Imperial College London
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Professor David S. Rueda

Faculty of Medicine, Department of Infectious Disease

Chair in Molecular and Cellular Biophysics
 
 
 
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Contact

 

david.rueda Website

 
 
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Location

 

6.12DLMS BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Gahlon:2018:10.1039/C8CP04112A,
author = {Gahlon, HL and Walker, AR and Cisneros, GA and Lamers, MH and Rueda, DS},
doi = {10.1039/C8CP04112A},
journal = {Physical Chemistry Chemical Physics},
pages = {26892--26902},
title = {Reduced structural flexibility for an exonuclease deficient DNA polymerase III mutant},
url = {http://dx.doi.org/10.1039/C8CP04112A},
volume = {20},
year = {2018}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - DNA synthesis, carried out by DNA polymerases, requires balancing speed and accuracy for faithful replication of the genome. High fidelity DNA polymerases contain a 3′–5′ exonuclease domain that can remove misincorporated nucleotides on the 3′ end of the primer strand, a process called proofreading. The E. coli replicative polymerase, DNA polymerase III, has spatially separated (∼55 Å apart) polymerase and exonuclease subunits. Here, we report on the dynamics of E. coli DNA polymerase III proofreading in the presence of its processivity factor, the β2-sliding clamp, at varying base pair termini using single-molecule FRET. We find that the binding kinetics do not depend on the base identity at the termini, indicating a tolerance for DNA mismatches. Further, our single-molecule data and MD simulations show two previously unobserved features: (1) DNA Polymerase III is a highly dynamic protein that adopts multiple conformational states while bound to DNA with matched or mismatched ends, and (2) an exonuclease-deficient DNA polymerase III has reduced conformational flexibility. Overall, our single-molecule experiments provide high time-resolution insight into a mechanism that ensures high fidelity DNA replication to maintain genome integrity.
AU  - Gahlon,HL
AU  - Walker,AR
AU  - Cisneros,GA
AU  - Lamers,MH
AU  - Rueda,DS
DO  - 10.1039/C8CP04112A
EP  - 26902
PY  - 2018///
SN  - 1463-9076
SP  - 26892
TI  - Reduced structural flexibility for an exonuclease deficient DNA polymerase III mutant
T2  - Physical Chemistry Chemical Physics
UR  - http://dx.doi.org/10.1039/C8CP04112A
UR  - http://hdl.handle.net/10044/1/64099
VL  - 20
ER  -
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