McKenzie R W, Hu J, Brennan M D
Department of Biochemistry, School of Medicine, University of Louisville, KY 40292.
Nucleic Acids Res. 1994 Apr 11;22(7):1257-64. doi: 10.1093/nar/22.7.1257.
The alcohol dehydrogenase (Adh) gene in the Hawaiian species of fruit fly, Drosophila affinidisjuncta, like the Adh genes from all Drosophila species analyzed, is expressed at high levels in the larval fat body via a larval-specific promoter. To identify the cis-acting elements involved in this highly conserved aspect of Adh gene expression, deleted D. affinidisjuncta genes were introduced into D. melanogaster by somatic transformation. Unlike previously described methods, this transformation system allows analysis of Adh gene expression specifically in the larval fat body. The arrangement of sequences influencing expression of the proximal promoter of this gene in the larval fat body differs markedly from that described for the Adh gene from the distant relative, D. melanogaster. Multiple redundant elements dispersed 5' and 3' to the gene, only some of which map to regions carrying evolutionarily conserved sequences, affect expression in the fat body. D. affinidisjuncta employs a novel mode of Adh gene regulation in which the proximal promoter is influenced by sequences having roles in expression of the distal promoter. This gene is also unique in that far upstream sequences can compensate for loss of sequences within 200 bp of the proximal RNA start site. Furthermore, expression is influenced in an unusual, context-dependent manner by a naturally-occurring 3' duplication of the proximal promoter--a feature found only in Hawaiian species.
夏威夷果蝇物种——间断亲和果蝇(Drosophila affinidisjuncta)中的乙醇脱氢酶(Adh)基因,与所有已分析的果蝇物种的Adh基因一样,通过幼虫特异性启动子在幼虫脂肪体中高水平表达。为了鉴定参与Adh基因表达这一高度保守方面的顺式作用元件,通过体细胞转化将缺失的间断亲和果蝇基因导入黑腹果蝇(D. melanogaster)。与先前描述的方法不同,该转化系统允许专门分析幼虫脂肪体中的Adh基因表达。影响该基因近端启动子在幼虫脂肪体中表达的序列排列,与远亲黑腹果蝇的Adh基因所描述的排列明显不同。多个冗余元件分散在基因的5'和3'端,其中只有一些定位于携带进化保守序列的区域,影响脂肪体中的表达。间断亲和果蝇采用了一种新的Adh基因调控模式,其中近端启动子受在远端启动子表达中起作用的序列影响。该基因的独特之处还在于,远上游序列可以补偿近端RNA起始位点200 bp内序列的缺失。此外,近端启动子自然发生的3'重复以一种不寻常的、依赖上下文的方式影响表达——这一特征仅在夏威夷物种中发现。