Cloning, characterization and regulation of a protein disulfide isomerase from the fission yeast Schizosaccharomyces pombe

Su Jung Kim, Yeon Sook Choi, Hong Gyum Kim, Eun Hee Park, Chang Jin Lim

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5 Scopus citations

Abstract

To elucidate the physiological roles and regulation of a protein disulfide isomerase (PDI) from the fission yeast Schizosaccharomyces pombe, the full-length PDI gene was ligated into the shuttle vector pRS316, resulting in pPDI10. The determined DNA sequence carries 1,636 bp and encodes the putative 359 amino acid sequence of PDI with a molecular mass of 39,490 Da. In the amino acid sequence, the S. pombe PDI appears to be very homologous to A. thaliana PDI. The S. pombe cells harboring pPDI10 showed increased PDI activity and accelerated growth, suggesting that the cloned PDI gene is functioning and involved in the yeast growth. The 460 bp upstream region of the PDI gene was fused into promoterless β-galactosidase gene of the shuttle vector YEp367R to generate pYUPDI10. The synthesis of β-galactosidase from the PDI-lacZ fusion gene was enhanced by oxidative stress, such as superoxide anion and hydrogen peroxide. It was also induced by some non-fermentable and fermentable carbon sources. Nitrogen starvation was able to enhance the synthesis of β-galactosidase from the PDI-lacZ fusion gene. The enhancement by oxidative stress and fermentable carbon sources did not depend on the presence of Pap1. The PDI mRNA levels were increased in both Pap1-positive and Pap1-negative cells treated with glycerol. Taken together, the S. pombe PDI gene is involved in cellular growth and response to nutritional and oxidative stress.

Original languageEnglish (US)
Pages (from-to)187-196
Number of pages10
JournalMolecular Biology Reports
Volume33
Issue number3
DOIs
StatePublished - Sep 2006

Bibliographical note

Funding Information:
Acknowledgement This work was supported by a Korea Research Foundation grant (KRF-2002-070-C00062).

Keywords

  • Carbon source
  • Fission yeast
  • Hydrogen peroxide
  • LacZ fusion
  • Menadione
  • Protein disulfide isomerase
  • Schizosaccharomyces pombe
  • Transcriptional regulation

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