Protein production and secretion in an Aspergillus nidulans mutant impaired in glycosylation

• Autorzy: Perlinska-Lenart U. , Kurzatkowski W. , Janas P. , Kopinska A. , Palamarczyk G. , Kruszewska J.S.
• Języki publikacji: EN

Abstrakty

EN

O-glycosylation has been considered a limiting factor in protein secretion in filamentous fungi. Overexpression of the yeast DPM1 gene encoding dolichylphosphate mannose synthase (DPMS) in an Aspergillus nidulans mutant (BWB26A) deficient in O-glycosylation caused an increase in the number of secretory vesicles and changes in protein secretion. However, the secretory proteins, primarily O-mannosylated glucoamylase and N-glycosylated invertase, were mainly trapped in the periplasmic space. Different glycoforms of invertase were found insite the cells, in the periplasmic space and in the cultivation medium. Our data point to the importance of the cell wall as a barrier in protein secretion.

Słowa kluczowe

EN

dolichyl phosphate mannose synthase; Aspergillus nidulans; protein production; protein secretion; DPM1 gene; glycosylation; filamentous fungi; mutant

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2005
• Tom: 52
• Numer: 1
• Opis fizyczny: p.195-205,fig.,ref.

Twórcy

autor

Perlinska-Lenart U.

• Polish Academy of Sciences, A.Pawinskiego 5A, 02-106 Warsaw, Poland

autor

Kurzatkowski W.

autor

Janas P.

autor

Kopinska A.

autor

Palamarczyk G.

autor

Kruszewska J.S.

Bibliografia

• Agaphonov MO, Packeiser AN, Chechenova MB, Choi E, Ter-Avanesyan MD. (2001) Mutation of the homologue of GDP-mannose pyrophosphorylase alters cell wall structure, protein glycosylation and secretion in Hansenula polymorpha. Yeast.; 18: 391–402.
• Chomczynski P, Sacchi N. (1987) Single-step method of RNA isolation by acid guanidininium thiocyanate-phenol-chloroform extraction. Anal Biochem.; 162: 156–9.
• Gems DH, Clutterbuck AJ. (1993) Co-transformation with autonomously replicating helper plasmids facilitates gene cloning from an Aspergillus nidulans gene library. Curr Genet.; 24: 520–4.
• Gems DH, Johnstone IL, Clutterbuck AJ. (1991) An autonomously replicating plasmid transformsAspergillus nidulans at high frequency. Gene.; 98: 61–7.
• Gruber F, Visser GFJ, Kubicek CP, de Graff LH. (1990) The development of heterologous transformation system for the cellulolytic fungus Trichoderma reesei based on a pyrG-negative mutant strain. Curr Gen.; 18: 71–6.
• Kruszewska J, Palamarczyk G, Kubicek CP. (1990) Stimulation of exoprotein secretion by choline and Tween 80 in Trichoderma reesei QM 9414 correlates with increased activity of dolichol phosphate mannose synthase. J Gen Microbiol.; 136: 1293–8.
• Kruszewska J, Palamarczyk G, Kubicek CP. (1991) Mannosyl-phospho-dolichol synthase from Trichoderma reesei is activated by protein kinase dependent phosphorylation in vitro. FEMS Microbiol Lett.; 80: 81–6.
• Kruszewska J, Butterweck AH, Kurzątkowski W, Migdalski A, Kubicek CP, Palamarczyk G. (1999) Overexpression of the Saccharmyces cerevisiae mannosylphosphodolichol synthase — encoding gene in Trichoderma reesei results in an increased level of protein secretion and abnormal cell ultrastructure. Appl Environm Microbiol.; 65: 2382–7.
• Kruszewska JS, Saloheimo M, Migdalski A, Orlean P, Penttilä M, Palamarczyk G. (2000) Dolichol phosphate mannose synthase from the filamentous fungus Trichoderma reesei belongs to the human and Schizosaccharomyces pombe class of the enzyme. Glycobiology.; 10: 983–91.
• Kubicek CP, Panda T, Schreferl-Kunar G, Messner R, Gruber F. (1987) O-linked — but not N-linked-glycosylation is necessary for secretion of endoglucanase I and II by Trichoderma reesei. Can J Microbiol.; 33: 698–703.
• Kurzątkowski W, Palissa H, Liempt H, Van Doehren H, Von Kleinkauf H, Wolf WP,
• Kurylowicz W. (1991) Localisation of isopenicillin N synthase in Penicillium chrysogenum PQ 96. Apll Microbiol Biotechnol.; 35: 517–20.
• Lenart U, Kruszewska J, Or³owski J, ZdebskaE, Laudy A, Kurzątkowski W, Palamarczyk G. (2003) Alteration of Trichoderma reesei cell wall and morphology exerted by overexpression of the yeast dolichyl phosphate mannose synthase gene. II International Conference on Molecular Mechanisms of Fungal Cell Wall Biogenesis. Salamanca, Spain p. 13.
• Lovrien RE, Gusek T, Hart B. (1985) Cellulase and protease specific activities of commercially available cellulase preparations. J Appl Biochem.; 7: 258–72.
• Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. (1951) Protein measurement with the Folin phenol reagent. J Biol Chem.; 193: 265–75.
• Miller GL. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal Chem.; 31: 426–8.
• Pless DD, Palamarczyk G. (1987) Comparison of polyprenol derivatives in yeast glycosyl transfer reactions. Biochem Biophys Acta.; 529: 21–8.
• Pontercorvo G, Roper JA, Hemmons LM, Mac- Donald KD, Buffon AWJ. (1953) The genetics of Aspergillus nidulans. Adv Genet.; 5: 141–238.
• Sambrook J, Fritsch EF, Maniatis T. (1989) Molecular Cloning: a Laboratory Manual, 2nd edn, vol 1, pp 7.37–7.52. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
• Vainstein LM, Peberdy J. (1991) Regulation of invertase in Aspergillus nidulans: the effect of different carbon sources. J Gen Microbiol.; 137: 315–51.
• Yamasaki Y, Suzuki Y, Ozawa J. (1977) Purification and properties of two forms of glucoamylase from Penicillum oxalicum. Agr Biol Chem.; 41: 755–62.
• Zakrzewska A, Palamarczyk G, Krotkiewski H, Zdebska E, Saloheimo M, Penttilä M,
• Kruszewska JS. (2003) Overexpression of the gene encoding GTP-mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei. Appl Environm Microbiol.; 69: 4383–9.