Sequence changes preceding a Shine-Dalgarno region influence trpE mRNA translation and decay.
Sequence changes preceding a Shine-Dalgarno region influence trpE mRNA translation and decay.
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Shine-Dalgarno 区域之前的序列变化影响 trpE mRNA 翻译和衰减。
DOI：
10.1016/0022-2836(88)90598-0
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发表时间：
1988
影响因子：
5.6
通讯作者：
Yanofsky,C
中科院分区：
文献类型：
--
作者：
Cho,KO;Yanofsky,C
在使用atrpE启动子强度测量系统的研究中，我们观察到含有大肠杆菌trp启动子及其邻近转录区域的构建体产生比预期水平更低的trpE蛋白水平。为了分析这一观察结果，我们在 trpEShine-Dalgarno 区域之前的核苷酸序列中引入了突变变化，并检查了它们对 trpEmRNA 合成、翻译和衰变的影响。我们发现前 Shine-Dalgarno 区域中的某些缺失、插入和取代突变导致 intrpEenzyme 活性增加两到五倍。这些增加伴随着trpEmRNA稳态水平的增加。 trpEmRNA 降解的脉冲追踪分析表明，观察到的稳态 trpEmRNA 水平与 trpEmRNA 稳定性的变化相关。这些发现可以用替代模型来解释，其中提高 trpE 表达的突变变化的主要作用是增加 trpEmRNA 翻译，而不是增加 trpEmRNA 稳定性。
In studies with atrpEpromoter-strength measuring System we observed that constructs containing theEscherichia coli trppromoter and its adjacent transcribed region yielded lower levels oftrpEprotein than were expected. To analyze this observation we introduced mutational changes in the nucleotide sequence preceding thetrpEShine-Dalgarno region and examined their effects ontrpEmRNA synthesis, translation and decay. We found that certain deletion, insertion and Substitution mutations in the pre-Shine-Dalgarno region caused a two- to fivefold increase intrpEenzyme activity. These increases were accompanied by increases in steady-state levels oftrpEmRNA. Pulse-chase analyses oftrpEmRNA degradation revealed that the observed steady-statetrpEmRNA levels correlated with changes intrpEmRNA stability. These findings are interpreted in terms of alternative models in which the primary effect of mutational changes that elevatetrpEexpression is to increasetrpEmRNA translation,versusincreasingtrpEmRNA stability.