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Elife. 2017 Dec 6;6. pii: e32177. doi: 10.7554/eLife.32177.

Single-molecule studies contrast ordered DNA replication with stochastic translesion synthesis.

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MRC laboratory of Molecular Biology, Cambridge, United Kingdom.


High fidelity replicative DNA polymerases are unable to synthesize past DNA adducts that result from diverse chemicals, reactive oxygen species or UV light. To bypass these replication blocks, cells utilize specialized translesion DNA polymerases that are intrinsically error prone and associated with mutagenesis, drug resistance, and cancer. How untimely access of translesion polymerases to DNA is prevented is poorly understood. Here we use co-localization single-molecule spectroscopy (CoSMoS) to follow the exchange of the E. coli replicative DNA polymerase Pol IIIcore with the translesion polymerases Pol II and Pol IV. We find that in contrast to the toolbelt model, the replicative and translesion polymerases do not form a stable complex on one clamp but alternate their binding. Furthermore, while the loading of clamp and Pol IIIcore is highly organized, the exchange with the translesion polymerases is stochastic and is not determined by lesion-recognition but instead a concentration-dependent competition between the polymerases.


CoSMoS; DNA replication; E. coli; biochemistry; biophysics; single-molecule; structural biology; translesion DNA synthesis

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