title: Identification of DUOX1-dependent redox signaling through protein S-glutathionylation in airway epithelial cells.
abstract: The NADPH oxidase homolog dual oxidase 1 (DUOX1) plays an important role in innate airway epithelial responses to infection or injury, but the precise molecular mechanisms are incompletely understood and the cellular redox-sensitive targets for DUOX1-derived H2O2 have not been identified. The aim of the present study was to survey the involvement of DUOX1 in cellular redox signaling by protein S-glutathionylation, a major mode of reversible redox signaling. Using human airway epithelial H292 cells and stable transfection with DUOX1-targeted shRNA as well as primary tracheal epithelial cells from either wild-type or DUOX1-deficient mice, DUOX1 was found to be critical in ATP-stimulated transient production of H2O2 and increased protein S-glutathionylation. Using cell pre-labeling with biotin-tagged GSH and analysis of avidin-purified proteins by global proteomics, 61 S-glutathionylated proteins were identified in ATP-stimulated cells compared to 19 in untreated cells. Based on a previously established role of DUOX1 in cell migration, various redox-sensitive proteins with established roles in cytoskeletal dynamics and/or cell migration were evaluated for S-glutathionylation, indicating a critical role for DUOX1 in ATP-stimulated S-glutathionylation of β-actin, peroxiredoxin 1, the non-receptor tyrosine kinase Src, and MAPK phosphatase 1. Overall, our studies demonstrate the importance of DUOX1 in epithelial redox signaling through reversible S-glutathionylation of a range of proteins, including proteins involved in cytoskeletal regulation and MAPK signaling pathways involved in cell migration.
authors.affiliation: Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Milena
authors.initials: M
authors.lastname: Hristova
authors.affiliation: Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Carmen
authors.initials: C
authors.lastname: Veith
authors.affiliation: Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Aida
authors.initials: A
authors.lastname: Habibovic
authors.affiliation: Department of Biology, College of Arts and Sciences, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Ying-Wai
authors.initials: YW
authors.lastname: Lam
authors.affiliation: Department of Biology, College of Arts and Sciences, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Bin
authors.initials: B
authors.lastname: Deng
authors.affiliation: Department of Physiology, Faculty of Medicine, and "Lendulet" Peroxidase Enzyme Research Group, Semmelweis University, Budapest, Hungary.
authors.forename: Miklos
authors.initials: M
authors.lastname: Geiszt
authors.affiliation: Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Yvonne M W
authors.initials: YM
authors.lastname: Janssen-Heininger
authors.affiliation: Department of Pathology, Vermont Lung Center, College of Medicine, University of Vermont, Burlington, VT 05405, United States.
authors.forename: Albert
authors.initials: A
authors.lastname: van der Vliet
mesh_terms: D000818:Animals; D002465:Cell Movement; D002478:Cells, Cultured; D000074623:Dual Oxidases; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D051379:Mice; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D040901:Proteomics; D015398:Signal Transduction; D014132:Trachea / Q000166:cytology / Q000378:metabolism*
chemical_list: D000074623:Dual Oxidases; D019255:NADPH Oxidases; C431513:DUOX1 protein, human; C519130:Duox1 protein, mouse; D005978:Glutathione
keywords: Cell migration; Cysteine; DUOX1; NADPH oxidase; Proteomics; S-glutathionylation