Analysis of the RNA Binding Specificity Landscape of C5 Protein Reveals Structure and Sequence Preferences that Direct RNase P Specificity.
Analysis of the RNA Binding Specificity Landscape of C5 Protein Reveals Structure and Sequence Preferences that Direct RNase P Specificity.
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DOI：
10.1016/j.chembiol.2016.09.002
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发表时间：
2016-10-20
影响因子：
8.6
通讯作者：
Harris ME
中科院分区：
文献类型：
--
作者：
Lin HC;Zhao J;Niland CN;Tran B;Jankowsky E;Harris ME
RNA结合蛋白（RBP）通常参与非平衡细胞过程，并且特异性可以由替代RNA的结合反应的基态、过渡态或产物态的差异引起。在这里，我们使用高通量的方法来测量和分析所有可能的序列组合的前体tRNA结合位点的C5，大肠杆菌核糖核酸酶P的必需蛋白亚基的RNA缔合动力学和平衡结合亲和力。结果表明，C5的RNA序列特异性的出现是由于有利的RNA-蛋白质相互作用，稳定的过渡状态的协会和绑定ES复合物。特异性进一步受到涉及基态C5结合位点的不利RNA结构的影响。结果说明了一个全面的定量分析方法的RNA结合特异性，并显示如何RNA的结构和序列偏好的一个必需的蛋白质亚基直接的特异性的核糖核蛋白酶。Lin等使用高通量方法分析细菌核糖核酸酶P蛋白结合位点中所有可能的序列组合的结合动力学和平衡亲和力。结果揭示了RNA结构和序列如何指导核糖核蛋白酶的特异性。
RNA-binding proteins (RBPs) are typically involved in non-equilibrium cellular processes, and specificity can arise from differences in ground state, transition state or product states of the binding reactions for alternative RNAs. Here, we use high throughput methods to measure and analyze the RNA association kinetics and equilibrium binding affinity for all possible sequence combinations in precursor tRNA binding site of C5, the essential protein subunit of Escherichia coli ribonuclease P. The results show that the RNA sequence specificity of C5 arises due to favorable RNA-protein interactions that stabilize the transition state for association and bound ES complex. Specificity is further impacted by unfavorable RNA structure involving the C5 binding site in the ground state. The results illustrate a comprehensive quantitative approach for analysis of RNA binding specificity, and show how both RNA structure and sequence preferences of an essential protein subunit direct the specificity of a ribonucleoprotein enzyme. Lin et al. use high throughput methods to analyze the binding kinetics and equilibrium affinities for all possible sequence combinations in binding site of bacterial ribonuclease P protein. The results reveal how both RNA structure and sequence direct the specificity of a ribonucleoprotein enzyme.
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影响因子：
2.9
作者：
Crary, SM;Niranjanakumari, S;Fierke, CA
通讯作者：
Fierke, CA
DOI：
10.1073/pnas.90.16.7814
发表时间：
1993-08-15
影响因子：
11.1
作者：
MATOUSCHEK, A;FERSHT, AR
通讯作者：
FERSHT, AR
影响因子：
16.6
作者：
Lassila JK;Zalatan JG;Herschlag D
通讯作者：
Herschlag D
影响因子：
2.9
作者：
Kurz, JC;Niranjanakumari, S;Fierke, CA
通讯作者：
Fierke, CA
DOI：
10.1186/1748-7188-6-26
发表时间：
2011-11-24
期刊：
Algorithms for molecular biology : AMB
影响因子：
--
作者：
Lorenz R;Bernhart SH;Höner Zu Siederdissen C;Tafer H;Flamm C;Stadler PF;Hofacker IL
通讯作者：
Hofacker IL