Isolation of a Trichoderma reesei cDNA encoding GTP: α-d-mannose-1-phosphate guanyltransferase involved in early steps of protein glycosylation

Joanna S. Kruszewska; Markku Saloheimo; Merja Penttilä; Grazyna Palamarczyk

doi:10.1007/s002940050358

Isolation of a Trichoderma reesei cDNA encoding GTP: α-d-mannose-1-phosphate guanyltransferase involved in early steps of protein glycosylation

Joanna S. Kruszewska, Markku Saloheimo, Merja Penttilä, Grazyna Palamarczyk^*

^*Corresponding author for this work

Polish Academy of Sciences

Research output: Contribution to journal › Article › Scientific › peer-review

17 Citations (Scopus)

Abstract

A cDNA coding for GTP:α-D-mannose-1-phosphate guanyltransferase (MPG1 transferase) (EC 2.7.7.13) was isolated from a cDNA library of the Trichoderma reesei RutC-30 strain by suppression of the yeast Saccharomyces cerevisiae mutation in the DPM1 gene encoding mannosylphosphodolichol (MPD) synthase. The nucleotide sequence of the 1.6 kb-long cDNA revealed an ORF which encodes a protein of 364 amino acids. Sequence comparisons demonstrate 70% identity with the S. cerevisiae guanyl transferase gene (MPG1) and 75% identity with the Schizosaccharomyces pombe homologue. No similarity was found with the MPD synthase encoded by the S. cerevisiae DPM1 gene. The possibility that cloned cDNA encodes a product with a MPD synthase activity was also excluded by transforming a heterozygous S. cerevisiae dpm1::LEU2/DPM1 diploid, which did not lead to the restoration of viability of the dpm1 spores. Simultaneously, a significant increase in MPG transferase activity, as compared with the wild-type yeast, was observed in cellular extracts when the mpg1 cDNA from Trichoderma was expressed in the S. cerevisiae dpm1-6 mutant.

Original language	English
Pages (from-to)	445-450
Number of pages	6
Journal	Current Genetics
Volume	33
Issue number	6
DOIs	https://doi.org/10.1007/s002940050358
Publication status	Published - 1 Jan 1998
MoE publication type	A1 Journal article-refereed

Funding

Acknowledgements We are very grateful to Dr. Peter Orlean (Univ. of Illinois at Urbana, USA) for providing yeast strains and communicating unpublished results. We thank Ms. Riitta Nurmi (VTT Biotechnology and Food Research, Espoo, Finland) for technical assistance and Dr. Andrzej Migdalski (Institute of Biochemistry and Biophysics, Warsaw, Poland) for help in sequence analysis. The work was partially financed by The Committee for Scientific Research (KBN, Poland, Grant 6P04B 01712) and the exchange program between the Finnish and Polish Academies of Science. The GenBank accession number of the T. reesei mpg1 sequence is U89991.

Keywords

dpm1 mutant
Glycosylation
GTP:α-D-mannose-1-phosphate guanyltransferase
mpg1 gene
S. cerevisiae
Trichoderma reesei

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10.1007/s002940050358

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title = "Isolation of a Trichoderma reesei cDNA encoding GTP: α-d-mannose-1-phosphate guanyltransferase involved in early steps of protein glycosylation",

abstract = "A cDNA coding for GTP:α-D-mannose-1-phosphate guanyltransferase (MPG1 transferase) (EC 2.7.7.13) was isolated from a cDNA library of the Trichoderma reesei RutC-30 strain by suppression of the yeast Saccharomyces cerevisiae mutation in the DPM1 gene encoding mannosylphosphodolichol (MPD) synthase. The nucleotide sequence of the 1.6 kb-long cDNA revealed an ORF which encodes a protein of 364 amino acids. Sequence comparisons demonstrate 70% identity with the S. cerevisiae guanyl transferase gene (MPG1) and 75% identity with the Schizosaccharomyces pombe homologue. No similarity was found with the MPD synthase encoded by the S. cerevisiae DPM1 gene. The possibility that cloned cDNA encodes a product with a MPD synthase activity was also excluded by transforming a heterozygous S. cerevisiae dpm1::LEU2/DPM1 diploid, which did not lead to the restoration of viability of the dpm1 spores. Simultaneously, a significant increase in MPG transferase activity, as compared with the wild-type yeast, was observed in cellular extracts when the mpg1 cDNA from Trichoderma was expressed in the S. cerevisiae dpm1-6 mutant.",

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Isolation of a Trichoderma reesei cDNA encoding GTP: α-d-mannose-1-phosphate guanyltransferase involved in early steps of protein glycosylation

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