Repriming of DNA synthesis at stalled replication forks by human PrimPol

Mourón, Silvana; Rodríguez‐Acebes, Sara; Martínez‐Jiménez, María I.; García-Gómez, Sara; Chocrón, Sandra; Blanco, Luis; Méndez, Juan Pablo

doi:10.1038/nsmb.2719

Cited by 278 publications

(443 citation statements)

References 39 publications

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“…AZT-TP is a component of the fixed-dose combinations lamivudine-zidovudine (Combivir) and abacavir sulfate-lamivudine-zidovudine (Trizivir), and CBV-TP is also a component of Trizivir, abacavir-lamivudine (Epzicom), and a recently FDA-approved combination therapy with dolutegravir and lamivudine (Triumeq). Although it may be assumed that PrimPol performs fewer incorporation events than the primary replicative nuclear polymerases ␣, ε, and ␦ and mtDNA Pol ␥, NRTI incorporation by PrimPol at lesions, including 8oxoG (15,20,21), abasic sites (15), (6-4)pp (16,21), and CPDs (16), can result in DNA breaks, genomic instability, and ultimately apoptosis (39). Our findings highlight one possible alternative mechanism of antiviral toxicity for consideration in the clinical use of these drugs and future nucleoside analog development.…”

Section: Tablementioning

confidence: 99%

“…PrimPol has emerged as a DNA and RNA primase and DNAdependent translesion synthesis (TLS) polymerase (14)(15)(16)(17)(18). A 560-amino-acid protein belonging to the archaeo-eukaryotic primase (AEP) superfamily, PrimPol possesses a conserved N-terminal AEP polymerase domain with three highly conserved catalytic motifs and a C-terminal zinc finger domain similar to the viral UL52 primase domain (17,19).…”

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confidence: 99%

“…Notably, it lacks 3=-to-5= exonuclease activity and preferentially binds the cofactor manganese (15,20). PrimPol functions in replication restart downstream of stalled replication forks (16) and can perform translesion synthesis, bypassing oxidative and UV-induced lesions, including 8-oxoguanine (8oxoG) (15,20,21), abasic sites (15), (6-4) pyrimidinepyrimidone photoproducts [(6-4)pp] (16,21), and cyclobutane pyrimidine dimers (CPDs) (16). PrimPol helps to maintain mtDNA copy numbers and replication rates and can utilize both deoxynucleoside triphosphates (dNTPs) and NTPs (15).…”

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confidence: 99%

“…Investigational in vitro studies had identified that BMS-986094 was incorporated by the human mitochondrial RNA polymerase 30-fold more efficiently than ribavirin, the then-current standard of care, with "tolerable" off-target effects (13), highlighting the importance of understanding nucleoside analog incorporation by host polymerases to prevent life-threatening adverse events. Toward this end, the NRTI incorporation efficiencies have been determined for human B-, X-, and Y-family DNA polymerases (1) while the recent discovery of PrimPol operating in the nucleus and the mitochondria suggests an unexplored mechanism of NRTI-associated toxicity (14)(15)(16)(17).…”

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Insights into the Molecular Mechanism of Polymerization and Nucleoside Reverse Transcriptase Inhibitor Incorporation by Human PrimPol

Mislak

Anderson

2016

Antimicrob Agents Chemother

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Human PrimPol is a newly identified DNA and RNA primase-polymerase of the archaeo-eukaryotic primase (AEP) superfamily and only the second known polymerase in the mitochondria. Mechanistic studies have shown that interactions of the primary mitochondrial DNA polymerase ␥ (mtDNA Pol ␥) with nucleoside reverse transcriptase inhibitors (NRTIs), key components in treating HIV infection, are a major source of NRTI-associated toxicity. Understanding the interactions of host polymerases with antiviral and anticancer nucleoside analog therapies is critical for preventing life-threatening adverse events, particularly in AIDS patients who undergo lifelong treatment. Since PrimPol has only recently been discovered, the molecular mechanism of polymerization and incorporation of natural nucleotide and NRTI substrates, crucial for assessing the potential for PrimPolmediated NRTI-associated toxicity, has not been explored. We report for the first time a transient-kinetic analysis of polymerization for each nucleotide and NRTI substrate as catalyzed by PrimPol. These studies reveal that nucleotide selectivity limits chemical catalysis while the release of the elongated DNA product is the overall rate-limiting step. Remarkably, PrimPol incorporates four of the eight FDA-approved antiviral NRTIs with a kinetic profile distinct from that of mtDNA Pol ␥ that may manifest in toxicity.N ucleoside reverse transcriptase inhibitors (NRTIs) are an important class of antivirals that target the human immunodeficiency virus (HIV) polymerase, reverse transcriptase (RT). All FDA-approved NRTIs are nucleoside analogs that lack a 3=-hydroxyl moiety to terminate DNA chain extension upon incorporation by RT into the growing proviral DNA. While significant health advances have been achieved with the use of NRTIs, the necessity for lifelong treatment to control HIV infection is limited by NRTI-associated toxicities that arise from virus-versus-host polymerase selectivity wherein NRTIs also serve as substrates for host polymerases (1, 2).The most severe NRTI-associated toxicities predominantly manifest in mitochondrial dysfunction (1-6) and are attributed primarily to incorporation by the human mitochondrial DNA polymerase ␥ (mtDNA Pol ␥) (7-9). However, there are observed discrepancies between toxicity and the potential to inhibit mtDNA Pol ␥, suggesting alternative mechanisms and evaluation of additional host cell polymerases as potential perpetrators of antiviral toxicity (10, 11). Understanding the propensity for host cell polymerases to incorporate nucleoside analogs is critical for assessing safety in the design and development of antiviral, -parasitic, -bacterial, and -cancer nucleoside analog therapies. For instance, development of the antiviral ribonucleoside triphosphate analog BMS-986094 for the treatment of hepatitis C virus was halted in phase II after nine patients were hospitalized and one died (12). Investigational in vitro studies had identified that BMS-986094 was incorporated by the human mitochondrial RNA polymerase 30-fold more ...

