csmA, which encodes a class V chitin synthase with a myosin motor-like domain in Aspergillus nidulans. J Bacteriol 181:3721–3729
     [Google Scholar]
  28. Kellner E. M., Adams T. H. 2002; Mutations in sfdA and sfdB suppress multiple developmental mutations in Aspergillus nidulans. Genetics 160:159–168
     [Google Scholar]
  29. Lee H. H., Park J. S., Chae S. K., Maeng P. J., Park H. M. 2002; Aspergillus nidulans sod(VI)C1 mutation causes defects in cell wall biogenesis and protein secretion. FEMS Microbiol Lett 208:253–257 [CrossRef]
     [Google Scholar]
  30. Lussier M., Sdicu A. M., Bussereau F., Jacquet M., Bussey H. 1997; The Ktr1p, Ktr3p, and Kre2p/Mnt1p mannosyltransferases participate in the elaboration of yeast O- and N-linked carbohydrate chains. J Biol Chem 13:15527–15531
     [Google Scholar]
  31. Lussier M., Sdicu A. M., Bussey H. 1999; The KTR and MNN1 mannosyltransferase families of Saccharomyces cerevisiae. Biochim Biophys Acta 1426:323–334 [CrossRef]
     [Google Scholar]
  32. Martin-Blanco E., Garcia-Bellido A. 1996; Mutations in the rotated abdomen locus affect muscle development and reveal an intrinsic asymmetry in Drosophila. Proc Natl Acad Sci U S A 93:6048–6052 [CrossRef]
     [Google Scholar]
  33. Momany M., Westfall P. J., Abramowsky G. 1999; Aspergillus nidulans swo mutants show defects in polarity establishment, polarity maintenance and hyphal morphogenesis. Genetics 151:557–567
     [Google Scholar]
  34. Motoyama T., Fujiwara M., Kojima N., Horiuchi H., Ohta A., Takagi M. 1997; The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation. Mol Gen Genet 253:520–528 [CrossRef]
     [Google Scholar]
  35. Mrsa V., Tanner W. 1999; Role of NaOH-extractable cell wall proteins Ccw5p, Ccw6p, Ccw7p and Ccw8p (members of the Pir protein family) in stability of the Saccharomyces cerevisiae cell wall. Yeast 15:813–820 [CrossRef]
     [Google Scholar]
  36. Pazur J. H., Tominaga Y., Forsberg L. S., Simpson D. L. 1980; Glycoenzymes: an unusual type of glycoprotein structure for a glucoamylase. Carbohydr Res 84:103–114 [CrossRef]
     [Google Scholar]
  37. Philip B., Levin D. E. 2001; Wsc1 and Mid2 are cell surface sensors for cell wall integrity signaling that act through Rom2, a guanine nucleotide exchange factor for Rho1. Mol Cell Biol 21:271–280 [CrossRef]
     [Google Scholar]
  38. Reissig J. L., Strominger J. L., Leloir L. F. 1955; A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem 217:959–966
     [Google Scholar]
  39. Romero P. A., Lussier M., Veronneau S., Sdicu A. M., Herscovics A., Bussey H. 1999; Mnt2p and Mnt3p of Saccharomyces cerevisiae are members of the Mnn1p family of alpha-1,3-mannosyltransferases responsible for adding the terminal mannose residues of O-linked oligosaccharides. Glycobiology 9:1045–1051 [CrossRef]
     [Google Scholar]
  40. Sauer J., Sigurskjold B. W., Christensen U., Frandsen T. P., Mirgorodskaya E., Harrison M., Roepstorff P., Svensson B. 2000; Glucoamylase: structure/function relationships, and protein engineering. Biochim Biophys Acta 1543275–293 [CrossRef]
     [Google Scholar]
  41. Sharma C. B., Babczinski P., Lehle L., Tanner W. 1974; The role of dolicholmonophosphate in glycoprotein biosynthesis in Saccharomyces cerevisiae. Eur J Biochem 46:35–41 [CrossRef]
     [Google Scholar]
  42. Shaw B. D., Momany M. 2002; Aspergillus nidulans polarity mutant swoA is complemented by protein O-mannosyltransferase pmtA. Fungal Genet Biol 37:263–270 [CrossRef]
     [Google Scholar]
  43. Strahl-Bolsinger S., Immervoll T., Deutzmann R., Tanner W. 1993; PMT1, the gene for a key enzyme of protein O-glycosylation in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 90:8164–8168 [CrossRef]
     [Google Scholar]
  44. Strahl-Bolsinger S., Gentzsch M., Tanner W. 1999; Protein O-mannosylation. Biochim Biophys Acta 1426297–307 [CrossRef]
     [Google Scholar]
  45. Timpel C., Strahl-Bolsinger S., Ziegelbauer K., Ernst J. 1998; Multiple functions of Pmt1p-mediated protein O-mannosylation in the fungal pathogen Candida albicans. J Biol Chem 273:20837–20846 [CrossRef]
     [Google Scholar]
  46. Timpel C., Zink S., Strahl-Bolsinger S., Schroppel K., Ernst J. 2000; Morphogenesis, adhesive properties, and antifungal resistance depend on the Pmt6 protein mannosyltransferase in the fungal pathogen Candida albicans. J Bacteriol 182:3063–3071 [CrossRef]
     [Google Scholar]
  47. van den Hondel C. A., Punt P. J., van Gorcom R. F. 1992; Production of extracellular proteins by the filamentous fungus Aspergillus. Antonie Van Leeuwenhoek 61:153–160 [CrossRef]
     [Google Scholar]
  48. Wallis G. L., Swift R. J., Hemming F. W., Trinci A. P., Peberdy J. F. 1999; Glucoamylase overexpression and secretion in Aspergillus niger: analysis of glycosylation. Biochim Biophys Acta 1472:576–586 [CrossRef]
     [Google Scholar]
  49. Willer T., Amselgruber W., Deutzmann R., Strahl S. 2002; Characterization of POMT2, a novel member of the PMT protein O-mannosyltransferase family specifically localized to the acrosome of mammalian spermatids. Glycobiology 12:771–783 [CrossRef]
     [Google Scholar]
  50. Woo P. C., Chong K. T., Leung A. S., Wong S. S., Lau S. K., Yuen K. Y. 2003; AFLMP1 encodes an antigenic cell wall protein in Aspergillus flavus. J Clin Microbiol 41:845–850 [CrossRef]
     [Google Scholar]
  51. Yelton M. M., Hamer J. E., Timberlake W. E. 1984; Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A 81:1470–1474 [CrossRef]
     [Google Scholar]
  52. 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. Proc Natl Acad Sci U S A 91:2723–2727 [CrossRef]
     [Google Scholar]
  53. Zakrzewska A., Migdalski A., Saloheimo M., Penttila M. E., Palamarczyk G., Kruszewska J. S. 2003; cDNA encoding protein O-mannosyltransferase from the filamentous fungus Trichoderma reesei; functional equivalence to Saccharomyces cerevisiae PMT2. Curr Genet 43:11–16
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27005-0
Loading
Molecular characterization of protein O-mannosyltransferase and its involvement in cell-wall synthesis in Aspergillus nidulans
Microbiology 150, 1973 (2004); https://doi.org/10.1099/mic.0.27005-0
/content/journal/micro/10.1099/mic.0.27005-0
/content/journal/micro/10.1099/mic.0.27005-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error