%0 Journal Article
%A Borggräfe, Jan
%A Victor, Julian
%A Rosenbach, Hannah
%A Viegas, Aldino
%A Gertzen, Christoph G. W.
%A Wuebben, Christine
%A Kovacs, Helena
%A Gopalswamy, Mohanraj
%A Riesner, Detlev
%A Steger, Gerhard
%A Schiemann, Olav
%A Gohlke, Holger
%A Span, Ingrid
%A Etzkorn, Manuel
%T Time-resolved structural analysis of an RNA-cleaving DNA catalyst
%J Nature 
%V 601
%@ 0028-0836
%C London [u.a.]
%I Nature Publ. Group
%M FZJ-2021-06169
%P 144-149
%D 2022
%X The 10–23 DNAzyme is one of the most prominent catalytically active DNA sequences1,2. Its ability to cleave a wide range of RNA targets with high selectivity entails a substantial therapeutic and biotechnological potential2. However, the high expectations have not yet been met, a fact that coincides with the lack of high-resolution and time-resolved information about its mode of action3. Here we provide high-resolution NMR characterization of all apparent states of the prototypic 10–23 DNAzyme and present a comprehensive survey of the kinetics and dynamics of its catalytic function. The determined structure and identified metal-ion-binding sites of the precatalytic DNAzyme–RNA complex reveal that the basis of the DNA-mediated catalysis is an interplay among three factors: an unexpected, yet exciting molecular architecture; distinct conformational plasticity; and dynamic modulation by metal ions. We further identify previously hidden rate-limiting transient intermediate states in the DNA-mediated catalytic process via real-time NMR measurements. Using a rationally selected single-atom replacement, we could considerably enhance the performance of the DNAzyme, demonstrating that the acquired knowledge of the molecular structure, its plasticity and the occurrence of long-lived intermediate states constitutes a valuable starting point for the rational design of next-generation DNAzymes.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ pmid:34949858
%U <Go to ISI:>//WOS:000734154500001
%R 10.1038/s41586-021-04225-4
%U https://juser.fz-juelich.de/record/904599