C-terminal deletion mutant of MRE-binding transcription factor-1 inhibits MRE-driven gene expression.

Abstract

Heavy metal-induced transcriptional activation of the genes coding for metallothionein (MT) is mediated by a cis-acting DNA element, the metal-responsive element (MRE). MRE-binding transcription factor-1 (MTF-1) is a highly conserved heavy metal-induced transcriptional activator. MTF-1 also activates transcription in response to oxidative stress and regulates the expression of several cytoprotective factor genes, including MT, gamma-glutamylcysteine synthetase, and Cu/Zn-superoxide dismutase. It is thus thought that MTF-1 plays a role in cellular stress response. The physiological role of MTF-1 remains unclear because of the lack of MTF-1-specific activators and/or inhibitors. To obtain an MTF-1-specific inhibitor, we constructed an MTFDeltaC (amino acids 1-317), a C-terminal deletion mutant of MTF-1. MTFDeltaC could bind MRE and competed with MTF-1 for MTF-MRE complex formation. Transient expression of MTFDeltaC in HepG2 cells reduced MRE-driven gene expression, demonstrating that MTFDeltaC is dominant to MTF-1. HepG2 cells stably expressing MTFDeltaC showed increased susceptibility to the cytotoxic effects of tert-butyl hydroperoxide (tBH). Furthermore, we constructed Ad5MTFDeltaC, a recombinant adenovirus that expresses MTFDeltaC. Infection with the virus induced MTFDeltaC expression and increased susceptibility to the cytotoxic effects of tBH. These results indicate that MTF-1 participates in controlling the cellular redox state.