Robbins P D, Rio D C, Botchan M R
Mol Cell Biol. 1986 Apr;6(4):1283-95. doi: 10.1128/mcb.6.4.1283-1295.1986.
We describe experiments which demonstrated that the simian virus 40 (SV40) enhancer affects certain transcriptional units differently. We also found that a specific enhancer-transcriptional unit interaction can be regulated by trans-acting factors. Using transient assays, we examined the effects of the SV40 enhancer on herpesvirus thymidine kinase (tk) RNA levels when transcription was initiated either by the herpesvirus tk promoter or by an SV40 early promoter-tk fusion. We were unable to detect any effect of the enhancer on transcription from the tk promoter in CV-1 or HeLa cells. However, we found that the addition of T-antigen in trans allowed the enhancer to stimulate expression from the tk promoter. This induction by T-antigen did not require T-antigen-binding sites in cis and appeared to be an indirect effect. In contrast, tk expression from the SV40 early promoter fusion was greatly stimulated by the enhancer in CV-1 cells. Furthermore, in 293 cells the SV40 enhancer had only a marginal effect on the SV40 promoter-tk fusion, whereas it strongly stimulated tk expression from the tk promoter. Our results raise the possibility that the enhancer function may not show cell specificity per se; rather, the interaction between the enhancer and a specific gene may be responsible for cell specificity. We discuss these observations in terms of the SV40 early gene-to-late gene switch that occurs during SV40 lytic growth.
我们描述了一些实验,这些实验表明猿猴病毒40(SV40)增强子对某些转录单位的影响有所不同。我们还发现,特定的增强子-转录单位相互作用可受反式作用因子调控。利用瞬时分析,我们检测了当转录由疱疹病毒胸苷激酶(tk)启动子或SV40早期启动子-tk融合体启动时,SV40增强子对疱疹病毒tk RNA水平的影响。我们在CV-1或HeLa细胞中未能检测到增强子对tk启动子转录的任何影响。然而,我们发现反式添加T抗原可使增强子刺激tk启动子的表达。T抗原的这种诱导作用不需要顺式的T抗原结合位点,似乎是一种间接效应。相比之下,在CV-1细胞中,SV40早期启动子融合体的tk表达受到增强子的极大刺激。此外,在293细胞中,SV40增强子对SV40启动子-tk融合体只有轻微影响,而它强烈刺激tk启动子的tk表达。我们的结果提出了一种可能性,即增强子功能本身可能不表现出细胞特异性;相反,增强子与特定基因之间的相互作用可能是细胞特异性的原因。我们根据SV40裂解生长过程中发生的SV40早期基因到晚期基因的转换来讨论这些观察结果。