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Section: Tablementioning

confidence: 99%

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Insights into the Molecular Mechanism of Polymerization and Nucleoside Reverse Transcriptase Inhibitor Incorporation by Human PrimPol

Mislak

Anderson

2016

Antimicrob Agents Chemother

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“…28 A recent study has identified a new TLS polymerase, PrimPol, which function in lesion bypass during replication is dNTP dependent. 29 In the future, it would be interesting to understand how the DNA polymerase is selected to adapt low dNTP pools for the gap-filling step in the NER process in quiescent cells.…”

Section: Discussionmentioning

confidence: 99%

The contribution of CMP kinase to the efficiency of DNA repair

et al. 2015

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Abbreviations: dNTP, deoxynucleoside triphosphates; CMPK, CMP/UMP kinase; RNR, ribonucleotide reductase; 6-4 pp, photoproduct; R1C, C-terminal fragment of R1 subunit of RNR.Cellular supply of deoxynucleoside triphosphates (dNTPs) is crucial for DNA replication and repair. In this study, we investigated the role of CMP/UMP kinase (CMPK), an enzyme catalyzes CDP formation, in DNA repair. Knockdown of CMPK delays DNA repair during recovery from UV damage in serum-deprived cells but not in the cells without serum deprivation. Exogenous supply of cytidine or deoxycytidine facilitates DNA repair dependent on CMPK in serumdeprived cells, suggesting that the synthesis of dCDP or CDP determines the rate of repair. However, CMPK knockdown does not affect the steady state level of dCTP in serum-deprived cells. We then found the localization of CMPK at DNA damage sites and its complex formation with Tip60 and ribonucleotide reductase. Our analysis demonstrated that the N-terminal 32-amino-acid of CMPK is required for its recruitment to DNA damage sites in a Tip60-dependent manner. Re-expression of wild-type but not N-terminus deleted CMPK restores the efficiency of DNA repair in CMPK knockdown cells. We proposed that site-specific dCDP formation via CMPK provides a means to facilitate DNA repair in serumdeprived cells.

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Manipulating Intratumoral Fenton Chemistry for Enhanced Chemodynamic and Chemodynamic‐Synergized Multimodal Therapy

et al. 2021

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Chemodynamic therapy (CDT) uses the tumor microenvironment‐assisted intratumoral Fenton reaction for generating highly toxic hydroxyl free radicals (•OH) to achieve selective tumor treatment. However, the limited intratumoral Fenton reaction efficiency restricts the therapeutic efficacy of CDT. Recent years have witnessed the impressive development of various strategies to increase the efficiency of intratumoral Fenton reaction. The introduction of these reinforcement strategies can dramatically improve the treatment efficiency of CDT and further promote the development of enhanced CDT (ECDT)‐based multimodal anticancer treatments. In this review, the authors systematically introduce these reinforcement strategies, from their basic working principles, reinforcement mechanisms to their representative clinical applications. Then, ECDT‐based multimodal anticancer therapy is discussed, including how to integrate these emerging Fenton reinforcement strategies for accelerating the development of multimodal anticancer therapy, as well as the synergistic mechanisms of ECDT and other treatment methods. Eventually, future direction and challenges of ECDT and ECDT‐based multimodal synergistic therapies are elaborated, highlighting the key scientific problems and unsolved technical bottlenecks to facilitate clinical translation.

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Repriming of DNA synthesis at stalled replication forks by human PrimPol

Cited by 278 publications

References 39 publications

Insights into the Molecular Mechanism of Polymerization and Nucleoside Reverse Transcriptase Inhibitor Incorporation by Human PrimPol

Insights into the Molecular Mechanism of Polymerization and Nucleoside Reverse Transcriptase Inhibitor Incorporation by Human PrimPol

The contribution of CMP kinase to the efficiency of DNA repair

Manipulating Intratumoral Fenton Chemistry for Enhanced Chemodynamic and Chemodynamic‐Synergized Multimodal Therapy

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