Villanyi Zoltan, Papp Bernadett, Szikora Szilard, Boros Imre, Szabad Janos
Faculty of Medicine, Department of Biology, University of Szeged, Somogyi Bela str. 4, Szeged, Hungary.
Mech Dev. 2008 Sep-Oct;125(9-10):822-31. doi: 10.1016/j.mod.2008.07.001. Epub 2008 Jul 6.
Importin-beta, encoded by the Ketel gene in Drosophila, is a key component of nuclear protein import, the formation of the spindle microtubules and the assembly of the nuclear envelope. The Drosophila embryos rely on the maternal importin-beta dowry at the beginning of their life. Expression of the zygotic Ketel gene commences during gastrulation in every cell and while the expression is maintained in the mitotically active diploid cells it ceases in the non-dividing larval cells in which nuclear protein import is assured by the long persisting importin-beta molecules. How is the expression of the Ketel gene regulated? In silico analysis revealed several conserved transcription factor binding sequences in the Ketel gene promoter. Reporter genes in which different segments of the promoter ensured transient expression of the luciferase gene in S2 cells identified the sequences required for normal Ketel gene expression level. Gel retardation and band shift assays revealed that the DREF and the CFDD transcription factors play key roles in the regulation of Ketel gene expression. Transgenic LacZ reporter genes revealed the sequences that ensure tissue-specific gene expression. Apparently, the regulation of Ketel gene expression depends largely on a DRE motif and action of the DREF, CFDD, CF2-II and BEAF transcription factors.
在果蝇中由Ketel基因编码的输入蛋白β是核蛋白输入、纺锤体微管形成和核膜组装的关键组成部分。果蝇胚胎在生命开始时依赖母体提供的输入蛋白β。合子Ketel基因的表达在原肠胚形成期间在每个细胞中开始,并且虽然该表达在有丝分裂活跃的二倍体细胞中得以维持,但在不分裂的幼虫细胞中停止,在这些细胞中核蛋白输入由长期存在的输入蛋白β分子保证。Ketel基因的表达是如何调控的?计算机分析揭示了Ketel基因启动子中几个保守的转录因子结合序列。其中启动子不同片段确保荧光素酶基因在S2细胞中瞬时表达的报告基因确定了正常Ketel基因表达水平所需的序列。凝胶阻滞和条带迁移分析表明,DREF和CFDD转录因子在Ketel基因表达调控中起关键作用。转基因LacZ报告基因揭示了确保组织特异性基因表达的序列。显然,Ketel基因表达的调控很大程度上取决于一个DRE基序以及DREF、CFDD、CF2-II和BEAF转录因子的作用。