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首页> 外文期刊>Journal of bacteriology >CsrA Represses Translation of sdiA, Which Encodes the N-Acylhomoserine-l-Lactone Receptor of Escherichia coli, by Binding Exclusively within the Coding Region of sdiA mRNA
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CsrA Represses Translation of sdiA, Which Encodes the N-Acylhomoserine-l-Lactone Receptor of Escherichia coli, by Binding Exclusively within the Coding Region of sdiA mRNA

机译：CsrA抑制sdiA的翻译，该编码仅在sdiA mRNA的编码区域内结合，从而编码大肠杆菌的N-酰基高丝氨酸-1-内酯受体。

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The RNA binding protein CsrA is the central component of a conserved global regulatory system that activates or represses gene expression posttranscriptionally. In every known example of CsrA-mediated translational control, CsrA binds to the 5′ untranslated region of target transcripts, thereby repressing translation initiation and/or altering the stability of the RNA. Furthermore, with few exceptions, repression by CsrA involves binding directly to the Shine-Dalgarno sequence and blocking ribosome binding. sdiA encodes the quorum-sensing receptor for N-acyl-l-homoserine lactone in Escherichia coli. Because sdiA indirectly stimulates transcription of csrB, which encodes a small RNA (sRNA) antagonist of CsrA, we further explored the relationship between sdiA and the Csr system. Primer extension analysis revealed four putative transcription start sites within 85 nucleotides of the sdiA initiation codon. Potential σ70-dependent promoters were identified for each of these primer extension products. In addition, two CsrA binding sites were predicted in the initially translated region of sdiA. Expression of chromosomally integrated sdiA′-′lacZ translational fusions containing the entire promoter and CsrA binding site regions indicates that CsrA represses sdiA expression. The results from gel shift and footprint studies demonstrate that tight binding of CsrA requires both of these sites. Furthermore, the results from toeprint and in vitro translation experiments indicate that CsrA represses translation of sdiA by directly competing with 30S ribosomal subunit binding. Thus, this represents the first example of CsrA preventing translation by interacting solely within the coding region of an mRNA target.
机译：RNA结合蛋白CsrA是保守的全局调节系统的核心组件，该系统在转录后激活或抑制基因表达。在CsrA介导的翻译控制的每个已知实例中，CsrA结合至目标转录本的5'非翻译区，从而抑制翻译起始和/或改变RNA的稳定性。此外，除少数例外，CsrA抑制涉及直接与Shine-Dalgarno序列结合并阻断核糖体结合。 sdiA 编码大肠杆菌中 N -酰基-1-高丝氨酸内酯的群体感应受体。由于 sdiA 间接刺激了 csrB 的转录，而该编码是CsrA的小RNA（sRNA）拮抗剂，因此我们进一步探讨了 sdiA 与CsrA的关系。企业社会责任系统。引物延伸分析显示在 sdiA 起始密码子的85个核苷酸内的四个假定的转录起始位点。对于这些引物延伸产物中的每一个，都鉴定出潜在的依赖于σ 70 的启动子。另外，在 sdiA 的最初翻译区域中预测到两个CsrA结合位点。包含完整启动子和CsrA结合位点区域的染色体整合的 sdiA '-' lacZ 翻译融合体的表达表明CsrA抑制 sdiA 表达。凝胶迁移和印迹研究的结果表明，CsrA的紧密结合需要这两个位点。此外，脚印和体外翻译实验的结果表明，CsrA通过直接与30S核糖体亚基结合竞争而抑制 sdiA 的翻译。因此，这代表了CsrA通过仅在mRNA靶标的编码区内相互作用而阻止翻译的第一个例子。

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