Ligand-modulated folding of the full-length adenine riboswitch probed by NMR and single-molecule FRET spectroscopy.
Ligand-modulated folding of the full-length adenine riboswitch probed by NMR and single-molecule FRET spectroscopy.
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DOI：
10.1093/nar/gkx110
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发表时间：
2017-05-19
影响因子：
14.9
通讯作者：
Schwalbe H
中科院分区：
文献类型：
--
作者：
Warhaut S;Mertinkus KR;Höllthaler P;Fürtig B;Heilemann M;Hengesbach M;Schwalbe H
创伤弧菌的全长翻译调节加腺嘌呤核糖开关(ASW)比其分离的适配子结构域具有更复杂的构象空间。除了预测的apo(ApoA)和holo构象具有保守的三向连接嘌呤核糖开关适配子外，它还采用第二apo(ApoB)构象，具有完全不同的二级结构。在这里，我们用核磁共振和单分子FRET(SmFRET)光谱表征了全长ADD ASW的配体依赖的构象动力学。这两种方法都揭示了腺嘌呤诱导的二级结构从apoB形式到apoA形式的转换，不涉及适配子和表达平台之间的三级结构相互作用。这有力地表明，ADD ASW通过捕获全息态的apoA-型二级结构来触发翻译。有趣的是，核磁共振显示apoA和holo有一个均质的、对接的适配子亲吻环折叠，而smFRET则显示了apoA和holo构象的相对稳定、未对接和对接的亚态之间持续的适配子亲吻环对接动力学。因此，解开大的连接核糖开关的折叠需要整合互补的溶液结构技术，如核磁共振和smFRET。
The full-length translation-regulating add adenine riboswitch (Asw) from Vibrio vulnificus has a more complex conformational space than its isolated aptamer domain. In addition to the predicted apo (apoA) and holo conformation that feature the conserved three-way junctional purine riboswitch aptamer, it adopts a second apo (apoB) conformation with a fundamentally different secondary structure. Here, we characterized the ligand-dependent conformational dynamics of the full-length add Asw by NMR and by single-molecule FRET (smFRET) spectroscopy. Both methods revealed an adenine-induced secondary structure switch from the apoB-form to the apoA-form that involves no tertiary structural interactions between aptamer and expression platform. This strongly suggests that the add Asw triggers translation by capturing the apoA-form secondary structure in the holo state. Intriguingly, NMR indicated a homogenous, docked aptamer kissing loop fold for apoA and holo, while smFRET showed persistent aptamer kissing loop docking dynamics between comparably stable, undocked and docked substates of the apoA and the holo conformation. Unraveling the folding of large junctional riboswitches thus requires the integration of complementary solution structural techniques such as NMR and smFRET.
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影响因子：
4.1
作者：
Nozinovic S;Reining A;Kim YB;Noeske J;Schlepckow K;Wöhnert J;Schwalbe H
通讯作者：
Schwalbe H
腺嘌呤感应核糖开关适体结构域的金属离子结合和金属离子诱导折叠。
影响因子：
14.9
作者：
Noeske J;Schwalbe H;Wöhnert J
通讯作者：
Wöhnert J
影响因子：
16.6
作者：
Hengesbach, Martin;Kim, Nak-Kyoon;Feigon, Juli;Stone, Michael D.
通讯作者：
Stone, Michael D.
影响因子：
64.8
作者：
Reining, Anke;Nozinovic, Senada;Schwalbe, Harald
通讯作者：
Schwalbe, Harald
影响因子：
14.8
作者：
Haller, Andrea;Rieder, Ulrike;Micura, Ronald
通讯作者：
Micura, Ronald