Crucial Roles of Two Hydrated Mg2+ Ions in Reaction Catalysis of the Pistol Ribozyme

Teplova, Marianna; Falschlunger, Christoph; Krasheninina, Olga; Egger, Michaela; Ren, Aiming; Patel, Dinshaw J.; Micura, Ronald

doi:10.1002/anie.201912522

Title: Crucial Roles of Two Hydrated Mg²⁺ Ions in Reaction Catalysis of the Pistol Ribozyme

Journal Article · 04 December 2019 · Angewandte Chemie (International Edition)

DOI: https://doi.org/10.1002/anie.201912522 · OSTI ID:1598035

Teplova, Marianna ^[1]; Falschlunger, Christoph ^[2]; Krasheninina, Olga ^[2]; Egger, Michaela ^[2]; Ren, Aiming ^[3]; Patel, Dinshaw J. ^[1];

^[2]

Memorial Sloan-Kettering Cancer Center, New York, NY (United States)
Leopold-Franzens Univ., Innsbruck (Austria)
Zhejiang Univ., Hangzhou (China)

Pistol ribozymes constitute a new class of small self-cleaving RNAs. Crystal structures have been solved, providing three-dimensional snapshots along the reaction coordinate of pistol phosphodiester cleavage, corresponding to the pre-catalytic state, a vanadate mimic of the transition state, and the product. The results led to the proposed underlying chemical mechanism. Importantly, a hydrated Mg²⁺ ion remains innersphere-coordinated to N7 of G33 in all three states, and is consistent with its likely role as acid in general acid base catalysis (δ and β catalysis). Strikingly, the new structures shed light on a second hydrated Mg²⁺ ion that approaches the scissile phosphate from its binding site in the pre-cleavage state to reach out for water-mediated hydrogen bonding in the cyclophosphate product. The major role of the second Mg²⁺ ion appears to be the stabilization of product conformation. This study delivers a mechanistic understanding of ribozyme-catalyzed backbone cleavage.

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

Sponsoring Organization:: Austrian Science Fund (FWF); National Institutes of Health (NIH); National Natural Science Foundation of China (NSFC); The Austrian Research Promotion Agency (FFG)

OSTI ID:: 1598035

Journal Information:: Angewandte Chemie (International Edition), Journal Name: Angewandte Chemie (International Edition) Journal Issue: 7 Vol. 59; ISSN 1433-7851

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Title: Crucial Roles of Two Hydrated Mg2+ Ions in Reaction Catalysis of the Pistol Ribozyme

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Title: Crucial Roles of Two Hydrated Mg²⁺ Ions in Reaction Catalysis of the Pistol Ribozyme