| Transcriptional regulation of rat gamma-glutamate cysteine ligase catalytic subunit gene is mediated through a distal antioxidant response element. | |
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MedLine Citation:
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PMID: 19540342 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Despite it being a quintessential Phase II detoxification gene, the transcriptional regulation of the rat gamma-glutamate cysteine ligase catalytic subunit (GCLC) is controversial. Computer-based sequence analysis identified three putative antioxidant response elements (AREs) at positions -889 to -865 (ARE1), -3170 to -3146 (ARE2) and -3901 to -3877 (ARE3) in the 5'-flanking region of the transcriptional start site. Transfections of individual ARE-luciferase reporter gene constructs into H4IIE cells, a rat hepatoma cell line, identified ARE3 as the functional promoter. Chromatin immunoprecipitation assays using primary rat hepatocytes showed that the transcription factor Nrf2, which is known to regulate ARE-mediated genes, is associated with ARE3. Co-transfection of H4IIE cells with luciferase reporter plasmids containing Gclc ARE3 and an Nrf2 expression plasmid resulted in a 3-fold activation of ARE3-mediated transcription relative to controls. "Loss-of-function" analysis for Nrf2 by small interfering RNA (siRNA) revealed that ARE3-mediated expression was significantly impaired while site-directed mutagenesis of the ARE3-luciferase reporter abolished Nrf2-mediated induction. Treatment with two known Nrf2 inducers, R-(alpha)-lipoic acid and anetholedithiolethione, showed that the inducible expression of the GCLC gene was also regulated by the ARE3 element. Taken together, these results show that Nrf2 regulates the constitutive expression of rat Gclc through a distal ARE present in its 5'-flanking region. This is the first report showing that rat Gclc is under the transcriptional control of the Nrf2-ARE pathway on a constitutive basis. |
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Authors:
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Swapna V Shenvi; Eric J Smith; Tory M Hagen |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2009-06-18 |
Journal Detail:
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Title: Pharmacological research : the official journal of the Italian Pharmacological Society Volume: 60 ISSN: 1096-1186 ISO Abbreviation: Pharmacol. Res. Publication Date: 2009 Oct |
Date Detail:
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Created Date: 2009-08-31 Completed Date: 2009-12-22 Revised Date: 2012-10-09 |
Medline Journal Info:
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Nlm Unique ID: 8907422 Medline TA: Pharmacol Res Country: England |
Other Details:
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Languages: eng Pagination: 229-36 Citation Subset: IM |
Affiliation:
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Linus Pauling Institute, Oregon State University, Corvallis, OR 97331, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Cell Line, Tumor Cells, Cultured Chromatin Immunoprecipitation Glutamate-Cysteine Ligase / genetics*, metabolism Hepatocytes / metabolism Molecular Sequence Data NF-E2-Related Factor 2 / genetics, metabolism Promoter Regions, Genetic RNA, Small Interfering / genetics Rats Transcription Initiation Site* Transcriptional Activation* Transfection |
| Grant Support | |
ID/Acronym/Agency:
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ES00240/ES/NIEHS NIH HHS; P01 AT002034/AT/NCCAM NIH HHS; P01 AT002034-030002/AT/NCCAM NIH HHS; P01AT002034-06/AT/NCCAM NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/NF-E2-Related Factor 2; 0/RNA, Small Interfering; EC 6.3.2.2/Glutamate-Cysteine Ligase; EC 6.3.2.2./GCLC protein, rat |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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