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Enhancement of translation by the epsilon element is independent of thesequence of the 460 region of 16S rRNA

机译:ε元件的翻译增强与16S rRNA 460区的序列无关

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摘要

The epsilon enhancer element is a pyrimidine-rich sequence that increases expression of T7 gene 10 and a number of Escherichia coli mRNAs during initiation of translation and inhibits expression of the recF mRNA during elongation. Based on its complementarity to the 460 region of 16S rRNA, it has been proposed that epsilon exerts its enhancer activity by base pairing to this complementary rRNA sequence. We have tested this model of enhancer action by constructing mutations in the 460 region of 16S rRNA and examining expression of epsilon-containing CAT reporter genes and recF-lacZ fusions in strains expressing the mutant rRNAs. Replacement of the 460 E. coli stem-loop with that of Salmonella enterica serovar Typhimurium or a stem-loop containing a reversal of all 8 bp in the helical region produced fully functional rRNAs with no apparent effect on cell growth or expression of any epsilon-containing mRNA, Our experiments confirm the reported effects of the epsilon elements on gene expression but show that these effects are independent of the sequence of the 460 region of 16S rRNA, indicating that epsilon-rRNA base pairing does not occur.
机译:ε增强子元件是富含嘧啶的序列,其在翻译起始期间增加T7基因10的表达和许多大肠杆菌mRNA的表达,并在延伸过程中抑制recF mRNA的表达。基于其与16S rRNA 460区的互补性,已提出ε通过与该互补rRNA序列碱基配对发挥其增强子活性。我们已经通过在16S rRNA的460个区域中构建突变并检查了表达突变rRNA的菌株中含ε的CAT报告基因和recF-lacZ融合蛋白的表达来测试了这种增强子作用模型。用小肠沙门氏菌血清鼠伤寒沙门氏菌替代460个大肠杆菌茎环或在螺旋区中包含所有8 bp逆转的茎环产生功能齐全的rRNA,对细胞生长或任何ε-表达均无明显影响我们的实验证实了含有εmRNA的ε元件对基因表达的影响,但表明这些作用与16S rRNA 460区的序列无关,表明ε-rRNA碱基配对并未发生。

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