Combined ChIP-Seq and transcriptome analysis identifies AP-1/JunD as a primary regulator of oxidative stress and IL-1β synthesis in macrophages.
Combined ChIP-Seq and transcriptome analysis identifies AP-1/JunD as a primary regulator of oxidative stress and IL-1β synthesis in macrophages.
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DOI：
10.1186/1471-2164-14-92
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发表时间：
2013-02-11
期刊：
影响因子：
4.4
通讯作者：
Behmoaras J
中科院分区：
文献类型：
--
作者：
Hull RP;Srivastava PK;D'Souza Z;Atanur SS;Mechta-Grigoriou F;Game L;Petretto E;Cook HT;Aitman TJ;Behmoaras J
氧化爆发是巨噬细胞采用的主要抗菌机制之一。 WKY 大鼠品系对实验诱导的巨噬细胞依赖性新月体肾小球肾炎 (Crgn) 特别敏感。我们之前发现 AP-1 转录因子 JunD 是 WKY 骨髓源性巨噬细胞 (BMDM) 中巨噬细胞活化的决定因素。 JunD 在 WKY BMDM 中过度表达，其沉默会减少这些细胞中 Fc 受体介导的氧化爆发。在这里，我们将 Jund RNA 干扰与微阵列分析以及 WKY BMDM 中的 ChIP 测序 (ChIP-Seq) 分析结合起来，以研究 JunD 介导的基础细胞和脂多糖 (LPS) 刺激细胞中巨噬细胞活化的控制。 Jund 沉默后的微阵列分析表明，Jund 激活和抑制基因表达，与氧化应激和 IL-1β 表达相关的基因具有显着差异表达（>3 倍）。通过将 WKY BMDM 中的全基因组表达与表达较低水平 JunD 的 Jund 同系株 (WKY.LCrgn2) BMDM 进行比较来补充这些结果。 ChIP-Seq 分析表明，与 WKY.LCrgn2 BMDM 相比，JunD 表达增加导致 WKY BMDM 中结合事件数量增加。结合 ChIP-Seq 和微阵列分析揭示了一组主要的 JunD 靶标，JunD 通过这些靶标对基础巨噬细胞和 LPS 刺激的巨噬细胞中的氧化应激和 IL-1β 合成发挥作用。这些发现证明了基因决定的转录因子水平如何影响其在原代细胞中的结合位点，并确定 JunD 是原代巨噬细胞中氧化应激和 IL-1β 合成的关键调节因子，这可能在 Crgn 的易感性中发挥作用。
The oxidative burst is one of the major antimicrobial mechanisms adopted by macrophages. The WKY rat strain is uniquely susceptible to experimentally induced macrophage-dependent crescentic glomerulonephritis (Crgn). We previously identified the AP-1 transcription factor JunD as a determinant of macrophage activation in WKY bone marrow-derived macrophages (BMDMs). JunD is over-expressed in WKY BMDMs and its silencing reduces Fc receptor-mediated oxidative burst in these cells. Here we combined Jund RNA interference with microarray analyses alongside ChIP-sequencing (ChIP-Seq) analyses in WKY BMDMs to investigate JunD-mediated control of macrophage activation in basal and lipopolysaccharide (LPS) stimulated cells. Microarray analysis following Jund silencing showed that Jund activates and represses gene expression with marked differential expression (>3 fold) for genes linked with oxidative stress and IL-1β expression. These results were complemented by comparing whole genome expression in WKY BMDMs with Jund congenic strain (WKY.LCrgn2) BMDMs which express lower levels of JunD. ChIP-Seq analyses demonstrated that the increased expression of JunD resulted in an increased number of binding events in WKY BMDMs compared to WKY.LCrgn2 BMDMs. Combined ChIP-Seq and microarray analysis revealed a set of primary JunD-targets through which JunD exerts its effect on oxidative stress and IL-1β synthesis in basal and LPS-stimulated macrophages. These findings demonstrate how genetically determined levels of a transcription factor affect its binding sites in primary cells and identify JunD as a key regulator of oxidative stress and IL-1β synthesis in primary macrophages, which may play a role in susceptibility to Crgn.
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DOI：
10.1152/ajplung.00315.2009
发表时间：
2010-09-01
影响因子：
4.9
作者：
Hamanaka, Kazutoshi;Jian, Ming-Yuan;Parker, James C.
通讯作者：
Parker, James C.
影响因子：
16
作者：
Heinz S;Benner C;Spann N;Bertolino E;Lin YC;Laslo P;Cheng JX;Murre C;Singh H;Glass CK
通讯作者：
Glass CK
DOI：
10.1111/j.1749-6632.2009.04813.x
发表时间：
2009-01-01
期刊：
CONTEMPORARY CHALLENGES IN AUTOIMMUNITY
影响因子：
--
作者：
Lacotte, Stephanie;Brun, Susana;Dumortier, Helene
通讯作者：
Dumortier, Helene
影响因子：
10.5
作者：
Barish, Grant D.;Yu, Ruth T.;Evans, Ronald M.
通讯作者：
Evans, Ronald M.
影响因子：
2.9
作者：
Fontayne, A;Dang, PMC;El Benna, J
通讯作者：
El Benna, J