title: IKKα regulates human keratinocyte migration through surveillance of the redox environment.
abstract: Although the functions of H2O2in epidermal wound repair are conserved throughout evolution, the underlying signaling mechanisms are largely unknown. In this study we used human keratinocytes (HEK001) to investigate H2O2-dependent wound repair mechanisms. Scratch wounding led to H2O2production in two or three cell layers at the wound margin within ∼30 min and subsequent cysteine modification of proteins via sulfenylation. Intriguingly, exogenous H2O2treatment resulted in preferential sulfenylation of keratinocytes that adopted a migratory phenotype and detached from neighboring cells, suggesting that one of the primary functions of H2O2is to stimulate signaling factors involved in cell migration. Based on previous findings that revealed epidermal growth factor receptor (EGFR) involvement in H2O2-dependent cell migration, we analyzed oxidation of a candidate upstream target, the inhibitor of κB kinase α (IKKα; encoded byCHUK), as a mechanism of action. We show that IKKα is sulfenylated at a conserved cysteine residue in the kinase domain, which correlates with de-repression of EGF promoter activity and increased EGF expression. Thus, this indicates that IKKα promotes migration through dynamic interactions with the EGF promoter depending on the redox state within cells.
authors.affiliation: Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory, 159 Old Bar Harbor Road, Salisbury Cove, ME 04672, USA srieger@mdibl.org thomas.lisse@jax.org.|The Jackson Laboratory, Bar Harbor, ME 04609, USA.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Lisse
authors.affiliation: Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory, 159 Old Bar Harbor Road, Salisbury Cove, ME 04672, USA srieger@mdibl.org thomas.lisse@jax.org.
authors.forename: Sandra
authors.identifier: 0000-0002-9059-1670
authors.initials: S
authors.lastname: Rieger
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D003545:Cysteine / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide; D051550:I-kappa B Kinase / Q000737:chemistry / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism*; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014945:Wound Healing / Q000187:drug effects; D015027:Zebrafish
chemical_list: D004815:Epidermal Growth Factor; D006861:Hydrogen Peroxide; D051550:I-kappa B Kinase; D003545:Cysteine
keywords: EGF; H2O2; HEK001; Hydrogen peroxide; IKKα; Keratinocytes; Migration; Nuclear de-repression; Oxidation; Reactive oxygen species; Sulfenylation; Wound healing; Wound repair