USDA-ARS Appalachian Fruit Research Station, 2217 Wiltshire Road, Kearneysville, WV 25430, USA.
Plant Mol Biol. 2010 Oct;74(3):293-305. doi: 10.1007/s11103-010-9673-9. Epub 2010 Aug 12.
The expression of eukaryotic genes from their cognate promoters is often regulated by the action of transcriptional enhancer elements that function in an orientation-independent manner either locally or at a distance within a genome. This interactive nature often provokes unexpected interference within transgenes in plants, as reflected by misexpression of the introduced gene and undesired phenotypes in transgenic lines. To gain a better understanding of the mechanism underlying enhancer/promoter interactions in a plant system, we analyzed the activation of a β-glucuronidase (GUS) reporter gene by enhancers contained within the AGAMOUS second intron (AGI) and the Cauliflower Mosaic Virus (CaMV) 35S promoter, respectively, in the presence and absence of a target promoter. Our results indicate that both the AGI and 35S enhancers, which differ significantly in their species of origin and in the pattern of expression that they induce, have the capacity to activate the expression of a nearby gene through the promoter-independent initiation of autonomous transcriptional events. Furthermore, we provide evidence that the 35S enhancer utilizes a mechanism resembling animal- and yeast-derived scanning or facilitated tracking models of long-distance enhancer action in its activation of a remote target promoter.
真核基因从其同源启动子表达通常受到转录增强子元件的调控,这些元件以非定向的方式在基因组内的局部或远距离发挥作用。这种相互作用的性质经常在植物中转基因中引起意想不到的干扰,表现为引入基因的错误表达和转基因系中出现不需要的表型。为了更好地理解植物系统中增强子/启动子相互作用的机制,我们分析了 AGAMOUS 第二内含子(AGI)内的增强子和 Cauliflower Mosaic Virus(CaMV)35S 启动子分别在存在和不存在靶启动子时对 β-葡萄糖醛酸酶(GUS)报告基因的激活作用。我们的结果表明,AGI 和 35S 增强子在其起源物种和诱导表达模式上有很大的不同,它们都有能力通过自主转录事件的启动来激活附近基因的表达,而无需启动子。此外,我们提供的证据表明,35S 增强子利用一种机制,类似于动物和酵母衍生的扫描或促进跟踪模型,在其激活远程靶启动子时发挥长距离增强子作用。