The thioredoxin expression of Cristaria plicata is regulated by Nrf2/ARE pathway under microcystin stimulation

Thioredoxin plays an important role in inhibiting apoptosis and protecting cells from oxidative stress. This study was aimed to clarify how the expression of Trx from Cristaria plicata is regulated by Nrf2/ARE pathway. The expression of CpTrx mRNA was significantly up-regulated in gill and kidney tissues under microcystin stress. The Nrf2 gene of Cristaria plicata was identified to possess an auto active domain bit. While CpNrf2 was knocked down by specific small RNA, CpTrx mRNA expression was significantly down-regulated. The promoter of CpTrx gene had high transcriptional activity, and this basic transcriptional activity persisted after ARE element mutation. The region of promoter −206 to +217 bp was a core promoter region and had forward regulatory elements. Gel shift Assay exhibited that the CpTrx promoter could bind to the purified proteins CpNrf2 and CpMafK in vitro. The binding phenomenon disappeared after the ARE element mutation in promoter region. Subcellular localization experiments displayed that fluorescence overlap between CpNrf2 and Trx promoter increased under microcystin toxin stress. These results suggested that Trx expression was regulated by Nrf2/ARE pathway under oxidative stress. •CpNrf2 open reading frame had self-activated domain bits ranging from 1038 bp to 1560 bp.•The promoter of the CpTrx gene continues to exhibit transcriptional activity even after ARE element has been mutated.•CpTrx promoter could bind to the purified proteins CpNrf2 and CpMafK in vitro.•Fluorescence overlap between CpNrf2 and Trx promoter increased under microcystin toxin stress.