title: Trafficking of the transcription factor Nrf2 to promyelocytic leukemia-nuclear bodies: implications for degradation of NRF2 in the nucleus.
abstract: Ubiquitylation of Nrf2 by the Keap1-Cullin3/RING box1 (Cul3-Rbx1) E3 ubiquitin ligase complex targets Nrf2 for proteasomal degradation in the cytoplasm and is an extensively studied mechanism for regulating the cellular level of Nrf2. Although mechanistic details are lacking, reports abound that Nrf2 can also be degraded in the nucleus. Here, we demonstrate that Nrf2 is a target for sumoylation by both SUMO-1 and SUMO-2. HepG2 cells treated with As2O3, which enhances attachment of SUMO-2/3 to target proteins, increased SUMO-2/3-modification (polysumoylation) of Nrf2. We show that Nrf2 traffics, in part, to promyelocytic leukemia-nuclear bodies (PML-NBs). Cell fractions harboring key components of PML-NBs did not contain biologically active Keap1 but contained modified Nrf2 as well as RING finger protein 4 (RNF4), a poly-SUMO-specific E3 ubiquitin ligase. Overexpression of wild-type RNF4, but not the catalytically inactive mutant, decreased the steady-state levels of Nrf2, measured in the PML-NB-enriched cell fraction. The proteasome inhibitor MG-132 interfered with this decrease, resulting in elevated levels of polysumoylated Nrf2 that was also ubiquitylated. Wild-type RNF4 accelerated the half-life (t&frac12;) of Nrf2, measured in PML-NB-enriched cell fractions. These results suggest that RNF4 mediates polyubiquitylation of polysumoylated Nrf2, leading to its subsequent degradation in PML-NBs. Overall, this work identifies Nrf2 as a target for sumoylation and provides a novel mechanism for its degradation in the nucleus, independent of Keap1.
authors.affiliation: Department of Neuroscience and Pharmacology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599.
authors.forename: Melanie Theodore
authors.initials: MT
authors.lastname: Malloy
authors.affiliation: Department of Neuroscience and Pharmacology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599.
authors.forename: Deneshia J
authors.initials: DJ
authors.lastname: McIntosh
authors.affiliation: Department of Neuroscience and Pharmacology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599.
authors.forename: Treniqka S
authors.initials: TS
authors.lastname: Walters
authors.affiliation: Department of Neuroscience and Pharmacology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599.
authors.forename: Andrea
authors.initials: A
authors.lastname: Flores
authors.affiliation: Department of Biochemistry and Cancer Biology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599.
authors.forename: J Shawn
authors.initials: JS
authors.lastname: Goodwin
authors.affiliation: Department of Physiology, School of Medicine, Meharry Medical College, Nashville, Tennessee 37208-3599. Electronic address: iarinze@mmc.edu.
authors.forename: Ifeanyi J
authors.initials: IJ
authors.lastname: Arinze
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D017930:Genes, Reporter; D056945:Hep G2 Cells; D006801:Humans; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D018384:Oxidative Stress; D017434:Protein Structure, Tertiary; D058207:Sumoylation; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D009687:Nuclear Proteins; C114070:RNF4 protein, human; D014157:Transcription Factors
keywords: Degradation of Nrf2; Fluorescence Resonance Energy Transfer (FRET); Promyelocytic Leukemia-Nuclear Bodies; Protein Degradation; RNF4; Sumoylation; Trafficking; Transcription Factors
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