Probing the structure and function of polymerase θ helicase-like domain

Vanson, Scott; Li, Yuzhen; Wood, Richard D.; Doublié, Sylvie

doi:10.1016/j.dnarep.2022.103358

Cited by 4 publications

(2 citation statements)

References 108 publications

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“…The HLD of Pol θ is a member of the Ski2-like subfamily in helicase superfamily 2 (115). The HLD is composed of five domains, two of which are RecA-like catalytic cores (Figure 1).…”

Section: Helicase-like Domainmentioning

confidence: 99%

Genome Protection by DNA Polymerase θ

Wood

Doublié

2022

Annu. Rev. Genet.

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DNA polymerase θ (Pol θ) is a DNA repair enzyme widely conserved in animals and plants. Pol θ uses short DNA sequence homologies to initiate repair of double-strand breaks by theta-mediated end joining. The DNA polymerase domain of Pol θ is at the C terminus and is connected to an N-terminal DNA helicase–like domain by a central linker. Pol θ is crucial for maintenance of damaged genomes during development, protects DNA against extensive deletions, and limits loss of heterozygosity. The cost of using Pol θ for genome protection is that a few nucleotides are usually deleted or added at the repair site. Inactivation of Pol θ often enhances the sensitivity of cells to DNA strand–breaking chemicals and radiation. Since some homologous recombination–defective cancers depend on Pol θ for growth, inhibitors of Pol θ may be useful in treating such tumors. Expected final online publication date for the Annual Review of Genetics, Volume 56 is November 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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“…The HLD of Pol θ is a member of the Ski2-like subfamily in helicase superfamily 2 (115). The HLD is composed of five domains, two of which are RecA-like catalytic cores (Figure 1).…”

Section: Helicase-like Domainmentioning

confidence: 99%

Genome Protection by DNA Polymerase θ

Wood

Doublié

2022

Annu. Rev. Genet.

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“…POLθ-hel is involved in the ATPase dependent displacement of replication protein A (RPA) from DNA to promote TMEJ at the expense of HR. These helicase activities favor the annealing of microhomologies, thus allowing the extension of the annealed overhangs by POLθ-pol. ,− Moreover, the ability of POLθ-hel to unwind, in both ATP-dependent and -independent manner, different types of DNA substrates with 3′-5′ polarity (replication forks, blunt-ended DNA, and 3′ or 5′ overhangs) and RNA:DNA hybrids has been recently demonstrated by biochemical assays …”

Section: Introductionmentioning

confidence: 99%

Small Molecules Targeting DNA Polymerase Theta (POLθ) as Promising Synthetic Lethal Agents for Precision Cancer Therapy

et al. 2023

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Synthetic lethality (SL) is an innovative strategy in targeted anticancer therapy that exploits tumor genetic vulnerabilities. This topic has come to the forefront in recent years, as witnessed by the increased number of publications since 2007. The first proof of concept for the effectiveness of SL was provided by the approval of poly(ADP-ribose)polymerase inhibitors, which exploit a SL interaction in BRCA-deficient cells, although their use is limited by resistance. Searching for additional SL interactions involving BRCA mutations, the DNA polymerase theta (POLθ) emerged as an exciting target. This review summarizes, for the first time, the POLθ polymerase and helicase inhibitors reported to date. Compounds are described focusing on chemical structure and biological activity. With the aim to enable further drug discovery efforts in interrogating POLθ as a target, we propose a plausible pharmacophore model for POLθ-pol inhibitors and provide a structural analysis of the known POLθ ligand binding sites.

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Template-Independent Enzymatic RNA Synthesis

Karalkar,

Kent,

Tredinnick

et al. 2024

Preprint

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A route to prepare ribonucleoside triphosphates featuring a 3’-aminoxy (3’-O-NH2) removable blocking group is reported here. We then show that versions of two DNA polymerases, human DNA polymerase theta (Polθ) and mimiviral PrimPol, accept these triphosphates as substrates to add single nucleotides to an RNA primer under engineered conditions. Cleaving the O-N bond in the 3’-O-NH2group within the extended primer regenerates the 3’-OH group, facilitating subsequent polymerase cycles that add a second, selected, nucleotide. These enzymes and triphosphates together enable template-independent enzymatic RNA synthesis (TIERS) exploiting a cyclic reversible termination framework. The study shows that this process is ready for instrument adaptation by using it to add three ribonucleotides in three cycles using an engineered Polθ. This work creates a new way to synthesize RNA with a de novo defined sequence, without requiring the protecting groups, hazardous solvents, and sensitive reagents that bedevil phosphoramidite-based RNA synthesis.

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Probing the structure and function of polymerase θ helicase-like domain

Cited by 4 publications

References 108 publications

Genome Protection by DNA Polymerase θ

Genome Protection by DNA Polymerase θ

Small Molecules Targeting DNA Polymerase Theta (POLθ) as Promising Synthetic Lethal Agents for Precision Cancer Therapy

Template-Independent Enzymatic RNA Synthesis

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