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Dynamics of Proofreading by the E. coli Pol III Replicase

Title
Dynamics of Proofreading by the E. coli Pol III Replicase
Authors
Jonghyun ParkSlobodan JergicYongmoon JeonWon-Ki ChoRyanggeun LeeNicholas E. DixonLEE, JONG BONG
POSTECH Authors
LEE, JONG BONG
Date Issued
Jan-2018
Publisher
Cell Press
Abstract
The αɛθ core of Escherichia coli DNA polymerase III (Pol III) associates with the β2 sliding clamp to processively synthesize DNA and remove misincorporated nucleotides. The α subunit is the polymerase while ɛ is the 3′ to 5′ proofreading exonuclease. In contrast to the polymerase activity of Pol III, dynamic features of proofreading are poorly understood. We used single-molecule assays to determine the excision rate and processivity of the β2-associated Pol III core, and observed that both properties are enhanced by mutational strengthening of the interaction between ɛ and β2. Thus, the ɛ-β2 contact is maintained in both the synthesis and proofreading modes. Remarkably, single-molecule real-time fluorescence imaging revealed the dynamics of transfer of primer-template DNA between the polymerase and proofreading sites, showing that it does not involve breaking of the physical interaction between ɛ and β2.
URI
http://oasis.postech.ac.kr/handle/2014.oak/38938
DOI
10.1016/j.chembiol.2017.09.008
ISSN
2451-9448
Article Type
Article
Citation
Cell Chemical Biology, 2018-01
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 LEE, JONG BONG
Dept of Physics
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