Structure and functional reselection of the Mango-III fluorogenic RNA aptamer

Trachman III, Robert J.; Autour, Alexis; Jeng, Sunny C.  Y.; Abdolahzadeh, Amir; Andreoni, Alessio; Cojocaru, Razvan; Garipov, Ramil; Dolgosheina, Elena V.; Knutson, Jay R.; Ryckelynck, Michael; Unrau, Peter J.; Ferré-D’Amaré, Adrian R.

doi:10.1038/s41589-019-0267-9

Structure and functional reselection of the Mango-III fluorogenic RNA aptamer

Journal Article · Mon Apr 15 00:00:00 EDT 2019 · Nature Chemical Biology

DOI:https://doi.org/10.1038/s41589-019-0267-9· OSTI ID:1512999

Trachman III, Robert J. ^[1]; Autour, Alexis ^[2]; Jeng, Sunny C. Y. ^[3]; Abdolahzadeh, Amir ^[3]; ^[1]; Cojocaru, Razvan ^[3]; Garipov, Ramil ^[3]; Dolgosheina, Elena V. ^[3]; Knutson, Jay R. ^[1]; ^[2]; Unrau, Peter J. ^[3]; ^[1]

National Heart, Lung, and Blood Inst., Bethesda, MD (United States)
Univ. de Strasbourg (France)
Simon Fraser Univ., Burnaby, BC (Canada)

Here, several turn-on RNA aptamers that activate small-molecule fluorophores have been selected in vitro. Among these, the ~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(A10U), 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 A10U 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(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); Natural Sciences and Engineering Research Council of Canada (NSERC); LabEx NetRNA; Agence Nationale de la Recherché (ANR); National Heart, Lung and Blood Institute (NHLBI)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1512999

Journal Information:: Nature Chemical Biology, Journal Name: Nature Chemical Biology Journal Issue: 5 Vol. 15; ISSN 1552-4450

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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