2026-04-29T04:00:30Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/121822024-11-29T21:13:14Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597
Repisalud
author
Dodd, Thomas
author
Botto, Margherita
author
Paul, Fabian
author
Lamers, Meindert H
author
Ivanov, Ivaylo
author
Fernandez-Leiro, Rafael
author
Dodd, Thomas
author
Botto, Margherita
author
Paul, Fabian
author
Lamers, Meindert H
author
Ivanov, Ivaylo
author
Fernandez-Leiro, Rafael
funder
United States Department of Health and Human Services
funder
National Science Foundation (Estados Unidos)
funder
United States Department of Energy
2021-03-10T09:31:48Z
2021-03-10T09:31:48Z
2020-11-01
Nat Commun.2020;11(1):5379.
http://hdl.handle.net/20.500.12105/12182
33097731
10.1038/s41467-020-19165-2
2041-1723
Nature communications
Proofreading by replicative DNA polymerases is a fundamental mechanism ensuring DNA replication fidelity. In proofreading, mis-incorporated nucleotides are excised through the 3'-5' exonuclease activity of the DNA polymerase holoenzyme. The exonuclease site is distal from the polymerization site, imposing stringent structural and kinetic requirements for efficient primer strand transfer. Yet, the molecular mechanism of this transfer is not known. Here we employ molecular simulations using recent cryo-EM structures and biochemical analyses to delineate an optimal free energy path connecting the polymerization and exonuclease states of E. coli replicative DNA polymerase Pol III. We identify structures for all intermediates, in which the transitioning primer strand is stabilized by conserved Pol III residues along the fingers, thumb and exonuclease domains. We demonstrate switching kinetics on a tens of milliseconds timescale and unveil a complete pol-to-exo switching mechanism, validated by targeted mutational experiments.
eng
Polymerization and editing modes of a high-fidelity DNA polymerase are linked by a well-defined path.
journal article
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