Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Insect Ornithine Decarboxylase (ODC) Complements SPE1 Knock-Out of Yeast Saccharomyces cerevisiae

  • Choi, Soon-Yong (Department of Biotechnology, Hannam University) ;
  • Park, Hee Yun (Department of Biological Sciences, Hannam University) ;
  • Paek, Aron (Department of Biological Sciences, Hannam University) ;
  • Kim, Gil Seob (Department of Biological Sciences, Hannam University) ;
  • Jeong, Seong Eun (Department of Biological Sciences, Hannam University)
  • Received : 2009.09.03
  • Accepted : 2009.10.19
  • Published : 2009.12.31
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Abstract

Ornithine decarboxylase (ODC) is a rate-limiting enzyme in the biosynthesis of polyamines, which are essential for cell growth, differentiation, and proliferation. This report presents the characterization of an ODC-encoding cDNA (SlitODC) isolated from a moth species, the tobacco cutworm, Spodoptera litura (Lepidoptera); its expression in a polyamine-deficient strain of yeast, S. cerevisiae; and the recovery in polyamine levels and proliferation rate with the introduction of the insect enzyme. SlitODC encodes 448 amino acid residues, 4 amino acids longer than B. mori ODC that has 71% identity, and has a longer C-terminus, consistent with B. mori ODC, than the reported dipteran enzymes. The null mutant yeast strain in the ODC gene, SPE1, showed remarkably depleted polyamine levels; in putrescine, spermidine, and spermine, the levels were > 7, > 1, and > 4%, respectively, of the levels in the wild-type strain. This consequently caused a significant arrest in cell proliferation of > 4% of the wild-type strain in polyamine-free media. The transformed strain, with the substituted SlitODC for the deleted endogenous ODC, grew and proliferated rapidly at even a higher rate than the wild-type strain. Furthermore, its polyamine content was significantly higher than even that in the wild-type strain as well as the spe1-null mutant, particularly with a very continuously enhanced putrescine level, reflecting no inhibition mechanism operating in the putrescine synthesis step by any corresponding insect ODC antizymes to SlitODC in this yeast system.

Keywords

Acknowledgement

Supported by : Hannam University, Small and Medium Enterprise Agency

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