Mutational analysis of the L1 binding site of 23S rRNA in Escherichia coli.
Mutational analysis of the L1 binding site of 23S rRNA in Escherichia coli.
复制标题
大肠杆菌23S rRNA L1结合位点的突变分析。
DOI：
10.1093/nar/16.22.10529
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发表时间：
1988
影响因子：
14.9
通讯作者：
Nomura,M
中科院分区：
文献类型：
--
作者：
Said,B;Cole,JR;Nomura,M
大肠杆菌的L11核糖体蛋白操纵子含有L11和L1的基因，并受翻译阻遏物L1的反馈调节。23 S rRNA上的L1结合位点和L11操纵子mRNA上的L1阻遏物靶位点都具有相似的二级结构，并含有一些一级序列同一性。构建了23 S rRNA结合区的几个定点突变，并检查了它们对结合的影响。体外分析采用滤膜结合法。在体内分析中，使用条件表达系统过量产生含有L1结合区的23 S rRNA片段，这导致L11和L1合成的特异性去抑制。23 S rRNA L1结合位点的共享区域的变化对L1结合产生的影响类似于先前在L11操纵子mRNA靶位点的相应变化分析中发现的那些。结果支持的假设，r-蛋白L1相互作用与23 S rRNA和L11操纵子mRNA识别两种RNA上的相似功能。
The L11 ribosomal protein operon ofEscherichia colicontains the genes for L11 and L1 and is feedback regulated by the translational repressor L1. Both the L1 binding site on 23S rRNA and the L1 repressor target site on L11 operon mRNA share similar proposed secondary structures and contain some primary sequence identity. Several site-directed mutations in the binding region of 23S rRNA were constructed and their effects on binding were examined. Forin vitroanalysis, a filter binding method was used. Forin vivoanalysis, a conditional expression system was used to overproduce a 23S rRNA fragment containing the L1 binding region, which leads to specific derepression of the synthesis of L11 and L1. Changes in the shared region of the 23S rRNA L1 binding site produced effects on L1 binding similar to those found previously in analysis of corresponding changes in the L11 operon mRNA target site. The results support the hypothesis that r-protein L1 interacts with both 23S rRNA and L11 operon mRNA by recognizing similar features on both RNAs.