Johnson W A, McCormick C A, Bray S J, Hirsh J
Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Boston, Massachusetts 02115.
Genes Dev. 1989 May;3(5):676-86. doi: 10.1101/gad.3.5.676.
At least two cis-regulatory elements are necessary for correct neuron-specific expression of the Drosophila melanogaster dopa decarboxylase gene, Ddc. In addition to a previously described proximal element located approximately 60 bp upstream of the mRNA start site, we have now characterized a distal approximately 600-bp DNA fragment, extending from -1019 to -1623 bp, which possesses enhancer-like properties and is essential for normal neuron-specific expression. Immunofluorescent labeling of neurons expressing deleted Ddc genes indicates that this region contains both general neuronal regulatory elements and cell-specific elements that selectively affect Ddc expression in either dopaminergic or serotonergic neurons. These selective effects can be correlated with the removal of sequence elements that are protected from DNase digestion by factors present in embryonic nuclear extracts. Several of these elements are also homologous to sequences located upstream of the evolutionarily diverged Ddc gene of Drosophila virilis. These results suggest that the neuron-specific expression of Ddc results from the combined action of several factors binding within this distal enhancer region.
至少两个顺式调控元件对于果蝇多巴脱羧酶基因(Ddc)正确的神经元特异性表达是必需的。除了先前描述的位于mRNA起始位点上游约60 bp处的近端元件外,我们现在还鉴定了一个约600 bp的远端DNA片段,其范围从-1019至-1623 bp,该片段具有增强子样特性,并且对于正常的神经元特异性表达至关重要。对表达缺失Ddc基因的神经元进行免疫荧光标记表明,该区域既包含一般的神经元调控元件,也包含在多巴胺能或5-羟色胺能神经元中选择性影响Ddc表达的细胞特异性元件。这些选择性作用可能与某些序列元件的去除有关,这些序列元件在胚胎核提取物中存在的因子作用下可免受DNA酶消化。其中几个元件也与果蝇Ddc基因进化上分化的上游序列同源。这些结果表明,Ddc的神经元特异性表达是由结合在该远端增强子区域内的几种因子共同作用的结果。