Cargando…

A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine

During DNA replication, the presence of 8-oxoguanine (8-oxoG) lesions in the template strand cause the high-fidelity (HiFi) DNA polymerase (Pol) to stall. An early response to 8-oxoG lesions involves ‘on-the-fly’ translesion synthesis (TLS), in which a specialized TLS Pol is recruited and replaces t...

Descripción completa

Detalles Bibliográficos
Autores principales: Cranford, Matthew T, Kaszubowski, Joseph D, Trakselis, Michael A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7641752/
https://www.ncbi.nlm.nih.gov/pubmed/32997110
http://dx.doi.org/10.1093/nar/gkaa803
_version_ 1783605988251664384
author Cranford, Matthew T
Kaszubowski, Joseph D
Trakselis, Michael A
author_facet Cranford, Matthew T
Kaszubowski, Joseph D
Trakselis, Michael A
author_sort Cranford, Matthew T
collection PubMed
description During DNA replication, the presence of 8-oxoguanine (8-oxoG) lesions in the template strand cause the high-fidelity (HiFi) DNA polymerase (Pol) to stall. An early response to 8-oxoG lesions involves ‘on-the-fly’ translesion synthesis (TLS), in which a specialized TLS Pol is recruited and replaces the stalled HiFi Pol for lesion bypass. The length of TLS must be long enough for effective bypass, but it must also be regulated to minimize replication errors by the TLS Pol. The exact position where the TLS Pol ends and the HiFi Pol resumes (i.e. the length of the TLS patch) has not been described. We use steady-state and pre-steady-state kinetic assays to characterize lesion bypass intermediates formed by different archaeal polymerase holoenzyme complexes that include PCNA123 and RFC. After bypass of 8-oxoG by TLS PolY, products accumulate at the template position three base pairs beyond the lesion. PolY is catalytically poor for subsequent extension from this +3 position beyond 8-oxoG, but this inefficiency is overcome by rapid extension of HiFi PolB1. The reciprocation of Pol activities at this intermediate indicates a defined position where TLS Pol extension is limited and where the DNA substrate is handed back to the HiFi Pol after bypass of 8-oxoG.
format Online
Article
Text
id pubmed-7641752
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-76417522020-11-10 A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine Cranford, Matthew T Kaszubowski, Joseph D Trakselis, Michael A Nucleic Acids Res Genome Integrity, Repair and Replication During DNA replication, the presence of 8-oxoguanine (8-oxoG) lesions in the template strand cause the high-fidelity (HiFi) DNA polymerase (Pol) to stall. An early response to 8-oxoG lesions involves ‘on-the-fly’ translesion synthesis (TLS), in which a specialized TLS Pol is recruited and replaces the stalled HiFi Pol for lesion bypass. The length of TLS must be long enough for effective bypass, but it must also be regulated to minimize replication errors by the TLS Pol. The exact position where the TLS Pol ends and the HiFi Pol resumes (i.e. the length of the TLS patch) has not been described. We use steady-state and pre-steady-state kinetic assays to characterize lesion bypass intermediates formed by different archaeal polymerase holoenzyme complexes that include PCNA123 and RFC. After bypass of 8-oxoG by TLS PolY, products accumulate at the template position three base pairs beyond the lesion. PolY is catalytically poor for subsequent extension from this +3 position beyond 8-oxoG, but this inefficiency is overcome by rapid extension of HiFi PolB1. The reciprocation of Pol activities at this intermediate indicates a defined position where TLS Pol extension is limited and where the DNA substrate is handed back to the HiFi Pol after bypass of 8-oxoG. Oxford University Press 2020-09-30 /pmc/articles/PMC7641752/ /pubmed/32997110 http://dx.doi.org/10.1093/nar/gkaa803 Text en © The Author(s) 2020. 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
Cranford, Matthew T
Kaszubowski, Joseph D
Trakselis, Michael A
A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title_full A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title_fullStr A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title_full_unstemmed A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title_short A hand-off of DNA between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
title_sort hand-off of dna between archaeal polymerases allows high-fidelity replication to resume at a discrete intermediate three bases past 8-oxoguanine
topic Genome Integrity, Repair and Replication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7641752/
https://www.ncbi.nlm.nih.gov/pubmed/32997110
http://dx.doi.org/10.1093/nar/gkaa803
work_keys_str_mv AT cranfordmatthewt ahandoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine
AT kaszubowskijosephd ahandoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine
AT trakselismichaela ahandoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine
AT cranfordmatthewt handoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine
AT kaszubowskijosephd handoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine
AT trakselismichaela handoffofdnabetweenarchaealpolymerasesallowshighfidelityreplicationtoresumeatadiscreteintermediatethreebasespast8oxoguanine