Budd, M. E. & Campbell, J. L. A yeast gene required for DNA replication encodes a protein with homology to DNA helicases. Proc. Natl Acad. Sci. USA 92, 7642–7646 (1995).
Hu, J. et al. The intra-S phase checkpoint targets Dna2 to prevent stalled replication forks from reversing. Cell 149, 1221–1232 (2012).
Lin, W. et al. Mammalian DNA2 helicase/nuclease cleaves G-quadruplex DNA and is required for telomere integrity. EMBO J. 32, 1425–1439 (2013).
Karanja, K. K., Lee, E. H., Hendrickson, E. A. & Campbell, J. L. Preventing over-resection by DNA2 helicase/nuclease suppresses repair defects in Fanconi anemia cells. Cell Cycle 13, 1540–1550 (2014).
Shaheen, R. et al. Genomic analysis of primordial dwarfism reveals novel disease genes. Genome Res. 24, 291–299 (2014).
Tarnauskaitė, Ž. et al. Biallelic variants in DNA2 cause microcephalic primordial dwarfism. Hum. Mutat. 40, 1063–1070 (2019).
Di Lazzaro Filho, R. et al. Biallelic variants in DNA2 cause poikiloderma with congenital cataracts and severe growth failure reminiscent of Rothmund-Thomson syndrome. J. Med. Genet. 60, 1127–1132 (2023).
Peng, G. et al. Human nuclease/helicase DNA2 alleviates replication stress by promoting DNA end resection. Cancer Res. 72, 2802–2813 (2012).
Strauss, C. et al. The DNA2 nuclease/helicase is an estrogen-dependent gene mutated in breast and ovarian cancers. Oncotarget 5, 9396–9409 (2014).
Lu, Y. et al. Characteristic analysis of featured genes associated with stemness indices in colorectal cancer. Front. Mol. Biosci. 7, 563922 (2020).
Thongon, N. et al. Targeting DNA2 overcomes metabolic reprogramming in multiple myeloma. Nat. Commun. 15, 1203 (2024).
Hudson, J. J. R. & Rass, U. DNA2 in chromosome stability and cell survival—is it all about replication forks? Int. J. Mol. Sci. 22, 3984 (2021).
Burgers, P. M. It’s all about flaps: Dna2 and checkpoint activation. Cell Cycle 10, 2417–2418 (2011).
Bae, S. H., Bae, K. H., Kim, J. A. & Seo, Y. S. RPA governs endonuclease switching during processing of Okazaki fragments in eukaryotes. Nature 412, 456–461 (2001).
Zou, L. & Elledge, S. J. Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300, 1542–1548 (2003).
Budd, M. E., Antoshechkin, I. A., Reis, C., Wold, B. J. & Campbell, J. L. Inviability of a DNA2 deletion mutant is due to the DNA damage checkpoint. Cell Cycle 10, 1690–1698 (2011).
Duxin, J. P. et al. Okazaki fragment processing-independent role for human Dna2 enzyme during DNA replication. J. Biol. Chem. 287, 21980–21991 (2012).
Appanah, R., Jones, D., Falquet, B. & Rass, U. Limiting homologous recombination at stalled replication forks is essential for cell viability: DNA2 to the rescue. Curr. Genet. 66, 1085–1092 (2020).
Thangavel, S. et al. DNA2 drives processing and restart of reversed replication forks in human cells. J. Cell Biol. 208, 545–562 (2015).
Ölmezer, G. et al. Replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1. Nat. Commun. 7, 13157 (2016).
Rossi, S. E., Foiani, M. & Giannattasio, M. Dna2 processes behind the fork long ssDNA flaps generated by Pif1 and replication-dependent strand displacement. Nat. Commun. 9, 4830 (2018).
Falquet, B. et al. Disease-associated DNA2 nuclease-helicase protects cells from lethal chromosome under-replication. Nucleic Acids Res. 48, 7265–7278 (2020).
Liu, W. et al. FANCD2 and RAD51 recombinase directly inhibit DNA2 nuclease at stalled replication forks and FANCD2 acts as a novel RAD51 mediator in strand exchange to promote genome stability. Nucleic Acids Res. 51, 9144–9165 (2023).
Schlacher, K. et al. Double-strand break repair-independent role for BRCA2 in blocking stalled replication fork degradation by MRE11. Cell 145, 529–542 (2011).
Higgs, M. R. et al. BOD1L is required to suppress deleterious resection of stressed replication forks. Mol. Cell 59, 462–477 (2015).
Ahn, J. S., Osman, F. & Whitby, M. C. Replication fork blockage by RTS1 at an ectopic site promotes recombination in fission yeast. EMBO J. 24, 2011–2023 (2005).
Lambert, S., Watson, A., Sheedy, D. M., Martin, B. & Carr, A. M. Gross chromosomal rearrangements and elevated recombination at an inducible site-specific replication fork barrier. Cell 121, 689–702 (2005).
Petermann, E., Orta, M. L., Issaeva, N., Schultz, N. & Helleday, T. Hydroxyurea-stalled replication forks become progressively inactivated and require two different RAD51-mediated pathways for restart and repair. Mol. Cell 37, 492–502 (2010).
Ait Saada, A., Lambert, S. A. E. & Carr, A. M. Preserving replication fork integrity and competence via the homologous recombination pathway. DNA Repair 71, 135–147 (2018).
Teixeira-Silva, A. et al. The end-joining factor Ku acts in the end-resection of double strand break-free arrested replication forks. Nat. Commun. 8, 1982 (2017).
Carr, A. M. & Lambert, S. Replication stress-induced genome instability: the dark side of replication maintenance by homologous recombination. J. Mol. Biol. 425, 4733–4744 (2013).
