Requirement of upstream Hfq-binding (ARN)x elements in glmS and the Hfq C-terminal region for GlmS upregulation by sRNAs GlmZ and GlmY.
Requirement of upstream Hfq-binding (ARN)x elements in glmS and the Hfq C-terminal region for GlmS upregulation by sRNAs GlmZ and GlmY.
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DOI：
10.1093/nar/gks392
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发表时间：
2012-09
影响因子：
14.9
通讯作者：
Feig AL
中科院分区：
文献类型：
--
作者：
Salim NN;Faner MA;Philip JA;Feig AL
Hfq是一种重要的RNA结合蛋白，帮助细菌适应压力。其主要功能是促进反式作用小非编码RNA（sRNA）与其靶mRNA之间的配对。在rpoS和fhlA的上游区域中识别必需的Hfq结合基序使我们提出这样一个问题，即这些元件是否也在其他Hfq依赖性mRNA中常见。在这里，我们证实了glmS RNA中存在类似的（ARN）x基序，glmS RNA是由两种sRNA（GlmZ和GlmY）以Hfq依赖方式控制的基因。GlmZ代表典型的sRNA：mRNA配对系统，而GlmY是非典型的，与RNA加工蛋白YhbJ相互作用。我们发现，glmS相互作用的两个Hfq结合表面的sRNA的情况下。即使存在两个（ARN）x基序，使用glmS：gfp融合系统，我们确定只有一个特定的（ARN）x元素是必不可少的调节。此外，我们表明，在大肠杆菌Hfq的C-末端延伸残基66-72是必不可少的激活GlmS表达的GlmY，但不与GlmZ。这一结果表明，Hfq的C-末端延伸可能是涉及辅助组分如额外的RNA或蛋白质的某些形式的非典型sRNA调节所需的。
Hfq is an important RNA-binding protein that helps bacteria adapt to stress. Its primary function is to promote pairing between trans-acting small non-coding RNAs (sRNAs) and their target mRNAs. Identification of essential Hfq-binding motifs in up-stream regions of rpoS and fhlA led us to ask the question whether these elements are a common occurrence among other Hfq-dependent mRNAs as well. Here, we confirm the presence of a similar (ARN)x motif in glmS RNA, a gene controlled by two sRNAs (GlmZ and GlmY) in an Hfq-dependent manner. GlmZ represents a canonical sRNA:mRNA pairing system, whereas GlmY is non-canonical, interfacing with the RNA processing protein YhbJ. We show that glmS interacts with both Hfq-binding surfaces in the absence of sRNAs. Even though two (ARN)x motifs are present, using a glmS:gfp fusion system, we determined that only one specific (ARN)x element is essential for regulation. Furthermore, we show that residues 66–72 in the C-terminal extension of Escherichia coli Hfq are essential for activation of GlmS expression by GlmY, but not with GlmZ. This result shows that the C-terminal extension of Hfq may be required for some forms of non-canonical sRNA regulation involving ancillary components such as additional RNAs or proteins.
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