Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene
Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene
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DOI：
10.1152/physiolgenomics.00005.2011
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发表时间：
2011-07-01
影响因子：
4.6
通讯作者：
Umemura, Satoshi
中科院分区：
文献类型：
--
作者：
Matsuda, Miyuki;Tamura, Kouichi;Umemura, Satoshi
Matsuda M，Tamura K，Wakui H，Dejima T，Maeda A，Ohsawa M，Kanaoka T，Haku S，Azushima K，Yamasaki H，Saito D，Hirose T，Maeshima Y，Nagashima Y，Umemura S. Runx3参与小鼠血管紧张素II 1型受体相关蛋白基因的基础转录激活。 Physiol Genomics 43: 884-894, 2011。首次发表于 2011 年 5 月 17 日； doi:10.1152/physicalgenomics.00005.2011.-我们之前克隆了一种与血管紧张素 II 1 型 (AT1) 受体相互作用的分子，对 AT1 受体信号传导发挥抑制功能，我们将其命名为 ATRAP/Agtrap（AT1 受体相关蛋白）。在本研究中，我们使用肾远曲小管细胞检查了基础 ATRAP 基因表达的调节。我们发现血清饥饿上调了 ATRAP 基因的基础表达，这是一种需要从头合成 mRNA 和蛋白质的反应。荧光素酶测定表明，近端启动子区域指导转录，并且 runt 相关转录因子 (RBE) 的推定结合位点对于转录激活很重要。 RBE-decoy 转染和小干扰 RNA 内源性敲低的结果表明，runt 相关转录因子 Runx3 参与 ATRAP 基因表达。染色质免疫沉淀测定也支持 Runx3 与肾远曲小管细胞中 ATRAP 启动子的结合。免疫组织化学显示连续切片中肾远端曲管和连接小管中 Runx3 和 ATRAP 蛋白的表达。此外，单侧输尿管梗阻 7 天后，受影响肾脏中 Runx3 免疫染色降低，同时 ATRAP 表达受到抑制。这些发现表明 Runx3 在体外和体内肾远端肾小管细胞的 ATRAP 基因表达中发挥作用。
Matsuda M, Tamura K, Wakui H, Dejima T, Maeda A, Ohsawa M, Kanaoka T, Haku S, Azushima K, Yamasaki H, Saito D, Hirose T, Maeshima Y, Nagashima Y, Umemura S. Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene. Physiol Genomics 43: 884-894, 2011. First published May 17, 2011; doi:10.1152/physiolgenomics.00005.2011.-We previously cloned a molecule that interacts with angiotensin II type 1 (AT1) receptor to exert an inhibitory function on AT1 receptor signaling that we named ATRAP/Agtrap (for AT1 receptor-associated protein). In the present study we examined the regulation of basal ATRAP gene expression using renal distal convoluted tubule cells. We found that serum starvation upregulated basal expression of ATRAP gene, a response that required de novo mRNA and protein synthesis. Luciferase assay revealed that the proximal promoter region directs transcription and that a putative binding site of runt-related transcription factors (RBE) is important for transcriptional activation. The results of RBE-decoy transfection and endogenous knockdown by small interference RNA showed that the runt-related transcription factor Runx3 is involved in ATRAP gene expression. Chromatin immunoprecipitation assay also supported the binding of Runx3 to the ATRAP promoter in renal distal convoluted tubule cells. Immunohistochemistry demonstrated the expression of Runx3 and ATRAP proteins in the distal convoluted and connecting tubules of the kidney in consecutive sections. Furthermore, the Runx3 immunostaining was decreased together with a concomitant suppression of ATRAP expression in the affected kidney after 7 days of unilateral ureteral obstruction. These findings indicate that Runx3 plays a role in ATRAP gene expression in renal distal tubular cells both in vitro and in vivo.