Promoter Analysis of Mouse Scn3a Gene and Regulation of the Promoter Activity by GC Box and CpG Methylation
Promoter Analysis of Mouse Scn3a Gene and Regulation of the Promoter Activity by GC Box and CpG Methylation
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小鼠Scn3a基因启动子分析及GC Box和CpG甲基化对启动子活性的调控
DOI：
10.1007/s12031-011-9492-8
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发表时间：
2011-06-01
影响因子：
3.1
通讯作者：
Long, Yue-Sheng
中科院分区：
文献类型：
--
作者：
Deng, Guang-Fei;Qin, Jia-Ming;Long, Yue-Sheng
电压门控钠通道 α 亚基 III 型 (Na(v)1.3) 主要在中枢神经系统中表达，与神经系统疾病相关。小鼠Scn3a产物(Na(v)1.3)的表达主要发生在胚胎和出生后早期脑中，但在成年脑中不表达。在此，我们首次报道了小鼠Scn3a基因启动子区域的鉴定和表征以及GC盒和CpG甲基化对启动子活性的调节。荧光素酶检测显示，与SH-SY5Y细胞和HEK293细胞相比，PC12细胞中启动子区F1.2（nt -1,049至+157）的活性显着更高。启动子区域的逐步5'截短发现最小功能启动子位于nt -168至+157区域内。小鼠 Scn3a 启动子中 GC 盒（nt -254 至 -258）的缺失降低了启动子活性。没有 GC 盒的 F1.2 的 CpG 甲基化完全抑制了启动子活性，表明 GC 盒是 CpG 甲基化的 Scn3a 启动子中的关键元件。这些结果表明GC盒和CpG甲基化可能在调节小鼠Scn3a基因表达中发挥重要作用。
Voltage-gated sodium channel alpha-subunit type III (Na(v)1.3) is mainly expressed in the central nervous system and is associated with neurological disorders. The expression of mouse Scn3a product (Na(v)1.3) mainly occurs in embryonic and early postnatal brain but not in adult brain. Here, we report for the first time the identification and characterization of the mouse Scn3a gene promoter region and regulation of the promoter activity by GC box and CpG methylation. Luciferase assay showed that the promoter region F1.2 (nt -1,049 to +157) had significantly higher activity in PC12 cells, comparing with that in SH-SY5Y cells and HEK293 cells. A stepwise 5' truncation of the promoter region found that the minimal functional promoter located within the region nt -168 to +157. Deletion of a GC box (nt -254 to -258) in the mouse Scn3a promoter decreased the promoter activity. CpG methylation of the F1.2 without the GC box completely repressed the promoter activity, suggesting that the GC box is a critical element in the CpG-methylated Scn3a promoter. These results suggest that the GC box and CpG methylation might play important roles in regulating mouse Scn3a gene expression.