Probing the kinetic and thermodynamic consequences of the tetraloop/tetraloop receptor monovalent ion-binding site in P4-P6 RNA by smFRET.
Probing the kinetic and thermodynamic consequences of the tetraloop/tetraloop receptor monovalent ion-binding site in P4-P6 RNA by smFRET.
复制标题
通过SMFRET探测四边形/四边形受体单价离子结合位点的动力学和热力学后果。
DOI：
10.1042/bst20140268
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发表时间：
2015-04
影响因子：
3.9
通讯作者：
Herschlag D
中科院分区：
文献类型：
--
作者：
Bisaria N;Herschlag D
结构化 RNA 分子在核心生物过程中发挥着重要作用，了解控制 RNA 折叠动力学和热力学的基本力和特征有助于阐明生物功能的原理。在这里，我们研究了这样一个特征，即单价阳离子与结构化 RNA（四膜虫核酶的 P4-P6 结构域）的特异性相互作用。我们采用单分子 FRET (smFRET) 方法，因为这些方法可以在广泛的稳定性范围内确定折叠平衡和速率常数，从而无需外推即可进行直接比较。这些实验为单价阳离子 Na+ 和 K+ 与 RNA 四环-四环受体 (TL-TLR) 三级基序的特异性结合提供了额外的证据。这些离子促进折叠和展开，与帮助命令 TLR 进行结合并在达到折叠过渡状态后进一步稳定三级接触的能力一致。
Structured RNA molecules play roles in central biological processes and understanding the basic forces and features that govern RNA folding kinetics and thermodynamics can help elucidate principles that underlie biological function. Here we investigate one such feature, the specific interaction of monovalent cations with a structured RNA, the P4–P6 domain of the Tetrahymena ribozyme. We employ single molecule FRET (smFRET) approaches as these allow determination of folding equilibrium and rate constants over a wide range of stabilities and thus allow direct comparisons without the need for extrapolation. These experiments provide additional evidence for specific binding of monovalent cations, Na+ and K+, to the RNA tetraloop–tetraloop receptor (TL–TLR) tertiary motif. These ions facilitate both folding and unfolding, consistent with an ability to help order the TLR for binding and further stabilize the tertiary contact subsequent to attainment of the folding transition state.