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

Research output: Contribution to journalArticleScientificpeer-review

16 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 languageEnglish
Pages (from-to)445-450
Number of pages6
JournalCurrent Genetics
Volume33
Issue number6
DOIs
Publication statusPublished - 1 Jan 1998
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Guanosine Triphosphate
Glycosylation
Saccharomyces cerevisiae
Complementary DNA
Yeasts
Genes
Schizosaccharomyces
Mannose
Transferases
Spores
Diploidy
Gene Library
Open Reading Frames
mannose 1-phosphate
Amino Acids
Mutation
dolichyl-phosphate beta-D-mannosyltransferase
Proteins

Keywords

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

Cite this

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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.",
keywords = "dpm1 mutant, Glycosylation, GTP:α-D-mannose-1-phosphate guanyltransferase, mpg1 gene, S. cerevisiae, Trichoderma reesei",
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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. / Kruszewska, Joanna S.; Saloheimo, Markku; Penttilä, Merja; Palamarczyk, Grazyna.

In: Current Genetics, Vol. 33, No. 6, 01.01.1998, p. 445-450.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Isolation of a Trichoderma reesei cDNA encoding GTP

T2 - α-d-mannose-1-phosphate guanyltransferase involved in early steps of protein glycosylation

AU - Kruszewska, Joanna S.

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