Tolias P P, Kafatos F C
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
EMBO J. 1990 May;9(5):1457-64. doi: 10.1002/j.1460-2075.1990.tb08262.x.
We have fused various DNA sequences located upstream of the Drosophila melanogaster s36 chorion gene TATA box to a heterologous basal promoter and reporter gene (hsp70/lacZ). The expression of these constructs, following P-element-mediated germline transformation, was examined in 144 independent lines by histological staining of dissected ovaries for beta-galactosidase activity. A short 84 bp segment of the proximal 5' flanking DNA was sufficient to confer a wild-type gene expression pattern, including temporal specificity for early choriogenic follicles. Surprisingly, initial expression was very localized at the anterior and posterior poles of the follicle. The downstream half of that DNA segment permitted expression at both poles, but especially at the anterior tip, while the upstream half only favored expression in the posterior pole; these results suggested the existence of multiple, spatially specific cis-regulatory elements. When the proximal 84 bp segment was placed 1.5 kb upstream of the basal promoter, beta-galactosidase activity was observed in an altered spatial pattern, indicating that the cis-regulatory element(s) that favor expression in the posterior half of the follicle are position independent, while the element(s) that favor expression elsewhere in the follicle are position sensitive. A distal regulatory segment containing redundant DNA element(s) specific for expression in the anterior pole was identified much further upstream of s36. Thus, the expression of this chorion gene throughout the follicular epithelium is actually composite, occurring in distinct spatial domains under the control of corresponding DNA elements.
我们将位于黑腹果蝇s36绒毛膜基因TATA框上游的各种DNA序列与一个异源基础启动子和报告基因(hsp70/lacZ)融合。通过对解剖后的卵巢进行β-半乳糖苷酶活性的组织学染色,在144个独立品系中检测了这些构建体在P因子介导的种系转化后的表达情况。近端5'侧翼DNA的一个短的84 bp片段足以赋予野生型基因表达模式,包括对早期成膜卵泡的时间特异性。令人惊讶的是,初始表达非常局限于卵泡的前后极。该DNA片段的下游一半允许在两极表达,但特别是在前尖端,而上游一半仅有利于在后极表达;这些结果表明存在多个空间特异性顺式调控元件。当近端84 bp片段置于基础启动子上游1.5 kb处时,观察到β-半乳糖苷酶活性呈现出改变的空间模式,这表明有利于在卵泡后半部表达的顺式调控元件与位置无关,而有利于在卵泡其他部位表达的元件对位置敏感。在s36上游更远的位置鉴定出一个远端调控片段,其含有对在前极表达具有特异性的冗余DNA元件。因此,这个绒毛膜基因在整个卵泡上皮中的表达实际上是复合的,在相应DNA元件的控制下在不同的空间结构域中发生。