[Transcriptional regulation of the Hansenula polymorpha GSH2 gene in response to cadmium ion treatment].
In a previous study we cloned GSH2 gene, encoding gamma-glutamylcysteine synthetase (gammaGCS) in the yeast Hansenula polymorpha. In this study an analysis of molecular organisation of the H. polymorpha GSH2 gene promoter was conducted and the potential binding sites of Yap1, Skn7, Creb/Atf1, and Cbfl transcription factors were detected. It was established that full regulation of GSH2 gene expression in the response to cadmium and oxidative stress requires the length of GSH2 promoter to be longer than 450 bp from the start of translation initiation. To study the transcriptional regulation of H. polymorpha GSH2 gene recombinant strain, harbouring a reporter system, in which 1.832 kb regulatory region of GSH2 gene was fused to structural and terminatory regions of alcohol oxidase gene, was constructed. It was shown that maximum increase in H. polymorpha GSH2 gene transcription by 33% occurs in the rich medium under four-hour incubation with 1 microM concentration of cadmium ions. In the minimal medium the GSH2 gene expression does not correlate with the increased total cellular glutathione levels under cadmium ion treatment. We assume that the increased content of total cellular glutathione under cadmium stress in the yeast H. polymorpha probably is not controlled on the level of GSH2 gene transcription.
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Blazhenko
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Kotliarchuk
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Ubyĭvovk
D051697:Activating Transcription Factor 1 / Q000235:genetics / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002104:Cadmium / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005721:Glutamate-Cysteine Ligase / Q000235:genetics* / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D018384:Oxidative Stress; D010843:Pichia; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
D051697:Activating Transcription Factor 1; D048868:Adaptor Proteins, Signal Transducing; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D002104:Cadmium; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D005721:Glutamate-Cysteine Ligase; D005978:Glutathione