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The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea

8-oxodeoxyguanosine (8-oxodG), a major oxidised base modification, has been investigated to study its impact on DNA replication in hyperthermophilic Archaea. Here we show that 8-oxodG is formed in the genome of growing cells, with elevated levels following exposure to oxidative stress. Functional ch...

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Autores principales: Killelea, Tom, Palud, Adeline, Akcha, Farida, Lemor, Mélanie, L'haridon, Stephane, Godfroy, Anne, Henneke, Ghislaine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6559790/
https://www.ncbi.nlm.nih.gov/pubmed/31184586
http://dx.doi.org/10.7554/eLife.45320
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author Killelea, Tom
Palud, Adeline
Akcha, Farida
Lemor, Mélanie
L'haridon, Stephane
Godfroy, Anne
Henneke, Ghislaine
author_facet Killelea, Tom
Palud, Adeline
Akcha, Farida
Lemor, Mélanie
L'haridon, Stephane
Godfroy, Anne
Henneke, Ghislaine
author_sort Killelea, Tom
collection PubMed
description 8-oxodeoxyguanosine (8-oxodG), a major oxidised base modification, has been investigated to study its impact on DNA replication in hyperthermophilic Archaea. Here we show that 8-oxodG is formed in the genome of growing cells, with elevated levels following exposure to oxidative stress. Functional characterisation of cell-free extracts and the DNA polymerisation enzymes, PolB, PolD, and the p41/p46 complex, alone or in the presence of accessory factors (PCNA and RPA) indicates that translesion synthesis occurs under replicative conditions. One of the major polymerisation effects was stalling, but each of the individual proteins could insert and extend past 8-oxodG with differing efficiencies. The introduction of RPA and PCNA influenced PolB and PolD in similar ways, yet provided a cumulative enhancement to the polymerisation performance of p41/p46. Overall, 8-oxodG translesion synthesis was seen to be potentially mutagenic leading to errors that are reminiscent of dA:8-oxodG base pairing.
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spelling pubmed-65597902019-06-12 The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea Killelea, Tom Palud, Adeline Akcha, Farida Lemor, Mélanie L'haridon, Stephane Godfroy, Anne Henneke, Ghislaine eLife Biochemistry and Chemical Biology 8-oxodeoxyguanosine (8-oxodG), a major oxidised base modification, has been investigated to study its impact on DNA replication in hyperthermophilic Archaea. Here we show that 8-oxodG is formed in the genome of growing cells, with elevated levels following exposure to oxidative stress. Functional characterisation of cell-free extracts and the DNA polymerisation enzymes, PolB, PolD, and the p41/p46 complex, alone or in the presence of accessory factors (PCNA and RPA) indicates that translesion synthesis occurs under replicative conditions. One of the major polymerisation effects was stalling, but each of the individual proteins could insert and extend past 8-oxodG with differing efficiencies. The introduction of RPA and PCNA influenced PolB and PolD in similar ways, yet provided a cumulative enhancement to the polymerisation performance of p41/p46. Overall, 8-oxodG translesion synthesis was seen to be potentially mutagenic leading to errors that are reminiscent of dA:8-oxodG base pairing. eLife Sciences Publications, Ltd 2019-06-11 /pmc/articles/PMC6559790/ /pubmed/31184586 http://dx.doi.org/10.7554/eLife.45320 Text en © 2019, Killelea et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry and Chemical Biology
Killelea, Tom
Palud, Adeline
Akcha, Farida
Lemor, Mélanie
L'haridon, Stephane
Godfroy, Anne
Henneke, Ghislaine
The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title_full The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title_fullStr The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title_full_unstemmed The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title_short The interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic Archaea
title_sort interplay at the replisome mitigates the impact of oxidative damage on the genetic integrity of hyperthermophilic archaea
topic Biochemistry and Chemical Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6559790/
https://www.ncbi.nlm.nih.gov/pubmed/31184586
http://dx.doi.org/10.7554/eLife.45320
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