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Hfq Is a Regulator of F-Plasmid TraJ and TraM Synthesis in Escherichia coli

机译:Hfq是大肠杆菌中F-质粒TraJ和TraM合成的调节剂

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摘要

The F plasmid of Escherichia coli allows horizontal DNA transfer between an F+ donor cell and an F recipient. Expression of the transfer genes is tightly controlled by a number of factors, including the following plasmid-encoded regulatory proteins: TraJ, the primary activator of the 33-kb tra operon, and the autoregulators TraM and TraY. Here, we demonstrate that the host RNA binding protein, Hfq, represses TraJ and TraM synthesis by destabilizing their respective mRNAs. Mating assays and immunoblot analyses for TraM and TraJ showed that transfer efficiency and protein levels increased in host cells containing a disruption in hfq compared to wild-type cells in stationary phase. The stability of transcripts containing a putative Hfq binding site located in the intergenic untranslated region between traM and traJ was increased in hfq mutant donor cells, suggesting that Hfq destabilizes these transcripts. Electrophoretic mobility shift assays demonstrated that Hfq specifically binds this region but not the antisense RNA, FinP, encoded on the opposite strand. Together, these findings indicate that Hfq regulates traM and traJ transcript stability by a mechanism separate from FinOP-mediated repression.
机译:大肠杆菌的F质粒允许F + 供体细胞和F -受体之间的水平DNA转移。转移基因的表达受到许多因素的严格控制,包括以下质粒编码的调控蛋白:TraJ,33 kb tra操纵子的主要激活剂以及自动调控剂TraM和TraY。在这里,我们证明了宿主RNA结合蛋白Hfq通过使各自的mRNA不稳定来抑制TraJ和TraM的合成。 TraM和TraJ的交配检测和免疫印迹分析表明,与固定相中的野生型细胞相比,包含hfq受到破坏的宿主细胞的转移效率和蛋白质水平提高了。在hfq突变体供体细胞中,包含位于traM和traJ之间的基因间非翻译区的推定Hfq结合位点的转录本的稳定性增加,表明Hfq使这些转录本不稳定。电泳迁移率变动分析表明,Hfq特异性结合该区域,但不结合反链上编码的反义RNA FinP。总之,这些发现表明,Hfq通过与FinOP介导的阻抑不同的机制来调节traM和traJ转录物的稳定性。

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