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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Comprehensive genetic selection revealed essential bases in the peptidyl-transferase center
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Comprehensive genetic selection revealed essential bases in the peptidyl-transferase center

机译：全面的遗传选择揭示了肽基转移酶中心的重要碱基

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During protein synthesis, the ribosome catalyzes peptide-bond formation. Biochemical and structural studies revealed that conserved nucleoticles in the pepticlyl-transferase center (PTC) and its proximity may play a key role in pepticle-bond formation; the exact mechanism involved remains unclear. To more precisely define the functional importance of the highly conserved residues, we used a systematic genetic method, which we named SSER (systematic selection of functional sequences by enforced replacement), that allowed us to identify essential nucleoticles for ribosomal function from randomized rRNA libraries in Escherichia coli cells. These libraries were constructed by complete randomization of the critical regions in and around the FTC. The selected variants contained natural rRNA sequences from other organisms and organelles as well as unnatural functional sequences; hence providing insights into the functional roles played by these essential bases and suggesting how the universal catalytic mechanism of pepticle-bond formation could evolve in all living organisms. Our results highlight essential bases and interactions, which are shaping the PTC architecture and guiding the motions of the tRNA terminus from the A to the P site, found to be crucial not only for the formation of the pepticle bond but also for nascent chain elongation.
机译：在蛋白质合成过程中，核糖体催化肽键的形成。生化和结构研究表明，胃蛋白酶转移酶中心（PTC）及其附近的保守核苷酸可能在胃蛋白酶键的形成中起关键作用。确切的机制尚不清楚。为了更精确地定义高度保守残基的功能重要性，我们使用了一种系统遗传方法，我们将其命名为SSER（通过强制置换对功能序列进行系统选择），这使我们能够从随机rRNA文库中鉴定核糖体功能的基本核苷酸。大肠杆菌细胞。这些库是通过将FTC及其周围的关键区域完全随机化而构建的。选择的变体包含来自其他生物和细胞器的天然rRNA序列以及非天然功能序列。因此，提供了对这些必不可少的基础所发挥的功能性作用的见解，并提出了在所有活生物体中消化键形成的普遍催化机制将如何进化。我们的研究结果突出了必不可少的基础和相互作用，这些基础和相互作用正在塑造PTC的体系结构并指导tRNA末端从A到P位点的运动，这不仅对形成消化键而且对新生链的延长至关重要。

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