Adjacent single-stranded regions mediate processing of tRNA precursors by RNase E direct entry.
Adjacent single-stranded regions mediate processing of tRNA precursors by RNase E direct entry.
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相邻的单链区域通过RNase E直接进入介导TRNA前体的处理。
DOI：
10.1093/nar/gkt1403
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
McDowall KJ
中科院分区：
文献类型：
--
作者：
Kime L;Clarke JE;Romero A D;Grasby JA;McDowall KJ
RNase E家族因在许多细菌物种中对所有类型的RNA的加工和降解起中心作用而闻名，并且提供了可以识别5′-单磷酸化末端从而提高切割效率的核酸内切酶的第一个例子。然而，越来越多的证据表明，大肠杆菌RNase E可以通过直接进入，即在不识别5′-单磷酸化末端的情况下，有效地切割某些转录物。在这里，我们提供了生物化学证据，直接进入是中央处理的转移RNA（tRNA）在E。大肠杆菌中，RNase E的核心功能之一，并表明它是由特定的未配对区域介导的，这些区域与RNase E切割的片段相邻，但不连续。此外，我们发现直接进入tRNA前体5′侧的位点会触发一系列5′-单磷酸依赖性切割。与直接进入tRNA加工中的主要作用一致，我们提供了额外的证据表明，不需要5′-单磷酸来激活切割中的催化步骤。还提供了通过直接进入加工的tRNA前体的其他实例。因此，越来越多的证据表明，RNase E的直接进入在细菌RNA代谢中起着重要作用。
The RNase E family is renowned for being central to the processing and decay of all types of RNA in many species of bacteria, as well as providing the first examples of endonucleases that can recognize 5′-monophosphorylated ends thereby increasing the efficiency of cleavage. However, there is increasing evidence that some transcripts can be cleaved efficiently by Escherichia coli RNase E via direct entry, i.e. in the absence of the recognition of a 5′-monophosphorylated end. Here, we provide biochemical evidence that direct entry is central to the processing of transfer RNA (tRNA) in E. coli, one of the core functions of RNase E, and show that it is mediated by specific unpaired regions that are adjacent, but not contiguous to segments cleaved by RNase E. In addition, we find that direct entry at a site on the 5′ side of a tRNA precursor triggers a series of 5′-monophosphate-dependent cleavages. Consistent with a major role for direct entry in tRNA processing, we provide additional evidence that a 5′-monophosphate is not required to activate the catalysis step in cleavage. Other examples of tRNA precursors processed via direct entry are also provided. Thus, it appears increasingly that direct entry by RNase E has a major role in bacterial RNA metabolism.
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影响因子：
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