Dolichol phosphate mannose synthase from the filamentous fungus Trichoderma reesei belongs to the human and Schizosaccharomyces pombe class of the enzyme

Joanna S. Kruszewska, Markku Saloheimo, Andrzej Migdalski, Peter Orlean, Merja Penttilä, Grazyna Palamarczyk

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30 Citations (Scopus)

Abstract

Dolichol phosphate mannose (DPM) synthase activity, which is required in N-glycosylation, O-mannosylation, and glycosylphosphatidylinositol membrane anchoring of protein, has been postulated to regulate the Trichoderma reesei secretory pathway. We have cloned a T.reesei cDNA that encodes a 243 amino acid protein whose amino acid sequence shows 67% and 65% identity, respectively, to the Schizosaccharomyces pombe and human DPM synthases, and which lacks the COOH-terminal hydrophobic domain characteristic of the Saccharomyces cerevisiae class of synthase. The Trichoderma dpm1 (Trdpm1) gene complements a lethal null mutation in the S.pombe dpm1+ gene, but neither restores viability of a S.cerevisiae dpm1-disruptant nor complements the temperature-sensitivity of the S.cerevisiae dpm1-6 mutant. The T.reesei DPM synthase is therefore a member of the ″human″ class of enzyme. Over-expression of Trdpm1 in a dpm1+::his7/dpm1+ S.pombe diploid resulted in a 4-fold increase in specific DPM synthase activity. However, neither the wild type T.reesei DPM synthase, nor a chimera consisting of this protein and the hydrophobic COOH terminus of the S.cerevisiae DPM synthase, complemented an S.cerevisiae dpm1 null mutant or gave active enzyme when expressed in E.coli. The level of the Trdpm1 mRNA in T.reesei QM9414 strain was dependent on the composition of the culture medium. Expression levels of Trdpm1 were directly correlated with the protein secretory capacity of the fungus.

Original languageEnglish
Pages (from-to)983-991
Number of pages9
JournalGlycobiology
Volume10
Issue number10
DOIs
Publication statusPublished - 1 Jan 2000
MoE publication typeA1 Journal article-refereed

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Keywords

  • Dolichol phosphate mannose synthase
  • Dpml gene
  • Trichoderma reesei

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