Functional Escherichia coli 23S rRNAs containing processed and unprocessed intervening sequences from Salmonella typhimurium
Functional Escherichia coli 23S rRNAs containing processed and unprocessed intervening sequences from Salmonella typhimurium
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DOI：
10.1093/nar/24.24.4918
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发表时间：
1996-12-15
影响因子：
14.9
通讯作者：
Dahlberg, AE
中科院分区：
文献类型：
--
作者：
Gregory, ST;OConnor, M;Dahlberg, AE
我们将鼠伤寒沙门氏菌rrnD操纵子的23S rRNA中的干扰序列(IVS)引入到大肠杆菌23S rRNA的等位基因中，鼠伤寒沙门氏菌23S rRNA由于依赖RNaseIII去除了组成IVS的类似100个核苷酸的茎环结构而碎片化，在本研究中，我们发现将鼠伤寒沙门氏菌的IVS插入到E.Coli23S rRNA中会导致RNA的碎片化，但不影响核糖体的功能，表达碎片化的23S rRNA的细胞呈现野生型生长速度，在活跃翻译的核糖池中发现了碎片化的RNA，但不改变核糖体亚单位的沉淀谱，70S糖体或多聚体携带A2058G突变的杂交23S rRNA赋予红霉素高水平的耐药性，与携带该突变的完整23S rRNA的抗性无法区分，这些观察表明，这种IVS的存在及其去除是表型沉默的，正如在RNaseIII缺陷株中观察到的那样，IVS的处理不是产生功能性核糖体所必需的。
We have introduced the intervening sequence (IVS) from 23S rRNA of the rrnD operon of Salmonella typhimurium into the equivalent position of Escherichia coli 23S rRNA, Salmonella typhimurium 23S rRNA is fragmented due to the RNase III-dependent removal of the similar to 100 nt stem-loop structure that comprises the IVS, In this study, we have found that insertion of the S.typhimurium IVS into E.coli 23S rRNA causes fragmentation of the RNA but does not affect ribosome function, Cells expressing the fragmented 23S rRNA exhibited wild-type growth rates, Fragmented RNA was found in the actively translating polysome pool and did not alter the sedimentation profile of ribosomal subunits, 70S ribosomes or polysomes, Finally, hybrid 23S rRNA carrying the A2058G mutation conferred high level erythromycin resistance indistinguishable from that of intact 23S rRNA carrying this mutation, These observations indicate that the presence of this IVS and its removal are phenotypically silent, As observed in an RNase III-deficient strain, processing of the IVS was not required for the production of functional ribosomes.