- 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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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.
- Transcriptional Regulation of the Schizosaccharomyces pombe Gene Encoding Glutathione S-Transferase I by a Transcription Factor Pap1
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Hong-Gyum Kim , Byung-Chul Kim , Kyunghoon Kim , Eun-Hee Park , Chang-Jin Lim
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J. Microbiol. 2004;42(4):353-356.
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DOI: https://doi.org/2099 [pii]
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In a previous study, a gst gene was isolated from the fission yeast Schizosaccharomyces pombe. This gene was dubbed gst I, and was characterized using the gstI-lacZ fusion plasmid pYSH2000. In this work, four additional fusion plasmids, pYSHSD1, pYSHSD2, pYSHSD3 and pYSHSD4, were constructed, in order to carry (respectively) 770, 551, 358 and 151 bp upstream regions from the translational initiation point. The sequence responsible for induction by aluminum, mercury and hydrogen peroxide was located in the range between -1,088 and -770 bp upstream of the S. pombe gst I gene. The same region was identified to contain the nucleotide sequence responsible for regulation by Pap1, and has one putative Pap1 binding site, TTACGTAT, located in the range between -954 ~ -947 bp upstream of the gst I gene. Negatively acting sequences are located between -1,088 and -151 bp. These findings imply that the Pap1 protein is involved in basal and inducible transcription of the gst I gene in the fission yeast S. pombe.
- Transcriptional Regulation of the Gene Encoding g-Glutamylcysteine Synthetase from the Fission Yeast Schizosaccharomyces pombe
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Su-Jung Kim , Hong-Gyum Kim , Byung-Chul Kim , Kyunghoon Kim , Eun-Hee Park , Chang-Jin Lim
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J. Microbiol. 2004;42(3):233-238.
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DOI: https://doi.org/2083 [pii]
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Transcriptional regulation of the Schizosaccharomyces pombe [gamma]-glutamylcysteine synthetase (GCS) gene was examined using the two GCS-lacZ fusion plasmids pUGCS101 and pUGCS102, which harbor 607 bp and 447 bp upstream regions, respectively. The negatively-acting sequence was located in the -607 ~ -447 bp upstream region of the GCS gene. The upstream sequence responsible for induction by menadione (MD) and L-buthionine-(S, R)-sulfoximine (BSO) resides in the -607 ~ -447 bp region, whereas the sequence which codes for nitric oxide induction is located within the -447 bp region, measured from the translational initiation point. Carbon source-dependent regulation of the GCS gene appeared to be dependent on the nucleotide sequence within -447 bp region. The transcription factor Pap1 is involved in the induction of the GCS gene by MD and BSO, but not by nitric oxide. Induction of the GCS gene occurring due to low glucose concentration does not depend on the presence of Pap1. These data imply that induction by MD and BSO may be mediated by the Pap1 binding site, probably located in the -607 ~ -447 region, and also that the nitric oxide-mediated regulation of the S. pombe GCS gene may share a similar mechanism with its carbon-dependent induction.
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