Bevilacqua A, Mangia F
Department of Psychology, La Sapienza University of Rome, Italy.
Dev Genet. 1993;14(2):92-102. doi: 10.1002/dvg.1020140203.
After fertilization in the mouse, the zygotic genome is activated in two-cell embryos by the spontaneous expression, among other genes, of the major inducible heat shock gene, hsp68, in the absence of heat-inducibility of heat shock genes. To obtain information on this phenomenon, we have probed one- and two-cell embryo's ability to express microinjected reporter DNA constructs, containing the Escherichia coli lacZ gene driven by promoters from early SV40 genes, the human beta-actin gene, and the normal or HSE-deleted mouse hsp68 gene. Activity of these promoters was also tested in mouse granulosa cells and dictyate oocytes, as a function of circular/linear construct configuration and occurrence of heat shock. The hsp68 promoter was heat-inducible in both granulosa cells and oocytes. Its heat activation required the presence of HSEs and, in the oocytes, of construct linear configuration. In the embryos however, this promoter was expressed independently of the presence of HSEs and of construct configuration, and its activity was not affected by heat shock. When constructs with early SV40 and beta-actin promoters were injected into one-cell embryos, they appeared to be inactivated with the first embryonic cleavage, in agreement with previous observations [Wiekowski et al., 1992]. By contrast, both normal and HSE-deleted hsp68 promoters maintained their activity through the first cleavage, providing the first evidence of a gene escaping such transcriptional repression. Present results confirm previous findings on hsp68 expression during early mouse development, and suggest that this activation is mediated by a factor(s) other than HSF.
在小鼠受精后,合子基因组在二细胞胚胎中被激活,这是通过主要的可诱导热休克基因hsp68等基因的自发表达实现的,此时热休克基因不存在热诱导性。为了获取有关这一现象的信息,我们探究了单细胞和二细胞胚胎表达显微注射的报告基因DNA构建体的能力,这些构建体包含由早期SV40基因启动子、人β-肌动蛋白基因以及正常或缺失热休克元件(HSE)的小鼠hsp68基因驱动的大肠杆菌lacZ基因。还在小鼠颗粒细胞和双线期卵母细胞中测试了这些启动子的活性,作为环形/线性构建体构型和热休克发生情况的函数。hsp68启动子在颗粒细胞和卵母细胞中均可被热诱导。其热激活需要热休克元件的存在,并且在卵母细胞中需要构建体的线性构型。然而,在胚胎中,该启动子的表达独立于热休克元件的存在和构建体构型,并且其活性不受热休克的影响。当将带有早期SV40和β-肌动蛋白启动子的构建体注射到单细胞胚胎中时,它们似乎在第一次胚胎分裂时失活,这与先前的观察结果一致[Wiekowski等人,1992]。相比之下,正常和缺失热休克元件的hsp68启动子在第一次分裂过程中均保持其活性,这为一个基因逃避这种转录抑制提供了首个证据。目前的结果证实了先前关于小鼠早期发育过程中hsp68表达的发现,并表明这种激活是由热休克因子(HSF)以外的一种或多种因子介导的。