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Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA
Replicative DNA polymerases (DNAPs) have evolved the ability to copy the genome with high processivity and fidelity. In Eukarya and Archaea, the processivity of replicative DNAPs is greatly enhanced by its binding to the proliferative cell nuclear antigen (PCNA) that encircles the DNA. We determined...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7101311/ https://www.ncbi.nlm.nih.gov/pubmed/32221299 http://dx.doi.org/10.1038/s41467-020-15392-9 |
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author | Madru, Clément Henneke, Ghislaine Raia, Pierre Hugonneau-Beaufet, Inès Pehau-Arnaudet, Gérard England, Patrick Lindahl, Erik Delarue, Marc Carroni, Marta Sauguet, Ludovic |
author_facet | Madru, Clément Henneke, Ghislaine Raia, Pierre Hugonneau-Beaufet, Inès Pehau-Arnaudet, Gérard England, Patrick Lindahl, Erik Delarue, Marc Carroni, Marta Sauguet, Ludovic |
author_sort | Madru, Clément |
collection | PubMed |
description | Replicative DNA polymerases (DNAPs) have evolved the ability to copy the genome with high processivity and fidelity. In Eukarya and Archaea, the processivity of replicative DNAPs is greatly enhanced by its binding to the proliferative cell nuclear antigen (PCNA) that encircles the DNA. We determined the cryo-EM structure of the DNA-bound PolD–PCNA complex from Pyrococcus abyssi at 3.77 Å. Using an integrative structural biology approach — combining cryo-EM, X-ray crystallography, protein–protein interaction measurements, and activity assays — we describe the molecular basis for the interaction and cooperativity between a replicative DNAP and PCNA. PolD recruits PCNA via a complex mechanism, which requires two different PIP-boxes. We infer that the second PIP-box, which is shared with the eukaryotic Polα replicative DNAP, plays a dual role in binding either PCNA or primase, and could be a master switch between an initiation and a processive phase during replication. |
format | Online Article Text |
id | pubmed-7101311 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-71013112020-03-30 Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA Madru, Clément Henneke, Ghislaine Raia, Pierre Hugonneau-Beaufet, Inès Pehau-Arnaudet, Gérard England, Patrick Lindahl, Erik Delarue, Marc Carroni, Marta Sauguet, Ludovic Nat Commun Article Replicative DNA polymerases (DNAPs) have evolved the ability to copy the genome with high processivity and fidelity. In Eukarya and Archaea, the processivity of replicative DNAPs is greatly enhanced by its binding to the proliferative cell nuclear antigen (PCNA) that encircles the DNA. We determined the cryo-EM structure of the DNA-bound PolD–PCNA complex from Pyrococcus abyssi at 3.77 Å. Using an integrative structural biology approach — combining cryo-EM, X-ray crystallography, protein–protein interaction measurements, and activity assays — we describe the molecular basis for the interaction and cooperativity between a replicative DNAP and PCNA. PolD recruits PCNA via a complex mechanism, which requires two different PIP-boxes. We infer that the second PIP-box, which is shared with the eukaryotic Polα replicative DNAP, plays a dual role in binding either PCNA or primase, and could be a master switch between an initiation and a processive phase during replication. Nature Publishing Group UK 2020-03-27 /pmc/articles/PMC7101311/ /pubmed/32221299 http://dx.doi.org/10.1038/s41467-020-15392-9 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Madru, Clément Henneke, Ghislaine Raia, Pierre Hugonneau-Beaufet, Inès Pehau-Arnaudet, Gérard England, Patrick Lindahl, Erik Delarue, Marc Carroni, Marta Sauguet, Ludovic Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title | Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title_full | Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title_fullStr | Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title_full_unstemmed | Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title_short | Structural basis for the increased processivity of D-family DNA polymerases in complex with PCNA |
title_sort | structural basis for the increased processivity of d-family dna polymerases in complex with pcna |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7101311/ https://www.ncbi.nlm.nih.gov/pubmed/32221299 http://dx.doi.org/10.1038/s41467-020-15392-9 |
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