Poly(A) polymerase is required for RyhB sRNA stability and function in Escherichia coli.
Poly(A) polymerase is required for RyhB sRNA stability and function in Escherichia coli.
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DOI：
10.1261/rna.067181.118
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发表时间：
2018-11
期刊：
影响因子：
--
通讯作者：
De Lay NR
中科院分区：
文献类型：
--
作者：
Sinha D;Matz LM;Cameron TA;De Lay NR
小分子调控RNA（Small regulatory RNA，sRNA）是一类重要的细菌转录后调节因子，控制着包括应激反应在内的多种生理过程。在革兰氏阴性细菌（包括大肠杆菌）中，RNA分子伴侣Hfq结合许多sRNA并促进与靶转录物的配对，导致mRNA转录、翻译或稳定性的变化。在这里，我们报告说，聚（A）聚合酶（PAP I），它促进RNA降解的外切核糖核酸酶通过添加聚（A）尾巴，在基因表达的调控Hfq依赖的sRNA中具有至关重要的作用。具体来说，我们发现，缺失pcnB，编码PAP I，矛盾地导致增加营业额的某些Hfq依赖的sRNA，包括RyhB。pcnB缺失菌株中RyhB的不稳定性受到以下因素的抑制：hfq或ryhB中破坏RyhB与靶RNA配对的突变;参与与RyhB配对的glyW-cysT-leuZ转录物（3′ETSLeuZ）3′外部转录间隔区的突变;或编码核糖核酸内切酶RNase E的rne的内部缺失。最后，RyhB在pcnB缺失菌株中的稳定性降低导致其一些靶mRNA的调节受损，特别是sodB和sdhCDAB。总的来说，我们的数据支持一个模型，其中PAP I在确保3′ETSLeuZ的有效衰变中起着关键作用。在没有PAP I的情况下，3′ETSLeuZ转录物积累，结合Hfq，并与RyhB配对，导致其通过RNase E介导的衰变而耗尽。这最终导致PAP I缺陷型菌株中RyhB功能的缺陷。
Small regulatory RNAs (sRNAs) are an important class of bacterial post-transcriptional regulators that control numerous physiological processes, including stress responses. In Gram-negative bacteria including Escherichia coli, the RNA chaperone Hfq binds many sRNAs and facilitates pairing to target transcripts, resulting in changes in mRNA transcription, translation, or stability. Here, we report that poly(A) polymerase (PAP I), which promotes RNA degradation by exoribonucleases through the addition of poly(A) tails, has a crucial role in the regulation of gene expression by Hfq-dependent sRNAs. Specifically, we show that deletion of pcnB, encoding PAP I, paradoxically resulted in an increased turnover of certain Hfq-dependent sRNAs, including RyhB. RyhB instability in the pcnB deletion strain was suppressed by mutations in hfq or ryhB that disrupt pairing of RyhB with target RNAs, by mutations in the 3′ external transcribed spacer of the glyW-cysT-leuZ transcript (3′ETSLeuZ) involved in pairing with RyhB, or an internal deletion in rne, which encodes the endoribonuclease RNase E. Finally, the reduced stability of RyhB in the pcnB deletion strain resulted in impaired regulation of some of its target mRNAs, specifically sodB and sdhCDAB. Altogether our data support a model where PAP I plays a critical role in ensuring the efficient decay of the 3′ETSLeuZ. In the absence of PAP I, the 3′ETSLeuZ transcripts accumulate, bind Hfq, and pair with RyhB, resulting in its depletion via RNase E-mediated decay. This ultimately leads to a defect in RyhB function in a PAP I deficient strain.
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