A novel E box/AT-rich element is required for muscle-specific expression of the sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene.
A novel E box/AT-rich element is required for muscle-specific expression of the sarcoplasmic reticulum Ca2+-ATPase (SERCA2) gene.
复制标题
肌浆网 Ca2-ATP 酶 (SERCA2) 基因的肌肉特异性表达需要一种新的 E 盒/富含 AT 的元件。
DOI：
10.1093/nar/26.4.1092
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发表时间：
1998
影响因子：
14.9
通讯作者：
Periasamy,M
中科院分区：
文献类型：
--
作者：
Baker,DL;Dave,V;Reed,T;Misra,S;Periasamy,M
心脏/慢肌浆网 (SR) Ca2+-ATP 酶基因 (SERCA2) 编码钙转运泵，其表达以组织和发育特异性方式进行调节。之前我们已经确定了两个不同的正调控区（bp -284 至 -72 和 -1815 至 -1105）对于 SERCA2 启动子活性很重要。在这里，我们证明SERCA2远端启动子区域通过以肌细胞特异性方式激活异源启动子（TK）来像增强子一样发挥作用。通过缺失分析，核心增强子区域被界定为-1467至-1105bp片段。我们确定位于-1115 bp 的富含 E 盒/AT 的元件对于最大增强子活性至关重要。凝胶迁移率变化研究表明，这种富含 E 盒/AT 的元件特异性结合 Sol8 肌生成过程中诱导的蛋白质。该区域包括两个其他顺式作用元件 CArG 和 MCAT，它们也结合来自 Sol8 肌管的特定核蛋白复合物。每个位点的诱变都会导致 SERCA/TK-CAT 启动子活性降低。基于这些数据，我们提出E盒/AT富含元件可能与CArG和MCAT元件一起有助于SERCA2基因启动子的整体激活和调节。
The cardiac/slow twitch sarcoplasmic reticulum (SR) Ca2+-ATPase gene (SERCA2) encodes a calcium transport pump whose expression is regulated in a tissue- and development-specific manner. Previously we have identified two distinct positive regulatory regions (bp −284 to −72 and −1815 to −1105) as important forSERCA2promoter activity. Here we demonstrate that theSERCA2distal promoter region functions like an enhancer by activating a heterologous promoter (TK) in a muscle cell-specific manner. Through deletion analysis a core enhancer region was delimited to the −1467 to −1105 bp fragment. We identified the E box/AT-rich element located at −1115 bp as critical for maximal enhancer activity. Gel mobility shift studies revealed that this E box/AT-rich element specifically binds a protein which is induced during Sol8 myogenesis. This region includes two othercis-acting elements, CArG and MCAT, which also bind specific nuclear protein complexes from Sol8 myotubes. Mutagenesis of each of these sites resulted in decreasedSERCA/TK-CATpromoter activity. Based on these data, we propose that the E box/AT-rich element may contribute along with CArG and MCAT elements to the overall activation and regulation of theSERCA2gene promoter.