Factors that influence T box riboswitch efficacy and tRNA affinity.
Factors that influence T box riboswitch efficacy and tRNA affinity.
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影响 T 盒核糖开关功效和 tRNA 亲和力的因素。
DOI：
10.1016/j.bmc.2015.07.018
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发表时间：
2015
影响因子：
3.5
通讯作者：
Hines,JV
中科院分区：
文献类型：
--
作者：
Zeng,C;Zhou,S;Bergmeier,SC;Hines,JV
T盒核糖开关是一个有趣的潜在的抗菌药物发现的目标。核糖开关主要存在于革兰氏阳性细菌中，通过选择性地响应不带电荷的tRNA来控制转录通读，从而调节基因表达。已知多胺和分子拥挤特异性地影响RNA功能，但它们对T盒核糖开关功效和tRNA亲和力的影响尚未完全表征。开发了荧光监测的体外转录测定法，以容易地量化这些分子相互作用，并为全面的药物发现筛选级联提供中等通量的功能测定。多胺亚精胺特异性增强T盒核糖开关通读功效，EC50= 0.58 mM，不依赖于tRNA结合。通过加入聚乙二醇来模拟分子拥挤，对tRNA对核糖开关的亲和力没有影响，但确实降低了tRNA诱导的通读的功效。这些结果表明，T盒核糖开关tRNA亲和力和通读功效是由环境因素错综复杂地调制。
The T box riboswitch is an intriguing potential target for antibacterial drug discovery. Found primarily in Gram-positive bacteria, the riboswitch regulates gene expression by selectively responding to uncharged tRNA to control transcription readthrough. Polyamines and molecular crowding are known to specifically affect RNA function, but their effect on T box riboswitch efficacy and tRNA affinity have not been fully characterized. A fluorescence-monitored in vitro transcription assay was developed to readily quantify these molecular interactions and to provide a moderate-throughput functional assay for a comprehensive drug discovery screening cascade. The polyamine spermidine specifically enhanced T box riboswitch readthrough efficacy with an EC50= 0.58 mM independent of tRNA binding. Molecular crowding, simulated by the addition of polyethylene glycol, had no effect on tRNA affinity for the riboswitch, but did reduce the efficacy of tRNA-induced readthrough. These results indicate that the T box riboswitch tRNA affinity and readthrough efficacy are intricately modulated by environmental factors.
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影响因子：
18.3
作者：
Deigan, Katherine E.;Ferre-D'Amare, Adrian R.
通讯作者：
Ferre-D'Amare, Adrian R.
结合两个 tRNA 的双密码子 T 盒核糖开关
DOI：
--
发表时间：
2013
影响因子：
11.1
作者：
Nizar Y. Saad;Vassiliki Stamatopoulou;Mélanie Brayé;D. Drainas;C. Stathopoulos;H. Becker
通讯作者：
H. Becker
影响因子：
5.6
作者：
Yousef, MR;Grundy, FJ;Henkin, TM
通讯作者：
Henkin, TM
影响因子：
7.3
作者：
Orac, Crina M.;Zhou, Shu;Means, John A.;Boehm, David;Bergmeier, Stephen C.;Hines, Jennifer V.
通讯作者：
Hines, Jennifer V.
影响因子：
7.4
作者：
Lee, Janghyun;Vogt, Catherine E.;McBrairty, Mitchell;Al-Hashimi, Hashim M.
通讯作者：
Al-Hashimi, Hashim M.