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Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells

Apurinic/apyrimidinic (AP) sites, or abasic sites, which are a common type of endogenous DNA damage, can forge interstrand DNA–DNA cross-links via reaction with the exocyclic amino group on a nearby 2΄-deoxyguanosine or 2΄-deoxyadenosine in the opposite strand. Here, we utilized a shuttle vector met...

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Autores principales: Price, Nathan E., Li, Lin, Gates, Kent S., Wang, Yinsheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499640/
https://www.ncbi.nlm.nih.gov/pubmed/28431012
http://dx.doi.org/10.1093/nar/gkx266
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author Price, Nathan E.
Li, Lin
Gates, Kent S.
Wang, Yinsheng
author_facet Price, Nathan E.
Li, Lin
Gates, Kent S.
Wang, Yinsheng
author_sort Price, Nathan E.
collection PubMed
description Apurinic/apyrimidinic (AP) sites, or abasic sites, which are a common type of endogenous DNA damage, can forge interstrand DNA–DNA cross-links via reaction with the exocyclic amino group on a nearby 2΄-deoxyguanosine or 2΄-deoxyadenosine in the opposite strand. Here, we utilized a shuttle vector method to examine the efficiency and fidelity with which a reduced dG–AP cross-link-containing plasmid was replicated in cultured human cells. Our results showed that the cross-link constituted strong impediments to DNA replication in HEK293T cells, with the bypass efficiencies for the dG- and AP-containing strands being 40% and 20%, respectively. While depletion of polymerase (Pol) η did not perturb the bypass efficiency of the lesion, the bypass efficiency was markedly reduced (to 1–10%) in the isogenic cells deficient in Pol κ, Pol ι or Pol ζ, suggesting the mutual involvement of multiple translesion synthesis polymerases in bypassing the lesion. Additionally, replication of the cross-linked AP residue in HEK293T cells was moderately error-prone, inducing a total of ∼26% single-nucleobase substitutions at the lesion site, whereas replication past the cross-linked dG component occurred at a mutation frequency of ∼8%. Together, our results provided important insights into the effects of an AP-derived interstrand cross-link on the efficiency and accuracy of DNA replication in human cells.
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spelling pubmed-54996402017-07-10 Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells Price, Nathan E. Li, Lin Gates, Kent S. Wang, Yinsheng Nucleic Acids Res Genome Integrity, Repair and Replication Apurinic/apyrimidinic (AP) sites, or abasic sites, which are a common type of endogenous DNA damage, can forge interstrand DNA–DNA cross-links via reaction with the exocyclic amino group on a nearby 2΄-deoxyguanosine or 2΄-deoxyadenosine in the opposite strand. Here, we utilized a shuttle vector method to examine the efficiency and fidelity with which a reduced dG–AP cross-link-containing plasmid was replicated in cultured human cells. Our results showed that the cross-link constituted strong impediments to DNA replication in HEK293T cells, with the bypass efficiencies for the dG- and AP-containing strands being 40% and 20%, respectively. While depletion of polymerase (Pol) η did not perturb the bypass efficiency of the lesion, the bypass efficiency was markedly reduced (to 1–10%) in the isogenic cells deficient in Pol κ, Pol ι or Pol ζ, suggesting the mutual involvement of multiple translesion synthesis polymerases in bypassing the lesion. Additionally, replication of the cross-linked AP residue in HEK293T cells was moderately error-prone, inducing a total of ∼26% single-nucleobase substitutions at the lesion site, whereas replication past the cross-linked dG component occurred at a mutation frequency of ∼8%. Together, our results provided important insights into the effects of an AP-derived interstrand cross-link on the efficiency and accuracy of DNA replication in human cells. Oxford University Press 2017-06-20 2017-04-20 /pmc/articles/PMC5499640/ /pubmed/28431012 http://dx.doi.org/10.1093/nar/gkx266 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Genome Integrity, Repair and Replication
Price, Nathan E.
Li, Lin
Gates, Kent S.
Wang, Yinsheng
Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title_full Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title_fullStr Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title_full_unstemmed Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title_short Replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
title_sort replication and repair of a reduced 2΄-deoxyguanosine-abasic site interstrand cross-link in human cells
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5499640/
https://www.ncbi.nlm.nih.gov/pubmed/28431012
http://dx.doi.org/10.1093/nar/gkx266
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