Distinct roles of thioredoxin in the cytoplasm and in the nucleus. A two-step mechanism of redox regulation of transcription factor NF-kappaB

Abstract Oxidative stresses such as UV irradiation to mammalian cells triggers a variety of oxistress responses including activation of transcription factors. Recently, activation of nuclear factor-κB (NF-κB) has been shown to be under oxidoreduction (redox) regulation controlled by thioredoxin (TRX), which is one of major endogenous redox-regulating molecules with thiol reducing activity. In order to elucidate where in the cellular compartment TRX participates in NF-κB regulation, we investigated the intracellular localization of TRX. UVB irradiation induced translocation of TRX from the cytoplasm into the nucleus. In our in vitro diamide-induced cross-linking study, we showed that TRX can associate directly with NF-κB p50. Overexpression of wild-type TRX suppressed induction of luciferase activity under NF-κB-binding sites in response to UV irradiation compared with the mock transfectant. In contrast, overexpression of nuclear-targeted TRX enhanced the luciferase activity. Thus, TRX seems to play dual and opposing roles in the regulation of NF-κB. In the cytoplasm, it interferes with the signals to IκB kinases and blocks the degradation of IκB. In the nucleus, however, TRX enhances NF-κB transcriptional activities by enhancing its ability to bind DNA. This two-step TRX-dependent regulation of the NF-κB complex may be a novel activation mechanism of redox-sensitive transcription factors.