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

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 beta-galactosidase gene of the shuttle vector YEp367R to generate pYUPDI10. The synthesis of beta-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 beta-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.

Participation of protein disulfide isomerase 2 in the tolerance against mercury toxicity in Schizosaccharomyces pombe

“…The Aspergillus niger gene encoding PDI is also induced by ER stress (Jeenes et al, 1997). Both the S. pombe Pdi1 and Pdi2 were previously verified to participate in the control of cellular growth and response to nutritional and oxidative stresses (Kim et al, 2006;Lee et al, 2010). This work additionally demonstrates that the S.…”

“…Through the previous works (Kim et al, 2006;Lee et al, 2010), the two isoforms of PDIs were cloned and basically characterized from the fission yeast Schizosaccharomyces pombe. The first PDI, later named Pdi1, which is very homologous to Arabidopsis thaliana PDI, is involved in the yeast growth, and its expression is up-regulated by oxidative stress, such as superoxide anion and hydrogen peroxide, and nitrogen starvation in a Pap1-independent manner (Kim et al, 2006).…”

“…The first PDI, later named Pdi1, which is very homologous to Arabidopsis thaliana PDI, is involved in the yeast growth, and its expression is up-regulated by oxidative stress, such as superoxide anion and hydrogen peroxide, and nitrogen starvation in a Pap1-independent manner (Kim et al, 2006). Pdi2, a second S. pombe PDI, plays a protective role against nitrosative and nutritional stresses, and its expression is positively regulated by nitric oxide (NO) and nitrogen starvation in a Pap1-dependent manner (Lee et al, 2010).…”