Human DNA polymerase   possesses 5'-dRP lyase activity and functions in single-nucleotide base excision repair in vitro

Prasad, Rajendra; Longley, Matthew J.; Sharief, Farida S.; Hou, Esther W.; Copeland, William C.; Wilson, Samuel H.

doi:10.1093/nar/gkp035

Cited by 94 publications

(104 citation statements)

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“…Finally, the persistence of DPCs after DNA strand scission is also consistent with relatively slow Schiff base hydrolysis. Although a number of base excision repair proteins and DNA polymerases exhibit lyase activities toward AP sites (39)(40)(41)(42), it is less common for proteins that are not enzymes to react with DNA. In a recent rare example of the latter, Ku excised AP sites from DNA via a Schiff base intermediate (43).…”

Section: Discussionmentioning

confidence: 99%

Rapid DNA–protein cross-linking and strand scission by an abasic site in a nucleosome core particle

Sczepanski

Wong

McKnight

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

175

316

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Apurinic/apyrimidinic (AP) sites are ubiquitous DNA lesions that are highly mutagenic and cytotoxic if not repaired. In addition, clusters of two or more abasic lesions within one to two turns of DNA, a hallmark of ionizing radiation, are repaired much less efficiently and thus present greater mutagenic potential. Abasic sites are chemically labile, but naked DNA containing them undergoes strand scission slowly with a half-life on the order of weeks. We find that independently generated AP sites within nucleosome core particles are highly destabilized, with strand scission occurring ∼60-fold more rapidly than in naked DNA. The majority of core particles containing single AP lesions accumulate DNA-protein cross-links, which persist following strand scission. The N-terminal region of histone protein H4 contributes significantly to DNA-protein crosslinks and strand scission when AP sites are produced approximately 1.5 helical turns from the nucleosome dyad, which is a known hot spot for nucleosomal DNA damage. Reaction rates for AP sites at two positions within this region differ by ∼4-fold. However, the strand scission of the slowest reacting AP site is accelerated when it is part of a repair resistant bistranded lesion composed of two AP sites, resulting in rapid formation of double strand breaks in high yields. Multiple lysine residues within a single H4 protein catalyze double strand cleavage through a mechanism believed to involve a templating effect. These results show that AP sites within the nucleosome produce significant amounts of DNA-protein cross-links and generate double strand breaks, the most deleterious form of DNA damage.chromatin | oxidative damage | histone modification E xogenously and endogenously produced agents continuously damage DNA. More than 70 types of damage have been identified, and significant effort is expended to determine which of these lesions are biologically important (1, 2). The apurinic/apyrimidinic (AP) site is a ubiquitous form of DNA damage that is produced in excess of 10,000 lesions per cell each day (3). This highly mutagenic lesion results from spontaneous hydrolysis of native and damaged nucleotides. AP sites are also formed as intermediates during base excision repair of alkylated and oxidized nucleotides and are themselves removed by multiple enzymatic pathways (4, 5). In mammalian cells incision of the AP's 5′-phosphate by Ape1, followed by excision of the resulting 5′-terminal 2′-deoxyribose phosphate by DNA polymerase β is the major pathway for removing the lesion. Redundant repair pathways are reflective of the high mutagenic potential and large quantities of AP produced and are indicative of their physiological significance. Mammalian cells lacking Ape1, the major base excision repair protein responsible for incising AP sites, are embryonic lethal (6). Recent observations reinforce the perception that AP sites are cytotoxic. For instance, formation of bursts of AP sites by the natural product leinamycin is postulated to be the source of this antitumor agent's cy...

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Section: Discussionmentioning

confidence: 99%

Rapid DNA–protein cross-linking and strand scission by an abasic site in a nucleosome core particle

Sczepanski

Wong

McKnight

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

175

316

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“…Nevertheless, we cannot rule out the possibility that mechanisms other than DNA replication could be involved. Excess POLQ could titrate cofactors that are involved in TLS or DNA repair pathways, such as alternative nonhomologous end joining, ICL repair, or BER, where this polymerase might be involved (19)(20)(21)(22)(23). Shorter DNA replication tracts and a modest sensitivity to alkylating agents in POLQ-overexpressing cells could therefore result from an accumulation of DNA damage due to these defective DNA transactions.…”

Section: Discussionmentioning

confidence: 99%

“…Because POLQ has been proposed to play a role in BER and translesion synthesis (TLS) (19,21,22,25), we hypothesized that overexpression of POLQ could interfere with the cellular tolerance to DNA damage (including damage generated by reactive oxygen species), thus leading to reduced kinetics of replication fork progression. Therefore, we measured the sensitivity of the POLQ overexpressing MRC5-SV cells to increasing doses of methylmethane-sulfonate (MMS) and N-Nitroso-N-Methylurea (MNU) (Fig.…”

Section: Polq Overexpressing Cells Display Spontaneous Chromosomementioning

confidence: 99%

“…The observation that chaos1 mutant mice display enhanced micronuclei in red blood cells further suggests a role of POLQ in maintaining genomic stability in the absence of any exogenous stress. In addition it seems to be involved in tolerance of bulky adducts or in DNA repair pathways such as the base-excision repair (BER), DNA interstrand cross-link (ICL), and DNA break repair pathways (19)(20)(21)(22)(23). To investigate the molecular consequences of POLQ up-regulation, we generated human MRC5-SV cells that stably overexpressed the POLQ protein.…”

