title: Dual oxidase 2 generated reactive oxygen species selectively mediate the induction of mucins by epidermal growth factor in enterocytes.
abstract: Dual oxidase 2 enzyme is a member of the reactive oxygen species-generating cell membrane NADPH oxidases involved in mucosal innate immunity. It is not known if the biological activity of dual oxidase 2 is mediated by direct bacterial killing by reactive oxygen species produced by the enzyme or by the same reactive oxygen species acting as second messengers that stimulate novel gene expression. To uncover the role of reactive oxygen species and dual oxidases as signaling molecules, we have dissected the pathway triggered by epidermal growth factor to induce mucins, the principal protective components of gastrointestinal mucus. We show that dual oxidase 2 is essential for selective epidermal growth factor induction of the transmembrane MUC3 and the secreted gel-forming MUC5AC mucins. Reactive oxygen species generated by dual oxidase 2 stabilize tyrosine phosphorylation of epidermal growth factor receptor and induce MUC3 and MUC5AC through persistent activation of extracellular signal-regulated kinases 1/2-protein kinase C. Knocking down dual oxidase 2 by selective RNA targeting (siRNA) reduced epidermal growth factor receptor phosphorylation, and MUC3 and MUC5AC gene expression. Extracellular reactive oxygen species produced by dual oxidase 2, upon stimulation by epidermal growth factor, stabilize epidermal growth factor receptor phosphorylation and activate extracellular signal-regulated kinases 1/2-protein kinase C which induce MUC5AC and MUC3. Extracellular reactive oxygen species produced by dual oxidase 2 that are known to directly kill bacteria, also contribute to the maintenance of the epidermal growth factor-amplification loop, which induces mucins. These data suggest a new function of dual oxidase 2 protein in the luminal protection of the gastrointestinal tract through the induction of mucin expression by growth factors.
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: damianosalfani@libero.it.
authors.forename: Simona
authors.initials: S
authors.lastname: Damiano
authors.affiliation: Laboratorio di Ricerca Preclinica e Traslazionale, IRCCS, Centro di Riferimento Oncologico della Basilicata, Via Padre Pio, Rionero in Vulture, 85028 Potenza, Italy. Electronic address: annalisamorano@libero.it.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Morano
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: valentina.ucci@gmail.com.
authors.forename: Valentina
authors.initials: V
authors.lastname: Ucci
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: a.roberta85@libero.it.
authors.forename: Roberta
authors.initials: R
authors.lastname: Accetta
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: mondola@unina.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Mondola
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: rpaterno@unina.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Paternò
authors.affiliation: Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Italy. Electronic address: avvedim@unina.it.
authors.forename: V Enrico
authors.initials: VE
authors.lastname: Avvedimento
authors.affiliation: Dipartimento di Medicina Clinica e Chirurgia, Università di Napoli "Federico II", Via S. Pansini, 5, 80131 Naples, Italy. Electronic address: marsanti@unina.it.
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: Santillo
mesh_terms: D015153:Blotting, Western; D018938:Caco-2 Cells; D000074623:Dual Oxidases; D020895:Enterocytes / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009077:Mucins / Q000235:genetics / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D009077:Mucins; D017382:Reactive Oxygen Species; D004815:Epidermal Growth Factor; D000074623:Dual Oxidases; D019255:NADPH Oxidases; C431515:DUOX2 protein, human
keywords: DUOX; Enterocyte-like Caco-2 cells; Epidermal growth factor; Mucins; NOX; Reactive oxygen species