Aguanno A, Lee M R, Marden C M, Rattray M, Gault A, Albert V R
Roche Institute of Molecular Biology, Nutley, NJ 07110, USA.
J Neurochem. 1995 Nov;65(5):1944-54. doi: 10.1046/j.1471-4159.1995.65051944.x.
The rat aromatic L-amino acid decarboxylase (AADC) gene contains alternative promoters directing expression of neuronal and nonneuronal mRNAs that differ only in their 5' untranslated regions (UTRs). We have analyzed the expression of the neuronal promoter of the AADC gene in cells synthesizing catecholamines and serotonin, as well as in non-AADC-expressing cells. We demonstrate the use of the neuronal-specific UTR in individual dopamine-, norepinephrine-, and serotonin-containing neurons. Transfection analyses show that the rat AADC neuronal promoter, containing 2,400 bp upstream of the transcription start site and including the 68-bp untranslated exon 2, can activate transcription from a reporter gene in both catecholaminergic and serotonergic cell lines. These analyses identified several positive and negative cis-active elements within this region. Unexpectedly, we observed that this promoter, when removed from its native context within the AADC gene, can also direct expression of a reporter gene in cells that do not normally express AADC mRNA. These results suggest that tissue-specific expression of the neuronal promoter may not be controlled by cis-active elements within the first 2,400 bp of the promoter. Additional information may be required to restrict neuronal promoter expression to appropriate cell types. This regulatory information could reside elsewhere within the promoter, within introns, or may be provided by interactions between the two AADC promoters.
大鼠芳香族L-氨基酸脱羧酶(AADC)基因含有不同的启动子,可指导仅在5'非翻译区(UTR)存在差异的神经元和非神经元mRNA的表达。我们分析了AADC基因的神经元启动子在合成儿茶酚胺和5-羟色胺的细胞以及不表达AADC的细胞中的表达情况。我们证明了在单个含多巴胺、去甲肾上腺素和5-羟色胺的神经元中使用神经元特异性UTR。转染分析表明,大鼠AADC神经元启动子在转录起始位点上游含有2400 bp,包括68 bp的非翻译外显子2,可在儿茶酚胺能和5-羟色胺能细胞系中激活报告基因的转录。这些分析确定了该区域内的几个正向和负向顺式作用元件。出乎意料的是,我们观察到该启动子从其在AADC基因中的天然背景中移除后,也可在通常不表达AADC mRNA的细胞中指导报告基因的表达。这些结果表明,神经元启动子的组织特异性表达可能不受启动子前2400 bp内顺式作用元件的控制。可能需要额外的信息将神经元启动子的表达限制在适当的细胞类型中。这种调控信息可能存在于启动子内的其他位置、内含子内,或者可能由两个AADC启动子之间的相互作用提供。