Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547

Zheng Xiang Wang, Gerald Kayingo, Anders Blomberg, Bernard A. Prior

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The dihydroxyacetone pathway, an alternative pathway for the dissimilation of glycerol via reduction by glycerol dehydrogenase and subsequent phosphorylation by dihydroxyacetone (DHA) kinase, is activated in the yeasts Saccharomyces cerevisiae and Zygosaccharomyces rouxii during osmotic stress. In experiments aimed at investigating the physiological function of the DHA pathway in Z. rouxii, a typical osmotolerant yeast, we cloned and characterized a DAK gene encoding dihydroxyacetone kinase from Z. rouxii NRRL 2547. Sequence analysis revealed a 1761 bp open reading frame, encoding a peptide composed of 587 deduced amino acids with the predicted molecular weight of 61 664 Da. As the amino acid sequence was most closely homologous (68% identity) to the S. cerevisiae Dak1p, we named the gene and protein ZrDAK1 and ZrDak1p, respectively. A putative ATP binding site was also found but no consensus element associated with osmoregulation was found in the upstream region of the ZrDAK1 gene. The ZrDAK1 gene complemented a S. cerevisiae W303-1A dak1Δdak2Δ strain by improving the growth of the mutant on 50 mmol/l dihydroxyacetone and by increasing the tolerance to dihydroxyacetone in a medium containing 5% sodium chloride, suggesting that it is a functional homologue of the S. cerevisiae DAK1. However, expression of the ZrDAK1 gene in the S. cerevisiae dak1Δdak2Δ strain had no significant effect on glycerol levels during osmotic stress. The ZrDAK1 sequence has been deposited in the public data bases under Accession No. AJ294719; regions upstream and downstream of ZrDAK1 are deposited as Accession Nos AJ294739 and AJ294720, respectively.

Original languageEnglish (US)
Pages (from-to)1447-1458
Number of pages12
JournalYeast
Volume19
Issue number16
DOIs
StatePublished - Dec 2002
Externally publishedYes

Fingerprint

Zygosaccharomyces
Gene encoding
Dihydroxyacetone
Cloning
Yeast
Saccharomyces cerevisiae
Organism Cloning
Genes
Osmotic Pressure
Glycerol
glycerol dehydrogenase
Yeasts
Osmoregulation
Amino acids
Sodium Chloride
Amino Acids
Open Reading Frames
Sequence Analysis
Phosphorylation
Amino Acid Sequence

Keywords

  • DAK1
  • DHA pathway
  • Dihydroxyacetone
  • Osmotic stress
  • Zygosaccharomyces rouxii

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547. / Wang, Zheng Xiang; Kayingo, Gerald; Blomberg, Anders; Prior, Bernard A.

In: Yeast, Vol. 19, No. 16, 12.2002, p. 1447-1458.

Research output: Contribution to journalArticle

Wang, Zheng Xiang ; Kayingo, Gerald ; Blomberg, Anders ; Prior, Bernard A. / Cloning, sequencing and characterization of a gene encoding dihydroxyacetone kinase from Zygosaccharomyces rouxii NRRL2547. In: Yeast. 2002 ; Vol. 19, No. 16. pp. 1447-1458.
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abstract = "The dihydroxyacetone pathway, an alternative pathway for the dissimilation of glycerol via reduction by glycerol dehydrogenase and subsequent phosphorylation by dihydroxyacetone (DHA) kinase, is activated in the yeasts Saccharomyces cerevisiae and Zygosaccharomyces rouxii during osmotic stress. In experiments aimed at investigating the physiological function of the DHA pathway in Z. rouxii, a typical osmotolerant yeast, we cloned and characterized a DAK gene encoding dihydroxyacetone kinase from Z. rouxii NRRL 2547. Sequence analysis revealed a 1761 bp open reading frame, encoding a peptide composed of 587 deduced amino acids with the predicted molecular weight of 61 664 Da. As the amino acid sequence was most closely homologous (68{\%} identity) to the S. cerevisiae Dak1p, we named the gene and protein ZrDAK1 and ZrDak1p, respectively. A putative ATP binding site was also found but no consensus element associated with osmoregulation was found in the upstream region of the ZrDAK1 gene. The ZrDAK1 gene complemented a S. cerevisiae W303-1A dak1Δdak2Δ strain by improving the growth of the mutant on 50 mmol/l dihydroxyacetone and by increasing the tolerance to dihydroxyacetone in a medium containing 5{\%} sodium chloride, suggesting that it is a functional homologue of the S. cerevisiae DAK1. However, expression of the ZrDAK1 gene in the S. cerevisiae dak1Δdak2Δ strain had no significant effect on glycerol levels during osmotic stress. The ZrDAK1 sequence has been deposited in the public data bases under Accession No. AJ294719; regions upstream and downstream of ZrDAK1 are deposited as Accession Nos AJ294739 and AJ294720, respectively.",
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