Miura M, Tamura T, Mikoshiba K
Department of Biological Regulation, National Institute for Basic Biology, Okazaki, Japan.
J Neurochem. 1990 Oct;55(4):1180-8. doi: 10.1111/j.1471-4159.1990.tb03123.x.
We have determined the whole promoter sequence and the transcriptional startpoint of the mouse glial fibrillary acidic protein (GFAP) gene and characterized the promoter function. We found that the cis elements for astrocyte specific expression are located within 256 bp from the transcription startpoint. We defined by DNase I footprinting assay three trans-acting factor binding sites (GFI, GFII, and GFIII) using brain or C6 astrocytoma nuclear extracts. GFI, GFII, and GFIII have AP-2, NFI, and cyclic AMP-responsive element motifs, respectively. Mutations in GFII drastically decreased the promoter activity. Base substitution in GFI and GFIII abolished the cell-specific expression, resulting in the GFAP promoter expression even in some non-GFAP-producing cells.
我们已经确定了小鼠胶质纤维酸性蛋白(GFAP)基因的完整启动子序列和转录起始点,并对启动子功能进行了表征。我们发现,星形胶质细胞特异性表达的顺式元件位于转录起始点上游256 bp范围内。我们通过DNA酶I足迹分析,使用脑或C6星形胶质细胞瘤核提取物定义了三个反式作用因子结合位点(GFI、GFII和GFIII)。GFI、GFII和GFIII分别具有AP-2、NFI和环磷酸腺苷反应元件基序。GFII中的突变显著降低了启动子活性。GFI和GFIII中的碱基替换消除了细胞特异性表达,导致即使在一些不产生GFAP的细胞中也有GFAP启动子表达。