Harrison S D, Travers A A
MRC Laboratory of Molecular Biology, Cambridge, UK.
Nucleic Acids Res. 1988 Dec 23;16(24):11403-16. doi: 10.1093/nar/16.24.11403.
With a view to identifying proteins that regulate the expression of the Drosophila ftz gene we have sequenced its enhancer-like upstream element (USE) and determined the binding sites for embryonic nuclear proteins within this region by in vitro DNAaseI footprinting. We find that greater than 50% of this element is bound by nuclear protein. By footprinting and gel-retardation studies in embryonic extracts from different developmental stages, we have characterised a number of USE/protein complexes whose nature alters in concert with changes in the ftz expression pattern, suggesting that these USE-binding proteins may be involved in the regulation of gene activity. In some cases this suggestion is substantiated by the observation that the protected DNA sequences show homology to the binding sites for ftz regulating DNA-binding proteins such as the pair-rule gene product even-skipped.
为了鉴定调控果蝇ftz基因表达的蛋白质,我们对其增强子样上游元件(USE)进行了测序,并通过体外DNA酶I足迹法确定了该区域内胚胎核蛋白的结合位点。我们发现该元件超过50%被核蛋白结合。通过对不同发育阶段胚胎提取物进行足迹法和凝胶阻滞研究,我们鉴定了一些USE/蛋白质复合物,其性质随着ftz表达模式的变化而协同改变,这表明这些USE结合蛋白可能参与了基因活性的调控。在某些情况下,这一推测得到了如下观察结果的证实:受保护的DNA序列与ftz调控DNA结合蛋白(如成对规则基因产物even-skipped)的结合位点具有同源性。