4.7 Review

Multifaceted Nature of DNA Polymerase θ

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Structural basis of DNA polymerase θ mediated DNA end joining

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Summary: DNA polymerase theta (Pol theta) plays a crucial role in the microhomology-mediated end joining (MMEJ) pathway for repairing DNA double-strand breaks. This study presents cryo-electron microscope structures of Lates calcarifer Pol theta, revealing its interactions with long and short DNA substrates and its similarity to high-fidelity A-family polymerases. Computational simulations and mutagenesis studies suggest that unique insertion loops of Pol theta stabilize short DNA binding and assemble the active site for MMEJ repair. These findings provide a structural basis for understanding Pol theta-mediated MMEJ.

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Summary: DNA polymerase theta (Pol theta)-mediated end joining is a recently characterized DNA repair pathway that functions in various cellular contexts to repair DNA double-strand breaks that are not repaired by other pathways. Pol theta-mediated end joining both helps maintain the genome and causes genome instability, and is an emerging therapeutic target in cancer.

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Templated Insertions Are Associated Specifically with BRCA2 Deficiency and Overall Survival in Advanced Ovarian Cancer

Grace Moore et al.

Summary: This study identified small, templated insertions as genomic signatures indicative of theta-mediated end-joining in tumors with pathogenic biallelic alterations in core HR genes. These signatures can serve as prognostic tools to assess patient response to therapies targeting HR deficiency.

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Summary: The DNA damage response is a complex protein network that detects and repairs DNA damage, preventing cancer. PARP inhibitors are effective in treating cancer, and new DDR targets are being investigated. Drug resistance is a problem in cancer treatment, and there is some understanding of PARPi resistance.

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Polymerase theta-helicase promotes end joining by stripping single-stranded DNA-binding proteins and bridging DNA ends

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Summary: Pol theta-h is a highly processive single-stranded DNA motor protein that can strip Replication Protein A from ssDNA and disassemble RAD51 filaments. It can also bridge two non-complementary DNA strands, aiding in polymerization by the Pol theta polymerase domain.

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POLO prevents MRE11-NBS1-CtIP-dependent fork breakage in the absence of BRCA2/RAD51 by filling lagging-strand gaps

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Next-Generation Sequencing-Based Analysis of the Roles of DNA Polymerases ν and θ in the Replicative Bypass of 8-Oxo-7,8-dihydroguanine in Human Cells

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Summary: Pols nu and theta play important roles in the replicative bypass of 8-oxoG in human cells, with Pol nu playing a key role in translesion synthesis of 8-oxoG.

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Poly(ADP) ribose polymerase promotes DNA polymerase theta-mediated end joining by activation of end resection

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Summary: The DNA polymerase theta (Pol theta)-mediated end joining (TMEJ) pathway is essential for repair of chromosomal double strand breaks (DSBs). PARP activity promotes TMEJ by stimulating resection, but its effect is limited.

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Genome Protection by DNA Polymerase θ

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Summary: DNA polymerase theta (Pol theta) is a DNA repair enzyme that plays a crucial role in maintaining genome integrity and protecting DNA against deletions and loss of heterozygosity. However, the use of Pol theta for genome protection may result in the deletion or addition of a few nucleotides at the repair site. Inactivation of Pol theta can enhance cell sensitivity to DNA strand-breaking chemicals and radiation, making inhibitors of Pol theta potentially useful in treating cancers dependent on homologous recombination.

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Summary: Pancreatic ductal adenocarcinoma (PDAC) is a deadly cancer, and the oncogenic KRAS mutation plays a key role in its development. Our study shows that KRAS mutation increases the activity of the error-prone alt-EJ repair mechanism, and depletion of Pol theta delays the development of pancreatic cancer and improves survival.

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POLQ suppresses genome instability and alterations in DNA repeat tract lengths

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Summary: DNA polymerase theta (POLQ) is a key enzyme involved in the repair of DNA double-strand breaks. Research has shown that cells lacking POLQ are resistant to growth deceleration during telomere-driven crisis, and exhibit longer telomeres and increased genetic heterogeneity.

