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Inactivation of RNase P in Escherichia coli significantly changes post-transcriptional RNA metabolism
Molecular Microbiology ( IF 3.6 ) Pub Date : 2021-09-06 , DOI: 10.1111/mmi.14808
Bijoy K Mohanty 1 , Sidney R Kushner 1, 2
Affiliation  

Ribonuclease P (RNase P), which is required for the 5'-end maturation of tRNAs in every organism, has been shown to play a limited role in other aspects of RNA metabolism in Escherichia coli. Using RNA-sequencing (RNA-seq), we demonstrate that RNase P inactivation affects the abundances of ~46% of the expressed transcripts in E. coli and provide evidence that its essential function is its ability to generate pre-tRNAs from polycistronic tRNA transcripts. The RNA-seq results agreed with the published data and northern blot analyses of 75/83 transcripts (mRNAs, sRNAs, and tRNAs). Changes in transcript abundances in the RNase P mutant also correlated with changes in their half-lives. Inactivating the stringent response did not alter the rnpA49 phenotype. Most notably, increases in the transcript abundances were observed for all genes in the cysteine regulons, multiple toxin-antitoxin modules, and sigma S-controlled genes. Surprisingly, poly(A) polymerase (PAP I) modulated the abundances of ~10% of the transcripts affected by RNase P. A comparison of the transcriptomes of RNase P, RNase E, and RNase III mutants suggests that they affect distinct substrates. Together, our work strongly indicates that RNase P is a major player in all aspects of post-transcriptional RNA metabolism in E. coli.

中文翻译:

大肠杆菌中 RNase P 的失活显着改变转录后 RNA 代谢

核糖核酸酶 P (RNase P) 是每个生物体中 tRNA 5' 端成熟所必需的,已被证明在大肠杆菌中RNA 代谢的其他方面发挥的作用有限。使用 RNA 测序 (RNA-seq),我们证明 RNase P 失活影响大肠杆菌中约 46% 的表达转录本的丰度,并提供证据表明其基本功能是从多顺反子 tRNA 转录本生成前 tRNA 的能力. RNA-seq 结果与已发表的数据和 75/83 转录本(mRNA、sRNA 和 tRNA)的 northern 印迹分析一致。RNase P 突变体中转录物丰度的变化也与其半衰期的变化相关。使严格反应失活并没有改变rnpA49表型。最值得注意的是,观察到半胱氨酸调节子、多重毒素-抗毒素模块和 sigma S 控制基因中所有基因的转录丰度增加。令人惊讶的是,poly(A) 聚合酶 (PAP I) 调节了约 10% 受 RNase P 影响的转录本的丰度。RNase P、RNase E 和 RNase III 突变体的转录组比较表明它们影响不同的底物。总之,我们的工作有力地表明 RNase P 在大肠杆菌转录后 RNA 代谢的各个方面都起着重要作用。
更新日期:2021-09-06
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