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

2003.1)我们在Hailey-Hailey病(HHD)基因启动子区(非编码区至-1200bp)发现了6个SNPs。2)将ATP2C1启动子区不同片段克隆到荧光素酶载体上，用HaCat细胞进行报告基因分析。我们已经通过片段确定+22到+60是ATP2C1的表达控制元件。另外，利用抗体和突变诱骗DNA片段进行的凝胶筛选试验显示，两个转录因子SP-1和YY-1，结果：2004.1)将ATP2C1启动子与SP-1或YY-1克隆共转入细胞内，可刺激ATP2C1mRNA的表达。2)在HaCat细胞培养中，SP-1的表达随细胞外钙浓度的升高而上调。3)角质形成细胞中SP-1的表达水平较低，对细胞外钙离子浓度的升高无反应，这是由于ATP2C1基因突变导致细胞胞浆内钙浓度异常稳定-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.