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Applied and Environmental Microbiology, August 2003, p. 4383-4389, Vol. 69, No. 8
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.8.4383-4389.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Overexpression of the Gene Encoding GTP:Mannose-1-Phosphate Guanyltransferase, mpg1, Increases Cellular GDP-Mannose Levels and Protein Mannosylation in Trichoderma reesei

Anna Zakrzewska,1 Grazyna Palamarczyk,1 Hubert Krotkiewski,2 Ewa Zdebska,3 Markku Saloheimo,4 Merja Penttilä,4 and Joanna S. Kruszewska1*

Laboratory of Fungal Glycobiology, Institute of Biochemistry and Biophysics,1 Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences,2 Department of Biochemistry, Institute of Hematology and Blood Transfusion, Warsaw, Poland,3 VTT Biotechnology, Espoo, Finland4

Received 28 October 2002/ Accepted 13 May 2003

To elucidate the regulation and limiting factors in the glycosylation of secreted proteins, the mpg1 and dpm1 genes from Trichoderma reesei (Hypocrea jecorina) encoding GTP:{alpha}-D-mannose-1-phosphate guanyltransferase and dolichyl phosphate mannose synthase (DPMS), respectively, were overexpressed in T. reesei. No significant increases were observed in DPMS activity or protein secretion in dpm1-overexpressing transformants, whereas overexpression of mpg1 led to a twofold increase in GDP-mannose (GDPMan) levels. GDPMan was effectively utilized by mannnosyltransferases and resulted in hypermannosylation of secreted proteins in both N and O glycosylation. Overexpression of the mpg1 gene also increased the transcription of the dpm1 gene and DPMS activity. Our data indicate that the level of cellular GDPMan can play a major regulatory role in protein glycosylation in T. reesei.

* Corresponding author. Mailing address: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland. Phone: 48-22-658-44-99, x1209. Fax: 48-39-121-623. E-mail: jsk{at}ibb.waw.pl.

Applied and Environmental Microbiology, August 2003, p. 4383-4389, Vol. 69, No. 8
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.8.4383-4389.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Badejo, A. A., Tanaka, N., Esaka, M. (2008). Analysis of GDP-D-Mannose Pyrophosphorylase Gene Promoter from Acerola (Malpighia glabra) and Increase in Ascorbate Content of Transgenic Tobacco Expressing the Acerola Gene. Plant Cell Physiol 49: 126-132 [Abstract] [Full Text]  
  • Perlinska-Lenart, U., Orlowski, J., Laudy, A. E., Zdebska, E., Palamarczyk, G., Kruszewska, J. S. (2006). Glycoprotein Hypersecretion Alters the Cell Wall in Trichoderma reesei Strains Expressing the Saccharomyces cerevisiae Dolichylphosphate Mannose Synthase Gene. Appl. Environ. Microbiol. 72: 7778-7784 [Abstract] [Full Text]  


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