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Species-specificity of rRNA gene transcription in plants manifested as a switch in RNA polymerase specificity

机译:植物中rRNA基因转录的物种特异性表现为RNA聚合酶特异性的转换

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摘要

Rapid evolution of ribosomal RNA (rRNA) gene promoters often prevents their recognition in a foreign species. Unlike animal systems, we show that foreign plant rRNA gene promoters are recognized in an alien species, but tend to program transcription by a different polymerase. In plants, RNA polymerase 1 transcripts initiate at a TATATA element (+1 is underlined) important for promoter strength and start-site selection. However, transcripts initiate from +32 following transfection of a tomato promoterinto Arabidopsis. The rRNA gene promoter of a more closely related species, Brassica oleracea, programs both +1 and +29 transcription. A point mutation at +2 improving the identity between the Brassica and Arabidopsis promoters increases +1 transcription, indicating a role for the initiator element in species-specificity. Brassica +29 transcripts can be translated to express a luciferase reporter gene, implicating RNA polymerase II. TATA mutations that disrupt TATA-binding protein (TBP) interactions inhibit +29 transcription and luciferase expression. Co-expressed TBP proteins bearing compensatory mutations restore +29 transcription and luciferase activity, suggesting a direct TBP-TATA interaction. Importantly, +1 transcription is unaffected by the TATA mutations, suggesting that in the context of pol I recognition, the TATA-containing initiator element serves a function other than TBP binding.
机译:核糖体RNA(rRNA)基因启动子的快速进化通常会阻止它们在外来物种中的识别。与动物系统不同,我们显示外来植物rRNA基因启动子在外来物种中被识别,但是倾向于通过不同的聚合酶来编程转录。在植物中,RNA聚合酶1转录物起始于TATATA元件(带下划线的+1)对启动子强度和起始位点选择很重要。然而,在番茄启动子转染到拟南芥中后,转录物从+32开始。密切相关的物种芸苔属的rRNA基因启动子可对+1和+29转录进行编程。 +2处的一个点突变改善了芸苔和拟南芥启动子之间的同一性,从而增加了+1转录,表明启动子元件在物种特异性中的作用。可以将芸苔属+29转录本翻译成荧光素酶报道基因,暗示RNA聚合酶II。破坏TATA结合蛋白(TBP)相互作用的TATA突变抑制+29转录和荧光素酶表达。带有补偿性突变的共表达TBP蛋白可恢复+29转录和荧光素酶活性,表明TBP-TATA直接相互作用。重要的是,+ 1转录不受TATA突变的影响,这表明在pol I识别的背景下,含TATA的引发剂元件具有TBP结合以外的功能。

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