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

Curr Genet. 1998 Jun;33(6):445-50. doi: 10.1007/s002940050358.

Abstract

A cDNA coding for GTP: alpha-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 DPM1gene 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Clone Cells / physiology
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification*
  • Fungal Proteins / metabolism*
  • Genetic Complementation Test
  • Glycosylation
  • Molecular Sequence Data
  • Mutation / genetics
  • Nucleotidyltransferases / genetics*
  • Nucleotidyltransferases / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Transcription Factors / genetics
  • Trichoderma / chemistry
  • Trichoderma / enzymology*
  • Trichoderma / genetics*

Substances

  • DNA, Complementary
  • Fungal Proteins
  • Transcription Factors
  • Nucleotidyltransferases
  • mannose 1-phosphate guanylyltransferase

Associated data

  • GENBANK/U89991