TY  - JOUR
AU  - Borggräfe, Jan
AU  - Victor, Julian
AU  - Rosenbach, Hannah
AU  - Viegas, Aldino
AU  - Gertzen, Christoph G. W.
AU  - Wuebben, Christine
AU  - Kovacs, Helena
AU  - Gopalswamy, Mohanraj
AU  - Riesner, Detlev
AU  - Steger, Gerhard
AU  - Schiemann, Olav
AU  - Gohlke, Holger
AU  - Span, Ingrid
AU  - Etzkorn, Manuel
TI  - Time-resolved structural analysis of an RNA-cleaving DNA catalyst
JO  - Nature 
VL  - 601
SN  - 0028-0836
CY  - London [u.a.]
PB  - Nature Publ. Group
M1  - FZJ-2021-06169
SP  - 144-149
PY  - 2022
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - pmid:34949858
UR  - <Go to ISI:>//WOS:000734154500001
DO  - DOI:10.1038/s41586-021-04225-4
UR  - https://juser.fz-juelich.de/record/904599
ER  -