Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-B Binding Site

Autor(es)

Frazao, Josias B. ^[1] ; Thain, Alison ^{[2, 3]} ; Zhu, Zhiqing ^{[2, 3]} ; Luengo, Marcos ^[4] ; Condino-Neto, Antonio ^[1] ; Newburger, Peter E. ^{[2, 3]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Immunol, BR-05508900 Sao Paulo, SP - Brazil

^[2] Univ Massachusetts, Sch Med, Dept Pediat, Worcester, MA 01655 - USA

^[3] Univ Massachusetts, Sch Med, Dept Mol Cellular & Canc Biol, Worcester, MA 01655 - USA

^[4] Univ Estadual Campinas, Sch Med, Ctr Invest Pediat, BR-13081970 Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Cellular Biochemistry; v. 116, n. 9, p. 2008-2017, SEP 2015.
• Citações Web of Science: 2

Resumo

The human CYBB gene encodes the gp91-phox component of the phagocyte oxidase enzyme complex, which is responsible for generating superoxide and other downstream reactive oxygen species essential to microbial killing. In the present study, we have identified by sequence analysis a putative NF-B binding site in a DNase I hypersensitive site, termed HS-II, located in the distant 5 flanking region of the CYBB gene. Electrophoretic mobility assays showed binding of the sequence element by recombinant NF-B protein p50 and by proteins in nuclear extract from the HL-60 myeloid leukemia cell line corresponding to p50 and to p50/p65 heterodimers. Chromatin immunoprecipitation demonstrated NF-B binding to the site in intact HL-60 cells. Chromosome conformation capture (3C) assays demonstrated physical interaction between the NF-B binding site and the CYBB promoter region. Inhibition of NF-B activity by salicylate reduced CYBB expression in peripheral blood neutrophils and differentiated U937 monocytic leukemia cells. U937 cells transfected with a mutant inhibitor of B super-repressor showed markedly diminished CYBB expression. Luciferase reporter analysis of the NF-B site linked to the CYBB 5 flanking promoter region revealed enhanced expression, augmented by treatment with interferon-. These studies indicate a role for this distant, 15kb upstream, binding site in NF-B regulation of the CYBB gene, an essential component of phagocyte-mediated host defense. J. Cell. Biochem. 116: 2008-2017, 2015. (c) 2015 Wiley Periodicals, Inc. (AU)

• Processo FAPESP: 06/52483-1 - Desenvolvimento e regulacao do sistema nadph oxidase humano: 1) o papel do fator nuclear kappa b (nf-kb). 2) o efeito do bay 41-2272 e doadores de oxido nitrico.
• Beneficiário: Antonio Condino Neto
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 01/14365-3 - Defeitos genético-moleculares: estudo bioquímico e características clínicas de pacientes com doença granulomatosa crônica no Brasil
• Beneficiário: Antonio Condino Neto
• Modalidade de apoio: Auxílio à Pesquisa - Regular