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Hfq affects mRNA levels independently of degradation

机译:Hfq独立于降解而影响mRNA水平

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摘要

Background The bacterial Lsm protein, Hfq, is an RNA chaperone involved in many reactions related to RNA metabolism, such as replication and stability, control of small RNA activity and polyadenylation. Despite this wide spectrum of known functions, the global role of Hfq is almost certainly undervalued; its capacity to bind DNA and to interact with many other proteins are only now beginning to be taken into account.Results The role of Hfq in the maturation and degradation of the rpsO mRNA of E. coli was investigated in vivo. The data revealed a decrease in rpsO mRNA abundance concomitant to an increase in its stability when Hfq is absent. This indicates that the change in mRNA levels in hfq mutants does not result from its modification of RNA stability. Moreover, a series of independent experiments have revealed that the decrease in mRNA level is not a consequence of a reduction of translation efficiency and that Hfq is not directly implicated in translational control of rpsO expression. Reduced steady-state mRNA levels in the absence of Hfq were also shown for rpsT, rpsB and rpsB-tsf, but not for lpp, pnp or tRNA transcripts. The abundance of chimeric transcripts rpsO-lacZ and rpsB-lacZ, whose expression was driven by rpsO and rpsB promoters, respectively, was also lower in the hfq null-mutants, while the β-galactosidase yield remained about the same as in the parent wild-type strain.Conclusions The data obtained suggest that alteration of rpsO, rpsT and rpsB-tsf transcript levels observed under conditions of Hfq deficiency is not caused by the post-transcriptional events, such as mRNA destabilization or changes in translation control, and may rather result from changes in transcriptional activity. So far, how Hfq affects transcription remains unclear. We propose that one of the likely mechanisms of Hfq-mediated modulation of transcription might operate early in the elongation step, when interaction of Hfq with a nascent transcript would help to overcome transcription pauses and to prevent preliminary transcript release.
机译:背景技术细菌Lsm蛋白Hfq是一种RNA伴侣,参与许多与RNA代谢相关的反应,例如复制和稳定性,小RNA活性的控制和聚腺苷酸化。尽管已知功能种类繁多,但Hfq的全球作用几乎可以肯定被低估了;直到现在才开始考虑其结合DNA和与许多其他蛋白质相互作用的能力。结果在体内研究了Hfq在大肠杆菌rpsO mRNA成熟和降解中的作用。数据显示,当缺乏Hfq时,rpsO mRNA丰度降低,同时稳定性增加。这表明hfq突变体中mRNA水平的变化不是由其RNA稳定性的改变引起的。此外,一系列独立的实验表明,mRNA水平的降低不是翻译效率降低的结果,并且Hfq与rpsO表达的翻译控制没有直接关系。对于rpsT,rpsB和rpsB-tsf,在没有Hfq的情况下,稳态mRNA水平也降低,但对于lpp,pnp或tRNA转录本,则没有降低。分别由rpsO和rpsB启动子驱动表达的嵌合转录本rpsO-lacZ和rpsB-lacZ的丰度在hfq null突变体中也较低,而β-半乳糖苷酶的产量与亲本野生株保持大致相同。结论所获得的数据表明,在Hfq缺乏的条件下观察到的rpsO,rpsT和rpsB-tsf转录水平的改变不是由转录后事件引起的,例如mRNA不稳定或翻译控制的改变,而可能是由转录活性的变化引起。到目前为止,Hfq如何影响转录仍不清楚。我们提出,Hfq介导的转录调控的可能机制之一可能在延伸步骤的早期起作用,此时Hfq与新生转录本的相互作用将有助于克服转录停顿并防止初步转录本的释放。

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