La Autoantigen Mediates Oxidant Induced De Novo Nrf2 Protein Translation

Nrf2 gene encodes a transcription factor that regulates the expression of a cluster of antioxidant and detoxification genes. Recent works from our laboratory indicate that oxidative stress causes rapid de novo synthesis of Nrf2 protein. We have found that 5′ Untranslated Region (5′UTR) of Nrf2 allows the mRNA to undergo an Internal Ribosomal Entry Site (IRES) mediated protein translation. Using liquid chromatography tandem MS, we have discovered that La/SSB protein bound to Nrf2 5′UTR in response to oxidative stress. In vitro RNA binding and in vivo ribonucleoprotein immunoprecipitation showed H2O2 dose and time dependent increases of La/SSB binding to Nrf2 5′UTR. La/SSB protein translocated from the nuclei to cytoplasm and distributed in the perinuclear space in cells treated with H2O2. Isolation of ribosomal fractions indicated that oxidants caused an association of La/SSB with ribosomes. Physical interaction of La/SSB with representative proteins from the small or large subunits of ribosomes was found to increase in cells responding to H2O2 treatment. Knocking down La/SSB gene with siRNA prevented Nrf2 protein elevation or Nrf2 5′UTR activation by oxidants. In contrast, overexpression of La/SSB gene was able to enhance Nrf2 5′UTR activation and Nrf2 protein increase. Our data suggest that oxidants cause nuclear export of La/SSB protein and subsequent association of La/SSB with Nrf2 5′UTR and ribosomes. These events contribute to de novo Nrf2 protein translation because of oxidative stress.