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confidence: 99%

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DNA polymeraseθ up-regulation is associated with poor survival in breast cancer, perturbs DNA replication, and promotes genetic instability

Lemée

Bergoglio

Fernandez-Vidal

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

179

174

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"Replicative stress" is one of the main factors underlying neoplasia from its early stages. Genes involved in DNA synthesis may therefore represent an underexplored source of potential prognostic markers for cancer. To this aim, we generated gene expression profiles from two independent cohorts (France, n = 206; United Kingdom, n = 117) of patients with previously untreated primary breast cancers. We report here that among the 13 human nuclear DNA polymerase genes, DNA Polymerase θ (POLQ) is the only one significantly up-regulated in breast cancer compared with normal breast tissues. Importantly, POLQ up-regulation significantly correlates with poor clinical outcome (4.3-fold increased risk of death in patients with high POLQ expression), and this correlation is independent of Cyclin E expression or the number of positive nodes, which are currently considered as markers for poor outcome. POLQ expression provides thus an additional indicator for the survival outcome of patients with high Cyclin E tumor expression or high number of positive lymph nodes. Furthermore, to decipher the molecular consequences of POLQ up-regulation in breast cancer, we generated human MRC5-SV cell lines that stably overexpress POLQ. Strong POLQ expression was directly associated with defective DNA replication fork progression and chromosomal damage. Therefore, POLQ overexpression may be a promising genetic instability and prognostic marker for breast cancer.specialized DNA replication | prognosis marker | S-phase checkpoint B esides the "replicative" DNA polymerases POLA, POLD, and POLE, which are involved in conventional DNA replication of the undamaged genome, mammalian nuclei contain 10 additional specialized DNA polymerases that play a role in replication, repair, and recombination of damaged DNA (1, 2) and thus may be of paramount importance to preserve the integrity of the genome.Specialized DNA polymerases are frequently deregulated in neoplasia (3-10). Indeed, the intracellular balance between the error-free, replicative polymerases POLA, POLD, and POLE and the error-prone, specialized DNA polymerases (POLH, POLL, POLM, POLN, POLK, POLB, POLI, POLQ, POLZ/REV3L, and REV1) appears to be of great importance for the maintenance of genome stability (11)(12)(13)(14). Here, we wondered whether misregulation of DNA polymerases could be a signature of breast cancer progression. Indeed, beside the standard classification used by pathologists, there is a clear lack of tools to accurately predict the clinical outcome of many patients.We specifically measured the expression levels of the 13 human replicative and specialized DNA polymerases in 206 breast carcinomas. We report that, differently from the replicative and the other specialized DNA polymerases, POLQ was significantly upregulated in most of the breast tumors analyzed. Such up-regulation was associated with poor clinical outcome.POLQ is an error-prone, specialized DNA polymerase that might operate during "normal" genomic replication because it bypasses some endogenous DNA lesions an...

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“…Importantly, Pol n both physically and functionally interacts with the FA pathway (Moldovan et al 2010). Additionally, Drosophila Mus308 is involved in microhomology-mediated NHEJ (Chan et al 2010;Yu and McVey 2010) and Pol u in BER (Yoshimura et al 2006;Prasad et al 2009), which is likely facilitated by its terminal-transferase (Hogg et al 2012) and dRP-lyase (Prasad et al 2009) activities. As with the X-family polymerases, it seems likely that these roles are important in contexts involving complex or clustered damage.…”

Section: Translesion Synthesis and Mutagenesis By Eukaryotic X-familymentioning

confidence: 99%

Translesion DNA Synthesis and Mutagenesis in Eukaryotes

Sale

2013

Cold Spring Harbor Perspectives in Biology

224

231

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The structural features that enable replicative DNA polymerases to synthesize DNA rapidly and accurately also limit their ability to copy damaged DNA. Direct replication of DNA damage is termed translesion synthesis (TLS), a mechanism conserved from bacteria to mammals and executed by an array of specialized DNA polymerases. This chapter examines how these translesion polymerases replicate damaged DNA and how they are regulated to balance their ability to replicate DNA lesions with the risk of undesirable mutagenesis. It also discusses how TLS is co-opted to increase the diversity of the immunoglobulin gene hypermutation and the contribution it makes to the mutations that sculpt the genome of cancer cells.

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Human DNA polymerase possesses 5'-dRP lyase activity and functions in single-nucleotide base excision repair in vitro

Cited by 94 publications

References 34 publications

Rapid DNA–protein cross-linking and strand scission by an abasic site in a nucleosome core particle

Rapid DNA–protein cross-linking and strand scission by an abasic site in a nucleosome core particle

DNA polymeraseθ up-regulation is associated with poor survival in breast cancer, perturbs DNA replication, and promotes genetic instability

Translesion DNA Synthesis and Mutagenesis in Eukaryotes

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