Transcriptional analysis and pap1-dependence of the unique gene encoding thioredoxin reductase from the fission yeast.
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 approximately -891 bp upstream of the gene. The upstream sequence, responsible for the induction of TrxR by menadione (MD), is situated on the -499 approximately -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 approximately -186 bp region.
Kang
Hyun-Jung
HJ
Division of Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon 200-701, Republic of Korea.
Hong
Sung-Min
SM
Kim
Byung-Chul
BC
Kim
Kyunghoon
K
Park
Eun-Hee
EH
Lim
Chang-Jin
CJ
D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D007763:Lac Operon; D000073718:Pancreatitis-Associated Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D050976:Basic-Leucine Zipper Transcription Factors; D000073718:Pancreatitis-Associated Proteins; C105096:Pap1 protein, S pombe; C000612141:REG3A protein, human; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D013880:Thioredoxin-Disulfide Reductase