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PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells

Eukaryotic Primase-Polymerase (PrimPol) is an enzyme that maintains efficient DNA duplication by repriming replication restart downstream of replicase stalling lesions and structures. To elucidate the cellular requirements for PrimPol in human cells, we generated PrimPol-deleted cell lines and show...

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Autores principales: Bailey, Laura J, Bianchi, Julie, Doherty, Aidan J
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486543/
https://www.ncbi.nlm.nih.gov/pubmed/30715459
http://dx.doi.org/10.1093/nar/gkz056
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author Bailey, Laura J
Bianchi, Julie
Doherty, Aidan J
author_facet Bailey, Laura J
Bianchi, Julie
Doherty, Aidan J
author_sort Bailey, Laura J
collection PubMed
description Eukaryotic Primase-Polymerase (PrimPol) is an enzyme that maintains efficient DNA duplication by repriming replication restart downstream of replicase stalling lesions and structures. To elucidate the cellular requirements for PrimPol in human cells, we generated PrimPol-deleted cell lines and show that it plays key roles in maintaining active replication in both the nucleus and mitochondrion, even in the absence of exogenous damage. Human cells lacking PrimPol exhibit delayed recovery after UV-C damage and increased mutation frequency, micronuclei and sister chromatin exchanges but are not sensitive to genotoxins. PrimPol is also required during mitochondrial replication, with PrimPol-deficient cells having increased mtDNA copy number but displaying a significant decrease in replication. Deletion of PrimPol in XPV cells, lacking functional polymerase Eta, causes an increase in DNA damage sensitivity and pronounced fork stalling after UV-C treatment. We show that, unlike canonical TLS polymerases, PrimPol is important for allowing active replication to proceed, even in the absence of exogenous damage, thus preventing the accumulation of excessive fork stalling and genetic mutations. Together, these findings highlight the importance of PrimPol for maintaining efficient DNA replication in unperturbed cells and its complementary roles, with Pol Eta, in damage tolerance in human cells.
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spelling pubmed-64865432019-05-01 PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells Bailey, Laura J Bianchi, Julie Doherty, Aidan J Nucleic Acids Res Genome Integrity, Repair and Replication Eukaryotic Primase-Polymerase (PrimPol) is an enzyme that maintains efficient DNA duplication by repriming replication restart downstream of replicase stalling lesions and structures. To elucidate the cellular requirements for PrimPol in human cells, we generated PrimPol-deleted cell lines and show that it plays key roles in maintaining active replication in both the nucleus and mitochondrion, even in the absence of exogenous damage. Human cells lacking PrimPol exhibit delayed recovery after UV-C damage and increased mutation frequency, micronuclei and sister chromatin exchanges but are not sensitive to genotoxins. PrimPol is also required during mitochondrial replication, with PrimPol-deficient cells having increased mtDNA copy number but displaying a significant decrease in replication. Deletion of PrimPol in XPV cells, lacking functional polymerase Eta, causes an increase in DNA damage sensitivity and pronounced fork stalling after UV-C treatment. We show that, unlike canonical TLS polymerases, PrimPol is important for allowing active replication to proceed, even in the absence of exogenous damage, thus preventing the accumulation of excessive fork stalling and genetic mutations. Together, these findings highlight the importance of PrimPol for maintaining efficient DNA replication in unperturbed cells and its complementary roles, with Pol Eta, in damage tolerance in human cells. Oxford University Press 2019-05-07 2019-01-31 /pmc/articles/PMC6486543/ /pubmed/30715459 http://dx.doi.org/10.1093/nar/gkz056 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Genome Integrity, Repair and Replication
Bailey, Laura J
Bianchi, Julie
Doherty, Aidan J
PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title_full PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title_fullStr PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title_full_unstemmed PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title_short PrimPol is required for the maintenance of efficient nuclear and mitochondrial DNA replication in human cells
title_sort primpol is required for the maintenance of efficient nuclear and mitochondrial dna replication in human cells
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6486543/
https://www.ncbi.nlm.nih.gov/pubmed/30715459
http://dx.doi.org/10.1093/nar/gkz056
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