Fluorophore ligand binding and complex stabilization of the RNA Mango and RNA Spinach aptamers.
Fluorophore ligand binding and complex stabilization of the RNA Mango and RNA Spinach aptamers.
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DOI：
10.1261/rna.056226.116
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发表时间：
2016-12
期刊：
影响因子：
--
通讯作者：
Unrau PJ
中科院分区：
文献类型：
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作者：
Jeng SC;Chan HH;Booy EP;McKenna SA;Unrau PJ
生物RNA和RNA蛋白质复合物的有效追踪和纯化目前具有挑战性。同时解决这两个问题的一个有希望的策略是开发针对可标记小分子荧光团的高亲和力RNA适体。RNA Mango是一种39个核苷酸的平行链G-四链体RNA适体基序，其以纳摩尔亲和力与一组噻唑橙子（TO 1）衍生物结合，同时诱导荧光增加103倍。我们发现，RNA芒果有一个很大的增加，其热稳定性后，加入其TO 1-生物素配体。与这种热稳定性相一致，RNA Mango可以有效地将TO 1-生物素与广泛的小分子荧光团区分开来。相比之下，RNA菠菜，这是已知的具有实质上更刚性的G-四链体结构，被发现结合到这组荧光团，往往具有更高的亲和力比其天然配体，3，5-二氟-4-羟基苯亚甲基咪唑啉酮（DFHBI），并没有表现出热稳定的TO 1-生物素荧光团的存在下。我们的数据表明，RNA Mango可能使用一种协调的配体结合机制，使其能够同时结合和识别其TO 1-生物素配体，而RNA Spinach似乎缺乏这种机制。RNA Mango的高结合亲和力和荧光效率为RNA Spinach作为RNA报告系统提供了一种引人注目的替代方案，并为未来开发小荧光团RNA报告系统铺平了道路。
The effective tracking and purification of biological RNAs and RNA protein complexes is currently challenging. One promising strategy to simultaneously address both of these problems is to develop high-affinity RNA aptamers against taggable small molecule fluorophores. RNA Mango is a 39-nucleotide, parallel-stranded G-quadruplex RNA aptamer motif that binds with nanomolar affinity to a set of thiazole orange (TO1) derivatives while simultaneously inducing a 103-fold increase in fluorescence. We find that RNA Mango has a large increase in its thermal stability upon the addition of its TO1-Biotin ligand. Consistent with this thermal stabilization, RNA Mango can effectively discriminate TO1-Biotin from a broad range of small molecule fluorophores. In contrast, RNA Spinach, which is known to have a substantially more rigid G-quadruplex structure, was found to bind to this set of fluorophores, often with higher affinity than to its native ligand, 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI), and did not exhibit thermal stabilization in the presence of the TO1-Biotin fluorophore. Our data suggest that RNA Mango is likely to use a concerted ligand-binding mechanism that allows it to simultaneously bind and recognize its TO1-Biotin ligand, whereas RNA Spinach appears to lack such a mechanism. The high binding affinity and fluorescent efficiency of RNA Mango provides a compelling alternative to RNA Spinach as an RNA reporter system and paves the way for the future development of small fluorophore RNA reporter systems.
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影响因子：
14.9
作者：
Renaud de la Faverie A;Guédin A;Bedrat A;Yatsunyk LA;Mergny JL
通讯作者：
Mergny JL
影响因子：
3.7
作者：
Ariyo, Emmanuel O.;Booy, Evan P.;McKenna, Sean A.
通讯作者：
McKenna, Sean A.
DOI：
10.1126/science.1207339
发表时间：
2011-07-29
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
Paige JS;Wu KY;Jaffrey SR
通讯作者：
Jaffrey SR
影响因子：
14.9
作者：
Autour A;Westhof E;Ryckelynck M
通讯作者：
Ryckelynck M
DOI：
10.1038/nrm3918
发表时间：
2015-02
期刊：
Nature reviews. Molecular cell biology
影响因子：
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作者：
通讯作者：
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