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Cryo-EM structure of Pol κ−DNA−PCNA holoenzyme and implications for polymerase switching in DNA lesion bypass

View ORCID ProfileClaudia Lancey, View ORCID ProfileMuhammad Tehseen, View ORCID ProfileMasateru Takahashi, Mohamed A. Sobhy, View ORCID ProfileTimothy J. Ragan, View ORCID ProfileRamon Crehuet, View ORCID ProfileSamir M. Hamdan, View ORCID ProfileAlfredo De Biasio
doi: https://doi.org/10.1101/2020.07.10.196956
Claudia Lancey
1Leicester Institute of Structural & Chemical Biology and Department of Molecular & Cell Biology, University of Leicester, Lancaster Rd, Leicester LE1 7HB, UK
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Muhammad Tehseen
2Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal 23955, Saudi Arabia
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Masateru Takahashi
2Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal 23955, Saudi Arabia
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Mohamed A. Sobhy
2Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal 23955, Saudi Arabia
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Timothy J. Ragan
1Leicester Institute of Structural & Chemical Biology and Department of Molecular & Cell Biology, University of Leicester, Lancaster Rd, Leicester LE1 7HB, UK
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Ramon Crehuet
3CSIC-Institute for Advanced Chemistry of Catalonia (IQAC) C/ Jordi Girona 18-26, 08034 Barcelona, Spain
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Samir M. Hamdan
2Division of Biological and Environmental Sciences and Engineering, King Abdullah University of Science and Technology, Thuwal 23955, Saudi Arabia
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  • For correspondence: samir.hamdan@kaust.edu.sa adb43@leicester.ac.uk
Alfredo De Biasio
1Leicester Institute of Structural & Chemical Biology and Department of Molecular & Cell Biology, University of Leicester, Lancaster Rd, Leicester LE1 7HB, UK
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  • For correspondence: samir.hamdan@kaust.edu.sa adb43@leicester.ac.uk
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Abstract

Replacement of the stalled replicative polymerase (Pol δ) at a DNA lesion by the error-prone DNA polymerase κ (Pol κ) restarts synthesis past the lesion to prevent genome instability. The switching from Pol δ to Pol κ is mediated by the processivity clamp PCNA but the structural basis of this mechanism is unknown. We determined the Cryo-EM structures of human Pol κ–DNA–PCNA complex and of a stalled Pol δ–DNA–PCNA complex at 3.9 and 4.7 Å resolution, respectively. In Pol κ complex, the C-terminus of the PAD domain docks the catalytic core to one PCNA protomer in an angled orientation, bending the DNA exiting Pol κ active site through PCNA. In Pol δ complex, the DNA is disengaged from the active site but is retained by the thumb domain. We present a model for polymerase switching facilitated by Pol κ recruitment to PCNA and Pol κ conformational sampling to seize the DNA from stalled Pol δ assisted by PCNA tilting.

Competing Interest Statement

The authors have declared no competing interest.

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Posted July 10, 2020.
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Cryo-EM structure of Pol κ−DNA−PCNA holoenzyme and implications for polymerase switching in DNA lesion bypass
Claudia Lancey, Muhammad Tehseen, Masateru Takahashi, Mohamed A. Sobhy, Timothy J. Ragan, Ramon Crehuet, Samir M. Hamdan, Alfredo De Biasio
bioRxiv 2020.07.10.196956; doi: https://doi.org/10.1101/2020.07.10.196956
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Cryo-EM structure of Pol κ−DNA−PCNA holoenzyme and implications for polymerase switching in DNA lesion bypass
Claudia Lancey, Muhammad Tehseen, Masateru Takahashi, Mohamed A. Sobhy, Timothy J. Ragan, Ramon Crehuet, Samir M. Hamdan, Alfredo De Biasio
bioRxiv 2020.07.10.196956; doi: https://doi.org/10.1101/2020.07.10.196956

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