期刊
NATURE CHEMICAL BIOLOGY
卷 15, 期 5, 页码 472-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41589-019-0267-9
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资金
- DOE Office of Science [DE-AC02-06CH11357]
- National Institute of General Medical Sciences from the National Institutes of Health [P30 GM124165]
- NIH-ORIP HEI grant [S10 RR029205]
- National Institutes of Health [S10_RR25528, S10_RR028976]
- LabEx NetRNA [ANR-10-LABX-0036]
- NSERC (Canada)
- Agence Nationale pour la Recherche [ANR-16-CE11-0010-01]
- Intramural Program of the NHLBI, NIH
- Agence Nationale de la Recherche (ANR) [ANR-16-CE11-0010] Funding Source: Agence Nationale de la Recherche (ANR)
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [ZIAHL001452, ZIAHL006188] Funding Source: NIH RePORTER
Several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the similar to 30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A1OU), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans Watson-Crick pair (whose A1OU mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A1OU) fluoresce similar to 50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.
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