Direct entry by RNase E is a major pathway for the degradation and processing of RNA in Escherichia coli.
Direct entry by RNase E is a major pathway for the degradation and processing of RNA in Escherichia coli.
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DOI：
10.1093/nar/gku808
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发表时间：
2014-10
影响因子：
14.9
通讯作者：
McDowall KJ
中科院分区：
文献类型：
--
作者：
Clarke JE;Kime L;Romero A D;McDowall KJ
大肠杆菌核糖核酸内切酶E对基因表达有重要影响。它对核糖体和转移RNA的成熟以及信使RNA的快速降解至关重要。后者确保翻译在转录水平上紧跟编程。最近，RNase E的特征之一，即其通过5 ' -单磷酸化端结合的能力，被证明对一些多顺反电子tRNA前体的初始切割是不必要的。在这里，我们通过对体内核糖核酸酶缺陷菌株和体外RNase E的5 ' -传感器突变体的RNA-seq分析表明，与目前的模型相反，与5 ' -单磷酸盐无关的“直接进入”切割是降解和加工RNA的主要途径。此外，我们提供了进一步的证据，表明RNase E可以同时结合多个未配对区域，从而促进直接进入。这些简单的要求可以最大限度地提高降解和处理的速度，允许多个地点直接调查，而不受5 '端系绳的限制。在先前描述的RNase E的许多位点检测到切割，包括调节核糖体活性和特异性的位点。研究还揭示了RNase G（一种RNase E旁系物）在修剪5 ' -单磷酸化末端中的潜在广泛作用。
Escherichia coli endoribonuclease E has a major influence on gene expression. It is essential for the maturation of ribosomal and transfer RNA as well as the rapid degradation of messenger RNA. The latter ensures that translation closely follows programming at the level of transcription. Recently, one of the hallmarks of RNase E, i.e. its ability to bind via a 5′-monophosphorylated end, was shown to be unnecessary for the initial cleavage of some polycistronic tRNA precursors. Here we show using RNA-seq analyses of ribonuclease-deficient strains in vivo and a 5′-sensor mutant of RNase E in vitro that, contrary to current models, 5′-monophosphate-independent, ‘direct entry’ cleavage is a major pathway for degrading and processing RNA. Moreover, we present further evidence that direct entry is facilitated by RNase E binding simultaneously to multiple unpaired regions. These simple requirements may maximize the rate of degradation and processing by permitting multiple sites to be surveyed directly without being constrained by 5′-end tethering. Cleavage was detected at a multitude of sites previously undescribed for RNase E, including ones that regulate the activity and specificity of ribosomes. A potentially broad role for RNase G, an RNase E paralogue, in the trimming of 5′-monophosphorylated ends was also revealed.
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