Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-κB Binding Site.
Regulation of CYBB Gene Expression in Human Phagocytes by a Distant Upstream NF-κB Binding Site.
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DOI：
10.1002/jcb.25155
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发表时间：
2015-09
影响因子：
4
通讯作者：
Newburger PE
中科院分区：
文献类型：
--
作者：
Frazão JB;Thain A;Zhu Z;Luengo M;Condino-Neto A;Newburger PE
人CYBB基因编码吞噬细胞氧化酶复合物的gp 91-phox组分，其负责产生超氧化物和其他对微生物杀灭至关重要的下游活性氧物质。在本研究中，我们通过序列分析确定了位于CYBB基因远端5′侧翼区的DNase I超敏位点（称为HS-II）中的一个推定的NF-κB结合位点。电泳迁移率分析显示重组NF-κB蛋白p50和HL-60髓性白血病细胞系核提取物中对应于p50和p50/p65异二聚体的蛋白质与序列元件结合。染色质免疫沉淀证实NF-κB与完整HL-60细胞中的该位点结合。染色体构象捕获（3C）分析证明NF-κB结合位点和CYBB启动子区域之间存在物理相互作用。水杨酸抑制NF-κB活性可降低外周血中性粒细胞和分化的U937单核细胞白血病细胞中CYBB的表达。用κB“超级阻遏物”的突变体抑制剂转染的U937细胞显示CYBB表达显著降低。荧光素酶报告基因分析显示，与CYBB 5′侧翼启动子区连接的NF-κB位点表达增强，干扰素-γ处理后表达增强。这些研究表明，这个遥远的上游15 kb结合位点在CYBB基因的NF-κB调节中发挥作用，CYBB基因是吞噬细胞介导的宿主防御的重要组成部分。
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-κB 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-κB 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-κB binding to the site in intact HL-60 cells. Chromosome conformation capture (3C) assays demonstrated physical interaction between the NF-κB binding site and the CYBB promoter region. Inhibition of NF-κB activity by salicylate reduced CYBB expression in peripheral blood neutrophils and differentiated U937 monocytic leukemia cells. U937 cells transfected with a mutant inhibitor of κB “super-repressor” showed markedly diminished CYBB expression. Luciferase reporter analysis of the NF-κB site linked to the CYBB 5′ flanking promoter region revealed enhanced expression, augmented by treatment with interferon-γ. These studies indicate a role for this distant, 15 kb upstream, binding site in NF-κB regulation of the CYBB gene, an essential component of phagocyte-mediated host defense.
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影响因子：
7.3
作者：
Bhatt D;Ghosh S
通讯作者：
Ghosh S
影响因子：
5.2
作者：
Bustamante J;Picard C;Boisson-Dupuis S;Abel L;Casanova JL
通讯作者：
Casanova JL
影响因子：
4.8
作者：
Hawkins, SM;Kohwi-Shigematsu, T;Skalnik, DG
通讯作者：
Skalnik, DG
影响因子：
4.8
作者：
Anrather, J;Racchumi, G;Iadecola, C
通讯作者：
Iadecola, C
DOI：
10.1073/pnas.85.14.5215
发表时间：
1988-07-01
影响因子：
11.1
作者：
NEWBURGER, PE;EZEKOWITZ, RAB;ORKIN, SH
通讯作者：
ORKIN, SH