- Transcriptional Analysis and Pap1-Dependence of the Unique Gene Encoding Thioredoxin Reductase from the Fission Yeast
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Hyun-Jung Kang , Sung-Min Hong , Byung-Chul Kim , Kyunghoon Kim , Eun-Hee Park , Chang-Jin Lim
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J. Microbiol. 2006;44(1):35-41.
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DOI: https://doi.org/2339 [pii]
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 Abstract
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The unique gene encoding thioredoxin reductase (TrxR) was previously cloned and characterized
from the fission yeast Schizosaccharomyces pombe, and its expression was induced by oxidative
stress. To elucidate the regulatory mechanism of the S. pombe TrxR gene, three fusion plasmids
were generated using polymerase chain reaction: pYUTR20, pYUTR30, and pYUTR40. Plasmid
pYUTR20 has an upstream region of 891 base pairs, pYUTR30 has 499 in this region, and
pYUTR40 has an 186 bp upstream region. Negatively acting sequence is located between ‒1,526
~ ‒891 bp upstream of the gene. The upstream sequence, responsible for the induction of TrxR
by menadione (MD), is situated on the ‒499 ~ ‒186 bp region, which is also required for TrxR
induction by mercuric chloride. The same region also appeared to be required for Pap1-mediated
transcriptional regulation of the TrxR gene, which contains the two plausible Pap1 binding sites,
TTACGAAT and TTACGCGA. Consistently, basal and inducible expression of the TrxR gene
was markedly lower in the Pap1-negative TP108-3C cells than in wild-type yeast cells. In summary,
up-regulation of the S. pombe TrxR gene is mediated by Pap1 via the transcriptional motif(
s) located on the ‒499 ~ ‒186 bp region.
- The Schizosaccharomyces pombe Gene Encoding [gamma]-Glutamyl Transpeptidase I Is Regulated by Non-fermentable Carbon Sources and Nitrogen Starvation
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Hong-Gyum Kim , Hey-Jung Park , Hyun-Jung Kang , Hye-Won Lim , Kyunghoon Kim , Eun-Hee Park , Kisup Ahn , Chang-Jin Lim
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J. Microbiol. 2005;43(1):44-48.
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DOI: https://doi.org/2139 [pii]
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Abstract
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In our previous study, the first structural gene (GGTI) encoding g-glutamyl transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of [beta]-galactosidase from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.
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