Bray S J, Johnson W A, Hirsh J, Heberlein U, Tjian R
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
EMBO J. 1988 Jan;7(1):177-88. doi: 10.1002/j.1460-2075.1988.tb02798.x.
A cis-acting sequence from the Drosophila melanogaster dopa decarboxylase (Ddc) gene is selectively required for Ddc expression in the central nervous system. We analyze several parameters influencing the function of the sequence element and describe a factor which interacts with it and mediates CNS expression of Ddc. The element, element I, can function in vivo when included on a synthetic oligonucleotide inserted near its normal location, or closer to the RNA startpoint. It displays partial activity when inverted. Two different 2-bp mutations in element I abolish its ability to stimulate neuronal Ddc expression in the CNS. A factor present in embryonic nuclear extracts specifically protects element I in DNase I footprinting assays. The binding affinity of this factor is reduced by each alteration of element I that inhibits neuronal expression, indicating a role in mediating CNS expression of Ddc. Element I alone has no detectable activity when placed adjacent to a heterologous promoter, although 2.2 kb of 5' Ddc sequences direct correct cell-specific expression of a heterologous promoter.
来自黑腹果蝇多巴脱羧酶(Ddc)基因的一个顺式作用序列是中枢神经系统中Ddc表达所特需的。我们分析了影响该序列元件功能的几个参数,并描述了一个与之相互作用并介导Ddc中枢神经系统表达的因子。当元件I包含在插入其正常位置附近或更靠近RNA起始点的合成寡核苷酸上时,它能在体内发挥作用。当它反向时表现出部分活性。元件I中的两种不同的2碱基对突变消除了其在中枢神经系统中刺激神经元Ddc表达的能力。在胚胎核提取物中存在的一种因子在DNase I足迹分析中能特异性地保护元件I。元件I的每次抑制神经元表达的改变都会降低该因子的结合亲和力,表明其在介导Ddc中枢神经系统表达中起作用。当元件I单独置于异源启动子附近时没有可检测到的活性,尽管2.2 kb的5' Ddc序列可指导异源启动子正确的细胞特异性表达。