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Structure of the processive human Pol δ holoenzyme

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journal contribution
posted on 22.05.2020, 11:16 by C Lancey, M Tehseen, VS Raducanu, F Rashid, N Merino, TJ Ragan, CG Savva, MS Zaher, A Shirbini, FJ Blanco, SM Hamdan, A De Biasio
In eukaryotes, DNA polymerase δ (Pol δ) bound to the proliferating cell nuclear antigen (PCNA) replicates the lagging strand and cooperates with flap endonuclease 1 (FEN1) to process the Okazaki fragments for their ligation. We present the high-resolution cryo-EM structure of the human processive Pol δ–DNA–PCNA complex in the absence and presence of FEN1. Pol δ is anchored to one of the three PCNA monomers through the C-terminal domain of the catalytic subunit. The catalytic core sits on top of PCNA in an open configuration while the regulatory subunits project laterally. This arrangement allows PCNA to thread and stabilize the DNA exiting the catalytic cleft and recruit FEN1 to one unoccupied monomer in a toolbelt fashion. Alternative holoenzyme conformations reveal important functional interactions that maintain PCNA orientation during synthesis. This work sheds light on the structural basis of Pol δ’s activity in replicating the human genome.

Funding

This research was supported by King Abdullah University of Science and Technology through core funding (to S.M.H.), and by the Wellcome Trust (to A.D.B.). F.J.B. was supported by grant CTQ2017-83810-R from MCIU/AEI/FEDER, UE. CIC bioGUNE acknowledges MINECO for the Severo Ochoa Excellence Accreditation SEV-2016-0644.We acknowledge the Midlands Regional Cryo-EM facility at LISCB, major funder MRC.

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Citation

Lancey, C., Tehseen, M., Raducanu, V. et al. Structure of the processive human Pol δ holoenzyme. Nat Commun 11, 1109 (2020). https://doi.org/10.1038/s41467-020-14898-6

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Nature Communications

Volume

11

Issue

1

Pagination

1109

Publisher

Springer Science and Business Media LLC

issn

2041-1723

eissn

2041-1723

Acceptance date

07/02/2020

Copyright date

2020

Language

eng

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University of Leicester Publications

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