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Discovery, Characterization, and Structure-Based Optimization of Small-Molecule In Vitro and In Vivo Probes for Human DNA Polymerase Theta

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Summary: This article describes the discovery and characterization of selective small molecule probes for human DNA polymerase theta (Pol0), a key enzyme involved in microhomology-mediated DNA repair. The crystallographic data provides insight into the unique mechanism of inhibition of these compounds, which could be valuable for the treatment of BRCA deficient and other DNA repair pathway defective cancers.

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Identification of RP-6685, an Orally Bioavailable Compound that Inhibits the DNA Polymerase Activity of Polθ

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Summary: This study describes the search for a selective inhibitor of DNA polymerase theta and the successful identification of RP-6685 as a potent, selective, and orally bioavailable inhibitor with potential efficacy in breast and ovarian cancer treatment.

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EMT Transcription Factor ZEB1 Represses the Mutagenic POLθ-Mediated End-Joining Pathway in Breast Cancers

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Summary: The study demonstrated that ZEB1 represses the expression of POLQ, inhibiting TMEJ activity and controlling genome integrity in breast cancer cells. This reveals an original mechanism of TMEJ regulation and highlights ZEB1 as a key player in maintaining genome stability during cancer progression.

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Human DNA polymerase θ harbors DNA end-trimming activity critical for DNA repair

Karl E. Zahn et al.

Summary: The study reveals that DNA polymerase theta (pol theta) possesses a nuclease activity, distinct from its DNA synthesis activity, which plays a crucial role in the repair of DNA double-strand breaks. This nuclease activity requires metal ions, specific DNA nucleotides, and can trim or extend DNA based on the DNA repair context.

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Polθ promotes the repair of 5′-DNA-protein crosslinks by microhomology-mediated end-joining

Gurushankar Chandramouly et al.

Summary: By studying Pol theta repair of DSBs carrying DPCs, it was found that Pol theta facilitates the repair of these DNA lesions through MMEJ, particularly in repairing 5' terminal DPCs. The studies also revealed that dual deficiency in Pol theta and TDP2 causes increased cellular sensitivity to etoposide, highlighting MMEJ as an alternative pathway for DPC repair.

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Recurrent deletions in clonal hematopoiesis are driven by microhomology-mediated end joining

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DNA polymerase theta suppresses mitotic crossing over

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Summary: Chromosome breaks pose a common threat to DNA stability, and mutations in BRCA1 and BRCA2 genes can lead to hereditary breast cancer. Polymerase theta plays a crucial role in homologous recombination pathway, particularly in relation to SLX4 and GEN1 proteins.

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Knockdown of POLQ interferes the development and progression of hepatocellular carcinoma through regulating cell proliferation, apoptosis and migration

Qi Pan et al.

Summary: The study revealed that knockdown of POLQ in hepatocellular carcinoma cells led to decreased proliferation and migration, increased apoptosis, and a higher percentage of cells in G2 phase. Xenografts in mice implanted with shPOLQ cells grew slower and expressed lower levels of Ki67 compared to shCtrl cells. Downstream signaling pathways related to apoptosis showed enhanced phosphorylation of HSP27 and JNK, while mTOR, PRAS40, ERK1/2, and STAT3 pathways were deactivated.

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Polθ inhibitors elicit BRCA-gene synthetic lethality and target PARP inhibitor resistance

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Summary: The study identifies small molecule inhibitors targeting Pol theta polymerase activity that induce BRCA1/2 synthetic lethality, enhance PARP inhibitor effects, and address PARP inhibitor resistance caused by defects in the 53BP1/Shieldin pathway.

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DNA Polymerase θ: A Cancer Drug Target with Reverse Transcriptase Activity

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Summary: The discovery of Pol theta as a synthetic lethal enzyme with HR factors and other DNA repair proteins has generated significant interest in its potential as an anti-cancer drug target. Its activities in microhomology-mediated end-joining (MMEJ) and translesion synthesis (TLS) have opened new possibilities for therapeutic interventions against genotoxic cancer therapies. Additionally, recent findings revealing Pol theta as a naturally occurring reverse transcriptase (RT) in mammalian cells have provided further insights into its diverse functions.

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Polymerase θ Coordinates Multiple Intrinsic Enzymatic Activities during DNA Repair

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Polθ reverse transcribes RNA and promotes RNA-templated DNA repair

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Jia Zhou et al.

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