Cukras Anthony R, Green Rachel
Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
J Mol Biol. 2005 May 27;349(1):47-59. doi: 10.1016/j.jmb.2005.03.075. Epub 2005 Apr 12.
The ribosomal protein S13 is found in the head region of the small subunit, where it interacts with the central protuberance of the large ribosomal subunit and with the P site-bound tRNA through its extended C terminus. The bridging interactions between the large and small subunits are dynamic, and are thought to be critical in orchestrating the molecular motions of the translation cycle. S13 provides a direct link between the tRNA-binding site and the movements in the head of the small subunit seen during translocation, thereby providing a possible pathway of signal transduction. We have created and characterized an rpsM(S13)-deficient strain of Escherichia coli and have found significant defects in subunit association, initiation and translocation through in vitro assays of S13-deficient ribosomes. Targeted mutagenesis of specific bridge and tRNA contact elements in S13 provides evidence that these two interaction domains play critical roles in maintaining the fidelity of translation. This ribosomal protein thus appears to play a non-essential, yet important role by modulating subunit interactions in multiple steps of the translation cycle.
核糖体蛋白S13位于小亚基的头部区域,在那里它通过其延伸的C末端与大核糖体亚基的中央突起以及与P位点结合的tRNA相互作用。大小亚基之间的桥接相互作用是动态的,并且被认为在协调翻译循环的分子运动中至关重要。S13在转位过程中在tRNA结合位点和小亚基头部的运动之间提供了直接联系,从而提供了一条可能的信号转导途径。我们构建并鉴定了一株大肠杆菌rpsM(S13)缺陷菌株,并通过对S13缺陷核糖体的体外测定发现了亚基结合、起始和转位方面的显著缺陷。对S13中特定桥接和tRNA接触元件的靶向诱变提供了证据,表明这两个相互作用结构域在维持翻译保真度方面起着关键作用。因此,这种核糖体蛋白似乎通过在翻译循环的多个步骤中调节亚基相互作用发挥着非必需但重要的作用。