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Capture and Release of tRNA by the T-Loop Receptor in the Function of the T-Box Riboswitch

机译:T型环接收器在T型盒子riboswitch的功能中捕获和释放TRNA

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摘要

In Gram-positive bacteria, the tRNA-dependent T-box riboswitch system regulates expression of amino acid biosynthetic and aminoacyl-tRNA synthetase genes through a transcription attenuation mechanism. Binding of uncharged tRNA "closes" the switch, allowing transcription read -through. Structural studies of the 100-nucleotide stem I domain reveal tRNA utilizes base pairing and stacking interactions to bind the stem, but little is known structurally about the 180-nucleotide riboswitch core (stem I, stem III, and antiterminator stem) in complex with tRNA or the mechanism of coupling of the intermolecular binding domains crucial to T-box function. Here we utilize solution structural and biophysical methods to characterize the interplay of the different riboswitch tRNA contact points using Bacillus subtilis and Oceanobacillus iheyensis glycyl T-box and T-box:tRNA constructs. The data reveal that tRNA:riboswitch core binding at equilibrium involves only Specifier anticodon and antiterminator acceptor stem pairing. The elbow:platform stacking interaction observed in studies of the T-box stem I domain is released after pairing between the acceptor stem and the bulge in the antiterminator helix. The results are consistent with the model of T-box riboswitch:tRNA function in which tRNA is captured by stem I of the nascent mRNA followed by stabilization of the antiterminator helix and the paused transcription complex.
机译:在革兰氏阳性细菌中,TRNA依赖性T字幕核糖系统通过转录衰减机制调节氨基酸生物合成和氨基酰基-CRNA合成酶基因的表达。无需特性的结合“关闭”开关,允许转录读取。 100核苷酸茎I结构域的结构研究揭示了TRNA利用碱基配对和堆叠相互作用来结合茎,但是在结构上围绕180核苷酸核糖核心(茎I,茎III和抗渗透剂茎)以复合物在结构上众所周知或者分子结合域的偶联机制至关重要的T字幕功能。在这里,我们利用解决方案结构和生物物理方法来表征使用枯草芽孢杆菌和海洋植物糖尿病T-Box和T盒:TRNA构建体的不同Riboswitch TRNA接触点的相互作用。数据显示,TRNA:核核心核心结合在平衡下涉及仅说明书抗ocodon和抗拒受体受体干法配对。肘部:在接收器杆和抗烧结器螺旋中的凸起之间配对后,在T字幕杆I结构域的研究中观察到的平台堆叠相互作用。结果与T型盒子核糖开关的模型一致:TRNA功能,其中TRNA被起始的mRNA的茎I捕获,然后稳定抗铝蛋白螺旋和暂停的转录复合物。

著录项

  • 来源
    《Biochemistry》 |2017年第28期|共10页
  • 作者单位

    Tsinghua Univ Sch Life Sci Beijing Adv Innovat Ctr Struct Biol Beijing 100084 Peoples R China;

    Rice Univ Dept Biosci Houston TX 77005 USA;

    Tsinghua Univ Sch Life Sci Beijing Adv Innovat Ctr Struct Biol Beijing 100084 Peoples R China;

    NCI Struct Biophys Lab Frederick MD 21701 USA;

    Rice Univ Dept Biosci Houston TX 77005 USA;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
  • 关键词

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