Title
Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-kappaB Binding Site
UMMS Affiliation
Department of Pediatrics, Division of Hematology/Oncology; Department of Molecular, Cellular, and Cancer Biology
Publication Date
2015-09-01
Document Type
Article
Disciplines
Biochemistry | Cell Biology
Abstract
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-kappaB 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-kappaB 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-kappaB binding to the site in intact HL-60 cells. Chromosome conformation capture (3C) assays demonstrated physical interaction between the NF-kappaB binding site and the CYBB promoter region. Inhibition of NF-kappaB activity by salicylate reduced CYBB expression in peripheral blood neutrophils and differentiated U937 monocytic leukemia cells. U937 cells transfected with a mutant inhibitor of kappaB "super-repressor" showed markedly diminished CYBB expression. Luciferase reporter analysis of the NF-kappaB site linked to the CYBB 5' flanking promoter region revealed enhanced expression, augmented by treatment with interferon-gamma. These studies indicate a role for this distant, 15 kb upstream, binding site in NF-kappaB regulation of the CYBB gene, an essential component of phagocyte-mediated host defense.
DOI of Published Version
10.1002/jcb.25155
Source
J Cell Biochem. 2015 Sep;116(9):2008-17. doi: 10.1002/jcb.25155. Link to article on publisher's site
Journal/Book/Conference Title
Journal of cellular biochemistry
Related Resources
PubMed ID
25752509
Repository Citation
Frazao, Josias B.; Thain, Alison; Zhu, Zhiqing; Luengo, Marcos; Condino-Neto, Antonio; and Newburger, Peter E., "Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-kappaB Binding Site" (2015). Pediatric Publications and Presentations. 49.
https://escholarship.umassmed.edu/peds_pp/49