Overexpression of the gene encoding GTP

mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei

Anna Zakrzewska, Grazyna Palamarczyk, Hubert Krotkiewski, Ewa Zdebska, Markku Saloheimo, Merja Penttilä, Joanna S. Kruszewska

Research output: Contribution to journalArticleScientificpeer-review

18 Citations (Scopus)

Abstract

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:α-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.

Original languageEnglish
Pages (from-to)4383-4389
Number of pages7
JournalApplied and Environmental Microbiology
Volume69
Issue number8
DOIs
Publication statusPublished - 1 Aug 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Guanosine Diphosphate Mannose
Trichoderma reesei
gene overexpression
Trichoderma
Mannose
Guanosine Triphosphate
mannose
Gross Domestic Product
Glycosylation
phosphate
phosphates
protein
gene
Genes
glycosylation
Proteins
Hypocrea
proteins
secretion
limiting factor

Cite this

@article{31002f5a6c95400695c32e64bf77aef9,
title = "Overexpression of the gene encoding GTP: mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei",
abstract = "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:α-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.",
author = "Anna Zakrzewska and Grazyna Palamarczyk and Hubert Krotkiewski and Ewa Zdebska and Markku Saloheimo and Merja Penttil{\"a} and Kruszewska, {Joanna S.}",
year = "2003",
month = "8",
day = "1",
doi = "10.1128/AEM.69.8.4383-4389.2003",
language = "English",
volume = "69",
pages = "4383--4389",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "8",

}

Overexpression of the gene encoding GTP : mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei. / Zakrzewska, Anna; Palamarczyk, Grazyna; Krotkiewski, Hubert; Zdebska, Ewa; Saloheimo, Markku; Penttilä, Merja; Kruszewska, Joanna S.

In: Applied and Environmental Microbiology, Vol. 69, No. 8, 01.08.2003, p. 4383-4389.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Overexpression of the gene encoding GTP

T2 - mannose-1-phosphate guanyltransferase, mpg1, increases cellular GDP-mannose levels and protein mannosylation in Trichoderma reesei

AU - Zakrzewska, Anna

AU - Palamarczyk, Grazyna

AU - Krotkiewski, Hubert

AU - Zdebska, Ewa

AU - Saloheimo, Markku

AU - Penttilä, Merja

AU - Kruszewska, Joanna S.

PY - 2003/8/1

Y1 - 2003/8/1

N2 - 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:α-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.

AB - 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:α-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.

UR - http://www.scopus.com/inward/record.url?scp=0042029630&partnerID=8YFLogxK

U2 - 10.1128/AEM.69.8.4383-4389.2003

DO - 10.1128/AEM.69.8.4383-4389.2003

M3 - Article

VL - 69

SP - 4383

EP - 4389

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 8

ER -