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首页> 美国卫生研究院文献>The EMBO Journal >Mechanism of post-segregational killing: Sok antisense RNA interacts with Hok mRNA via its 5-end single-stranded leader and competes with the 3-end of Hok mRNA for binding to the mok translational initiation region.
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Mechanism of post-segregational killing: Sok antisense RNA interacts with Hok mRNA via its 5-end single-stranded leader and competes with the 3-end of Hok mRNA for binding to the mok translational initiation region.

机译：分离后杀死的机制：Sok反义RNA通过其5端单链前导物与Hok mRNA相互作用并与Hok mRNA的3末端竞争与mok翻译起始区的结合。

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摘要

The hok/sok system of plasmid R1, which mediates plasmid stabilization by killing of plasmid-free segregants, codes for two RNA species, Hok mRNA and Sok antisense RNA. The lethal expression of hok is inhibited post-transcriptionally by the 67 nt Sok-RNA. In this paper, we analyse the secondary structure of Sok-RNA and the binding of Sok-RNA to Hok mRNA in vitro. The reaction between the two RNAs leads to the formation of a complete duplex in which Sok-RNA is hybridized over its entire length to Hok mRNA. The second-order rate constant of duplex formation was determined to be approximately 1 x 10(5) M-1s-1. Mutations in the 5'-end single-stranded leader of Sok-RNA severely reduced the binding rate to wt Hok mRNA, whereas loop mutations in Sok-RNA had no such effect. The reduced binding rates were paralleled by abolished in vivo regulatory properties. These results suggest that, unlike in other well-characterized antisense/target RNA systems, the initial recognition reaction between Sok-RNA and Hok mRNA takes place between the single-stranded 5'-end of Sok-RNA and the complementary region in Hok mRNA, without the involvement of an antisense loop in the initial binding step. Furthermore, the finding that Sok-RNA competes with the 3'-end of full-length Hok mRNA for binding to the mok translational initiation region adds to the complexity of killer gene regulation.
机译：质粒R1的hok / sok系统通过杀死无质粒的分离子来介导质粒稳定，编码两种RNA，即Hok mRNA和Sok反义RNA。转录后的67 nt Sok-RNA抑制了hok的致死表达。在本文中，我们分析了Sok-RNA的二级结构以及Sok-RNA与Hok mRNA的结合。两个RNA之间的反应导致形成完整的双链体，其中Sok-RNA在其全长上与Hok mRNA杂交。确定双链体形成的二级速率常数约为1 x 10（5）M-1s-1。 Sok-RNA的5'端单链前导序列中的突变严重降低了与wt Hok mRNA的结合率，而Sok-RNA中的环突变则没有这种作用。降低的结合速率与消除的体内调节特性平行。这些结果表明，与其他特征明​​确的反义/靶标RNA系统不同，Sok-RNA和Hok mRNA之间的初始识别反应发生在Sok-RNA的单链5'末端与Hok mRNA的互补区域之间，而在初始绑定步骤中不涉及反义环。此外，Sok-RNA与全长Hok mRNA的3'末端竞争与mok翻译起始区域结合的发现增加了杀手基因调控的复杂性。

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