Analysis of promoter regions in Hailey-Hailey disease and Darier's disease genes.

2003年的研究结果：1）在Hailey-Hailey disease（HHD）基因ATP 2C 1启动子区（从非编码区到-1200bp）发现了6个SNPs。2）将ATP 2C 1基因启动子区的不同片段克隆到荧光素酶载体中，利用HaCat细胞进行报告基因分析。我们已经确定+22至+60片段作为ATP 2C 1的表达控制元件。利用抗体和突变体诱饵DNA片段的另外的凝胶筛选测定揭示了两种转录因子，SP-1和YY-1，2004年的研究结果表明：1）将SP-1或YY-1克隆与ATP 2C 1启动子共转染入细胞，可刺激ATP 2C 1的表达; 3）SP-1在HHD角质形成细胞中的表达较低，并且由于其异常稳定的蛋白质表达而对培养物中升高的细胞外钙浓度没有反应。ATP 2C 1基因突变导致细胞胞浆内钙离子浓度升高。

Results obtained by research in 2003.1) We have identified 6 SNPs on the promoter region (from non-cording to -1200bp) of ATP2C1 (Hailey-Hailey disease (HHD) gene). There was no correlation between these SNPs and disease phenotypes of HHD.2) Various segments of promoter region of ATP2C1 were cloned into Luciferase vector, and reporter assay was performed using HaCat cell. We have identified +22 to +60 by segment as a expression controlling element for ATP2C1. Additional gel-sift assay utilizing antibodies and mutant decoy DNA segments revealed that two transcription factors, SP-1 and YY-1, could bind to this promoter region.Results obtained by research in 2004.1) Co-transfection of ATP2C1 promoter and SP-1 or YY-1 clone into the cells stimulated the expression of ATP2C1.2) Expression of SP-1 itself was up regulated by raised extracellular calcium concentration in the culture of HaCat cells.3) Expression of SP-1 was low in HHD keratinocytes and did not respond to raised extracellular calcium concentration in the culture due to abnormally stable-high level of calcium concentration in the cytosole of the cells caused by genetic mutation of ATP2C1.