In vitro lesion bypass by human PrimPol

Makarova, Аlena V.; Boldinova, Elizaveta O.; Belousova, E. A.; Lavrik, Olga I.

doi:10.1016/j.dnarep.2018.07.009

Cited by 30 publications

(27 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The DNA damage tolerance pathways, where PrimPol is involved, permit lesion bypass during DNA synthesis that can be carried through translesion synthesis ( 132 , 133 ). Thereby, the sliding clamp PCNA serves as a polymerase processivity factor.…”

Section: Rpa As a Replisome Componentmentioning

confidence: 99%

Replication protein A: a multifunctional protein with roles in DNA replication, repair and beyond

Dueva

Iliakis

2020

NAR Cancer

View full text Add to dashboard Cite

Single-stranded DNA (ssDNA) forms continuously during DNA replication and is an important intermediate during recombination-mediated repair of damaged DNA. Replication protein A (RPA) is the major eukaryotic ssDNA-binding protein. As such, RPA protects the transiently formed ssDNA from nucleolytic degradation and serves as a physical platform for the recruitment of DNA damage response factors. Prominent and well-studied RPA-interacting partners are the tumor suppressor protein p53, the RAD51 recombinase and the ATR-interacting proteins ATRIP and ETAA1. RPA interactions are also documented with the helicases BLM, WRN and SMARCAL1/HARP, as well as the nucleotide excision repair proteins XPA, XPG and XPF–ERCC1. Besides its well-studied roles in DNA replication (restart) and repair, accumulating evidence shows that RPA is engaged in DNA activities in a broader biological context, including nucleosome assembly on nascent chromatin, regulation of gene expression, telomere maintenance and numerous other aspects of nucleic acid metabolism. In addition, novel RPA inhibitors show promising effects in cancer treatment, as single agents or in combination with chemotherapeutics. Since the biochemical properties of RPA and its roles in DNA repair have been extensively reviewed, here we focus on recent discoveries describing several non-canonical functions.

show abstract

Section: Rpa As a Replisome Componentmentioning

confidence: 99%

Replication protein A: a multifunctional protein with roles in DNA replication, repair and beyond

Dueva

Iliakis

2020

NAR Cancer

View full text Add to dashboard Cite

show abstract

“…Thymine glycol (Tg) is a strong block or replicative DNAPs 42 . Two independent groups have shown that HsPrimPol is blocked at a Tg using Mg ++ as cofactor 22 , 43 . However, HsPrimPol bypasses a Tg lesion using Mn ++ (41).…”

Section: Resultsmentioning

confidence: 99%

Arabidopsis thaliana PrimPol is a primase and lesion bypass DNA polymerase with the biochemical characteristics to cope with DNA damage in the nucleus, mitochondria, and chloroplast

García‐Medel

Peralta-Castro

Baruch‐Torres

et al. 2021

Sci Rep

View full text Add to dashboard Cite

PrimPol is a novel Primase–Polymerase that synthesizes RNA and DNA primers de novo and extents from these primers as a DNA polymerase. Animal PrimPol is involved in nuclear and mitochondrial DNA replication by virtue of its translesion DNA synthesis (TLS) and repriming activities. Here we report that the plant model Arabidopsis thaliana encodes a functional PrimPol (AtPrimPol). AtPrimPol is a low fidelity and a TLS polymerase capable to bypass DNA lesions, like thymine glycol and abasic sites, by incorporating directly across these lesions or by skipping them. AtPrimPol is also an efficient primase that preferentially recognizes the single-stranded 3′-GTCG-5′ DNA sequence, where the 3′-G is cryptic. AtPrimPol is the first DNA polymerase that localizes in three cellular compartments: nucleus, mitochondria, and chloroplast. In vitro, AtPrimPol synthesizes primers that are extended by the plant organellar DNA polymerases and this reaction is regulated by organellar single-stranded binding proteins. Given the constant exposure of plants to endogenous and exogenous DNA-damaging agents and the enzymatic capabilities of lesion bypass and re-priming of AtPrimPol, we postulate a predominant role of this enzyme in avoiding replication fork collapse in all three plant genomes, both as a primase and as a TLS polymerase.

show abstract

“…Early studies showed that thymine glycols, which comprise up to 30% of the H 2 O 2 -induced base damage, represent strong blocks to E. coli DNA polymerase I and phage T4 DNA polymerase in vitro ( Ide et al 1985 ; Rouet and Essigmann 1985 ; Clark and Beardsley 1986 ; Hayes and Leclerc 1986 ; Blakely et al 1990 ), and it was often inferred that this would extend generally to other DNA polymerases. However, since these initial studies, several other polymerases have been examined and found to efficiently bypass this lesion, including many in humans ( Fischhaber et al 2002 ; Takata et al 2006 ; Belousova et al 2010 ; Yoon et al 2010 ; Hogg et al 2011 ; Makarova et al 2018 ). Further, auxiliary proteins associated with the replisome, such as single-strand binding protein and processivity factors, can increase bypass efficiency in some cases ( Maga et al 2007 ; McCulloch et al 2009 ).…”

Section: Discussionmentioning

confidence: 99%

Events associated with DNA replication disruption are not observed in hydrogen peroxide-treated Escherichia coli

Hoff

Schmidt

Hackert

et al. 2021

G3 Genes|Genomes|Genetics

View full text Add to dashboard Cite

UV irradiation induces pyrimidine dimers that block polymerases and disrupt the replisome. Restoring replication depends on the recF pathway proteins which process and maintain the replication fork DNA to allow the lesion to be repaired before replication resumes. Oxidative DNA lesions, such as those induced by hydrogen peroxide (H2O2), are often thought to require similar processing events, yet far less is known about how cells process oxidative damage during replication. Here we show that replication is not disrupted by H2O2-induced DNA damage in vivo. Following an initial inhibition, replication resumes in the absence of either lesion removal or RecF-processing. Restoring DNA synthesis depends on the presence of manganese in the medium, which we show is required for replication, but not repair to occur. The results demonstrate that replication is enzymatically inactivated, rather than physically disrupted by H2O2-induced DNA damage; indicate that inactivation is likely caused by oxidation of an iron-dependent replication or replication-associated protein that requires manganese to restore activity and synthesis; and address a long standing paradox as to why oxidative glycosylase mutants are defective in repair, yet not hypersensitive to H2O2. The oxygen-sensitive pausing may represent an adaptation that prevents replication from occurring under potentially lethal or mutagenic conditions.

show abstract

In vitro lesion bypass by human PrimPol

Cited by 30 publications

References 34 publications

Replication protein A: a multifunctional protein with roles in DNA replication, repair and beyond

Replication protein A: a multifunctional protein with roles in DNA replication, repair and beyond

Arabidopsis thaliana PrimPol is a primase and lesion bypass DNA polymerase with the biochemical characteristics to cope with DNA damage in the nucleus, mitochondria, and chloroplast

Events associated with DNA replication disruption are not observed in hydrogen peroxide-treated Escherichia coli

Contact Info

Product

Resources

About