Molina A, Di Martino E, Martial J A, Muller M
Laboratoire de Biologie Moléculaire et Génie Génétique, Université de Liège, Institut de Chimie B6, B-4000 Sart-Tilman, Belgium.
Biochem J. 2001 Jun 1;356(Pt 2):353-9. doi: 10.1042/0264-6021:3560353.
We reported previously that a tilapia (Oreochromis mossambicus) heat shock protein 70 (HSP70) promoter is able to confer heat shock response on a reporter gene after transient expression both in cell culture and in microinjected zebrafish embryos. Here we present the first functional analysis of a fish HSP70 promoter, the tiHSP70 promoter. Using transient expression experiments in carp EPC (epithelioma papulosum cyprini) cells and in microinjected zebrafish embryos, we show that a distal heat shock response element (HSE1) at approx. -800 is predominantly responsible for the heat shock response of the tiHSP70 promoter. This element specifically binds an inducible transcription factor, most probably heat shock factor, and a constitutive factor. The constitutive complex is not observed with the non-functional, proximal HSE3 sequence, suggesting that both factors are required for the heat shock response mediated by HSE1.
我们之前报道过,罗非鱼(莫桑比克罗非鱼)热休克蛋白70(HSP70)启动子在细胞培养和显微注射的斑马鱼胚胎中瞬时表达后,能够赋予报告基因热休克反应。在此,我们展示了首个鱼类HSP70启动子——tiHSP70启动子的功能分析。通过在鲤鱼EPC(鲤上皮瘤)细胞和显微注射的斑马鱼胚胎中进行瞬时表达实验,我们发现大约位于-800处的一个远端热休克反应元件(HSE1)主要负责tiHSP70启动子的热休克反应。该元件特异性结合一种诱导型转录因子,很可能是热休克因子,以及一种组成型因子。在无功能的近端HSE3序列中未观察到组成型复合物,这表明这两种因子都是HSE1介导的热休克反应所必需的。