Histone acetylation and activation of cAMP-response element-binding protein regulate transcriptional activation of MKP-M in lipopolysaccharide-stimulated macrophages
Histone acetylation and activation of cAMP-response element-binding protein regulate transcriptional activation of MKP-M in lipopolysaccharide-stimulated macrophages
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DOI：
10.1074/jbc.m211829200
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发表时间：
2003-03-14
影响因子：
4.8
通讯作者：
Matsuguchi, T
中科院分区：
文献类型：
--
作者：
Musikacharoen, T;Yoshikai, Y;Matsuguchi, T
MRP-M 是一种双特异性磷酸酶，可优先灭活 JNK。 mkp-M 基因表达由巨噬细胞中的脂多糖 (LPS) 刺激快速诱导，并参与 LPS 介导的 JNK 激活和肿瘤坏死因子 -a 分泌的负调节。为了揭示 mkp-M 基因的转录调控，我们分离了小鼠 mkp-M 基因并绘制了其转录起始位点。将含有 mkp-M 基因 5'-上游区域的荧光素酶报告质粒稳定转染至 RAW264.7 细胞中。使用这些细胞的测定表明，mkp-M 启动子激活需要 -252 和 -135 之间的启动子区域。测序分析揭示了该区域的 E box 和 CREB 响应元件，而电动迁移试验和诱变证实这两个元件对于 mkp-M 基因的 LPS 响应性至关重要。我们还利用染色质免疫沉淀分析发现，LPS 刺激导致 RAW264.7 细胞中 mkp-M 启动子处的组蛋白 H3 和 114 乙酰化。与此一致的是，组蛋白脱乙酰酶抑制剂曲古抑菌素 A 增加了内源性 mkp-M 基因的转录。最后，DNase I 超敏位点定位揭示了 E 盒和 CREB 反应元件周围 LPS 刺激后可诱导的超敏位点。总之，我们的数据表明 LPS 对巨噬细胞中 mkp-M 基因转录的激活与组蛋白乙酰化和染色质重塑有关。
MRP-M is a dual specificity phosphatase that preferentially inactivates JNK. mkp-M gene expression is rapidly induced by lipopolysaccharide (LPS) stimulation in macrophages and is involved in the negative regulation of LPS-mediated JNK activation and tumor necrosis factor-a secretion. To reveal the transcriptional regulation of the mkp-M gene, we isolated the mouse mkp-M gene and mapped its transcriptional start site. Luciferase reporter plasmids containing 5'-upstream regions of the mkp-M gene were stably transfected into RAW264.7 cells. The assays using these cells revealed that the promoter region between -252 and -135 is required for mkp-M promoter activation. Sequencing analysis revealed E box and CREB-responsive elements in this region, and electromobility shift assays and mutagenesis confirmed that both of these elements are essential for LPS responsiveness of the mkp-M gene. We also utilized chromatin immunoprecipitation assay and found that LPS stimulation caused acetylation of histone H3 and 114 at mkp-M promoter in RAW264.7 cells. Consistent with this, a histone deacetylase inhibitor, trichostatin A, increased endogenous mkp-M gene transcription. Finally, DNase I hypersensitivity site mapping revealed the inducible hypersensitivity site after LPS stimulation around the location of the E box and CREB-responsive elements. Altogether, our data indicated that the activation of mkp-M gene transcription in macrophages by LPS is associated with histone acetylation and chromatin remodeling.