Structure-based investigation of fluorogenic Pepper aptamer
Structure-based investigation of fluorogenic Pepper aptamer
复制标题
基于结构的荧光辣椒适体研究
DOI：
10.1038/s41589-021-00884-6
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发表时间：
2021-11-01
影响因子：
14.8
通讯作者：
Ren, Aiming
中科院分区：
文献类型：
--
作者：
Huang, Kaiyi;Chen, Xianjun;Ren, Aiming
Structural analysis of the Pepper aptamer in complex with its cognate HBC or HBC-like color variants reveals that it binds fluorophore molecules via one non-G-quadruplex base quadruple and one noncanonical G center dot U base pair.Pepper fluorescent RNAs are a recently reported bright, stable and multicolor fluorogenic aptamer tag that enable imaging of diverse RNAs in live cells. To investigate the molecular basis of the superior properties of Pepper, we determined the structures of complexes of Pepper aptamer bound with its cognate HBC or HBC-like fluorophores at high resolution by X-ray crystallography. The Pepper aptamer folds in a monomeric non-G-quadruplex tuning-fork-like architecture composed of a helix and one protruded junction region. The near-planar fluorophore molecule intercalates in the middle of the structure and is sandwiched between one non-G-quadruplex base quadruple and one noncanonical G center dot U wobble helical base pair. In addition, structure-based mutational analysis is evaluated by in vitro and live-cell fluorogenic detection. Taken together, our research provides a structural basis for demystifying the fluorescence activation mechanism of Pepper aptamer and for further improvement of its future application in RNA visualization.
