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The Human Peroxiredoxin 6 (Prdx6) Promoter Activity is Stimulated by H 2 O 2
Author(s) -
Chowdhury Ibrul H,
Feinstein Sheldon I,
Manevich Yefim,
Mo Yiqun,
Fisher Aron B.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1457
Subject(s) - microbiology and biotechnology , transfection , promoter , gene , luciferase , plasmid , clone (java method) , genbank , transcription factor , biology , expression vector , reporter gene , chemistry , gene expression , recombinant dna , genetics
Peroxiredoxin 6 (Prdx6), the only member of the Prdx family that contains a single conserved cysteine residue, is an important lung anti‐oxidant enzyme. To study the human Prdx6 promoter, we used A549, a human lung epithelial cell line which expresses both Prdx6 and Nrf2, an oxidative stress‐responsive transcription factor. Treatment of A549 cells with 200μM H 2 O 2 for 12 h increased Prdx6 mRNA levels 2.5 fold compared with control. 1524 bp of the human Prdx6 promoter region obtained by PCR was cloned into the pSEAP2‐Basic vector (BD, Bioscience) for analysis using secreted alkaline phosphatase as a reporter gene. Promoter clone S82, obtained from BAC clone 638L8, contains a single A to T transversion at position −273 compared to the Genbank sequence. Potential antioxidant response elements (AREs) exist at positions −401 to −393 and −387 to −349. AREs have been shown to bind Nrf2. The pSEAP2‐Basic plasmid containing the promoter was transfected into A549 cells. A plasmid expressing β‐galactosidase from the SV40 promoter was used as a transfection control. We measured SEAP activity after transfection for 24 h and treated the cells with 400μM H 2 O 2 for another 8 hours. Prdx6 promoter activity increased by about 2.5 fold in the presence of H 2 O 2 . These data indicate that oxidative stress can induce an increased transcriptional response from the Prdx6 promoter. [PO1‐HL‐19737]