Transcriptional control of receptor activator of nuclear factor-κB ligand by the protein kinase A activator forskolin and the transmembrane glycoprotein 130-activating cytokine, oncostatin M, is exerted through multiple distal enhancers
Transcriptional control of receptor activator of nuclear factor-κB ligand by the protein kinase A activator forskolin and the transmembrane glycoprotein 130-activating cytokine, oncostatin M, is exerted through multiple distal enhancers
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DOI：
10.1210/me.2006-0315
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发表时间：
2007-01-01
影响因子：
--
通讯作者：
Pike, J. Wesley
中科院分区：
文献类型：
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作者：
Kim, Sungtae;Yamazaki, Miwa;Pike, J. Wesley
核因子-kappa B受体激活剂配体(RANKL)是一种强有力的破骨细胞因子，其表达受1，25-二羟基维生素D-3[1，25-(OH)(2)D-3]、蛋白激酶A(PKA)激活剂如甲状旁腺素(PTH)和跨膜糖蛋白130(Gp130)激活的细胞因子如oncoatin M等转录水平的调节。我们最近在RANKL转录起始点上游发现了5个高度保守的染色质结构域，它们有助于1，25-(OH)(2)D-3及其受体增强RANKL基因的输出。因此，我们使用针对PKA靶标cAMP反应元件结合蛋白(CREB)和gp130靶标信号转导和转录激活子3的抗体的定向染色质免疫沉淀方法筛选了这五个共同的调节区，以了解它们介导PKA和gp130激活子作用的潜在能力。CREB在每个上游调节区都被鉴定出来；相反，信号转导和转录激活子3只与一个亚集相关。有趣的是，在异源转染系统中，只有这些区域的最远端显示出CREB和抑癌素M调节的转录活性。图谱研究指出，两个高度保守的cAMP反应元件以及一个相邻的调控位点结合了Run转录因子2，并能够影响基础和激素诱导的RANKL活性。令人惊讶的是，PKA和gp130的激活促进了RNA聚合酶II在五个末端增强子以及RANKL转录起始点的招募。激活还伴随着组蛋白4乙酰化的显著和位置选择性升高。这项研究表明，PKA和gp130诱导的RANKL基因表达是通过共同的调控结构域介导的，这些结构域也有助于1，25-(OH)(2)D-3的活性。
Receptor activator of nuclear factor-kappa B ligand (RankL) is a potent osteoclastogenic cytokine the expression of which is regulated at the transcriptional level by 1,25-dihydroxyvitamin D-3 [1,25-(OH)(2)D-3], protein kinase A (PKA) activators such as PTH and transmembrane glycoprotein 130 (gp130)-activating cytokines such as oncostatin M. We recently identified five highly conserved chromatin domains located significant distances upstream of the RankL transcriptional start site that contribute to the ability of 1,25-(OH)(2)D-3 and its receptor to enhance RankL gene output. We therefore screened these five common regulatory regions for their potential ability to mediate the actions of PKA- and gp130-activators using a directed chromatin immunoprecipitation approach employing antibodies to the PKA target cAMP response element-binding protein (CREB) and the gp130 target signal transducer and activator of transcription 3. CREB was identified at each of the upstream regulatory regions; signal transducer and activator of transcription 3, in contrast, was associated with only a subset. Interestingly, only the most distal of these regions demonstrated CREB-and oncostatin M-regulated transcriptional activity in a heterologous transfection system. Mapping studies pointed to two highly conserved cAMP response elements as well as an adjacent regulatory site that bound Runt transcription factor 2 and was able to influence both basal as well as hormone-inducible RankL activity. Surprisingly, PKA and gp130 activation prompted recruitment of RNA polymerase II to the five distal enhancers as well as to the RankL transcriptional start site. Activation was also accompanied by a significant and location-selective rise in histone 4 acetylation. This study demonstrates that the activation of RankL gene expression by PKA- and gp130-inducers is mediated via common regulatory domains that also served to facilitate the activity of 1,25-(OH)(2)D-3.