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dc.contributor.authorFrazao, Josias B.
dc.contributor.authorThain, Alison
dc.contributor.authorZhu, Zhiqing
dc.contributor.authorLuengo, Marcos
dc.contributor.authorCondino-Neto, Antonio
dc.contributor.authorNewburger, Peter E.
dc.date2022-08-11T08:10:13.000
dc.date.accessioned2022-08-23T17:00:09Z
dc.date.available2022-08-23T17:00:09Z
dc.date.issued2015-09-01
dc.date.submitted2016-11-07
dc.identifier.citationJ Cell Biochem. 2015 Sep;116(9):2008-17. doi: 10.1002/jcb.25155. <a href="http://dx.doi.org/10.1002/jcb.25155">Link to article on publisher's site</a>
dc.identifier.issn0730-2312 (Linking)
dc.identifier.doi10.1002/jcb.25155
dc.identifier.pmid25752509
dc.identifier.urihttp://hdl.handle.net/20.500.14038/43735
dc.description.abstractThe 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.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=25752509&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1002/jcb.25155
dc.subjectBiochemistry
dc.subjectCell Biology
dc.titleRegulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-kappaB Binding Site
dc.typeArticle
dc.source.journaltitleJournal of cellular biochemistry
dc.source.volume116
dc.source.issue9
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/peds_pp/49
dc.identifier.contextkey9353390
html.description.abstract<p>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.</p>
dc.identifier.submissionpathpeds_pp/49
dc.contributor.departmentDepartment of Molecular, Cellular, and Cancer Biology
dc.contributor.departmentDepartment of Pediatrics, Division of Hematology/Oncology
dc.source.pages2008-17


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