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 | |
| Publication status | Published - 1 Jan 1998 |
| MoE publication type | A1 Journal article-refereed |
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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 journal › Article › Scientific › peer-review
TY - JOUR
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.
AU - Saloheimo, Markku
AU - Penttilä, Merja
AU - Palamarczyk, Grazyna
PY - 1998/1/1
Y1 - 1998/1/1
N2 - 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.
AB - 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.
KW - dpm1 mutant
KW - Glycosylation
KW - GTP:α-D-mannose-1-phosphate guanyltransferase
KW - mpg1 gene
KW - S. cerevisiae
KW - Trichoderma reesei
UR - http://www.scopus.com/inward/record.url?scp=0031816632&partnerID=8YFLogxK
U2 - 10.1007/s002940050358
DO - 10.1007/s002940050358
M3 - Article
C2 - 9644208
AN - SCOPUS:0031816632
VL - 33
SP - 445
EP - 450
JO - Current Genetics
JF - Current Genetics
SN - 0172-8083
IS - 6
ER -