Lydeard, J. R., Jain, S., Yamaguchi, M. & Haber, J. E. Break-induced replication and telomerase-independent telomere maintenance require Pol32. Nature 448, 820–823 (2007).
Costantino, L. et al. Break-induced replication repair of damaged forks induces genomic duplications in human cells. Science 343, 88–91 (2014).
Xu, Y. et al. DNA nicks in both leading and lagging strand templates can trigger break-induced replication. Mol. Cell 85, 91–106 (2025).
Tanaka, H., Ryu, G. H., Seo, Y. S. & MacNeill, S. A. Genetics of lagging strand DNA synthesis and maturation in fission yeast: suppression analysis links the Dna2-Cdc24 complex to DNA polymerase delta. Nucleic Acids Res. 32, 6367–6377 (2004).
Wu, X. & Malkova, A. Break-induced replication mechanisms in yeast and mammals. Curr. Opin. Genet. Dev. 71, 163–170 (2021).
Miyabe, I. et al. Polymerase delta replicates both strands after homologous recombination-dependent fork restart. Nat. Struct. Mol. Biol. 22, 932–938 (2015).
Pinter, S. F., Aubert, S. D. & Zakian, V. A. The Schizosaccharomyces pombe Pfh1p DNA helicase is essential for the maintenance of nuclear and mitochondrial DNA. Mol. Cell. Biol. 28, 6594–6608 (2008).
Li, Z. et al. hDNA2 nuclease/helicase promotes centromeric DNA replication and genome stability. EMBO J. 37, e96729 (2018).
Natsume, T., Kiyomitsu, T., Saga, Y. & Kanemaki, M. T. Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15, 210–218 (2016).
Chung, H. K. et al. Tunable and reversible drug control of protein production via a self-excising degron. Nat. Chem. Biol. 11, 713–720 (2015).
Hegarat, N. et al. Cyclin A triggers mitosis either via the Greatwall kinase pathway or cyclin B. EMBO J. 39, e104419 (2020).
Vaitsiankova, A. et al. PARP inhibition impedes the maturation of nascent DNA strands during DNA replication. Nat. Struct. Mol. Biol. 29, 329–338 (2022).
Hanzlikova, H. et al. The importance of poly(ADP-ribose) polymerase as a sensor of unligated Okazaki fragments during DNA replication. Mol. Cell 71, 319–331 (2018).
Ward, I. M. & Chen, J. Histone H2AX is phosphorylated in an ATR-dependent manner in response to replicational stress. J. Biol. Chem. 276, 47759–47762 (2001).
Fernandez-Casanas, M. & Chan, K. L. The unresolved problem of DNA bridging. Genes 9, 623 (2018).
Mocanu, C. et al. DNA replication is highly resilient and persistent under the challenge of mild replication stress. Cell Rep. 39, 110701 (2022).
Liu, W., Krishnamoorthy, A., Zhao, R. & Cortez, D. Two replication fork remodeling pathways generate nuclease substrates for distinct fork protection factors. Sci. Adv. 6, eabc3598 (2020).
Huang, F. et al. Identification of specific inhibitors of human RAD51 recombinase using high-throughput screening. ACS Chem. Biol. 6, 628–635 (2011).
Feringa, F. M. et al. Persistent repair intermediates induce senescence. Nat. Commun. 9, 3923 (2018).
Feringa, F. M. et al. Hypersensitivity to DNA damage in antephase as a safeguard for genome stability. Nat. Commun. 7, 12618 (2016).
Lossaint, G. et al. Reciprocal regulation of p21 and Chk1 controls the cyclin D1-RB pathway to mediate senescence onset after G2 arrest. J. Cell Sci. 135, jcs259114 (2022).
Müllers, E., Silva Cascales, H., Jaiswal, H., Saurin, A. T. & Lindqvist, A. Nuclear translocation of cyclin B1 marks the restriction point for terminal cell cycle exit in G2 phase. Cell Cycle 13, 2733–2743 (2014).
Fousek-Schuller, V. J. & Borgstahl, G. E. O. The intriguing mystery of RPA phosphorylation in DNA double-strand break repair. Genes 15, 167 (2024).
Minocherhomji, S. et al. Replication stress activates DNA repair synthesis in mitosis. Nature 528, 286–290 (2015).
Pinto, C., Kasaciunaite, K., Seidel, R. & Cejka, P. Human DNA2 possesses a cryptic DNA unwinding activity that functionally integrates with BLM or WRN helicases. eLife 5, e18574 (2016).
Formosa, T. & Nittis, T. Dna2 mutants reveal interactions with Dna polymerase alpha and Ctf4, a Pol alpha accessory factor, and show that full Dna2 helicase activity is not essential for growth. Genetics 151, 1459–1470 (1999).
d’Adda di Fagagna, F. Living on a break: cellular senescence as a DNA-damage response. Nat. Rev. Cancer 8, 512–522 (2008).
Macheret, M. & Halazonetis, T. D. DNA replication stress as a hallmark of cancer. Annu. Rev. Pathol. 10, 425–448 (2015).
Kumar, S. et al. Inhibition of DNA2 nuclease as a therapeutic strategy targeting replication stress in cancer cells. Oncogenesis 6, e319 (2017).
Liu, W. et al. A selective small molecule DNA2 inhibitor for sensitization of human cancer cells to chemotherapy. eBioMedicine 6, 73–86 (2016).
Folly-Kossi, H., Graves, J. D., Garan, L. A. W., Lin, F. T. & Lin, W. C. DNA2 nuclease inhibition confers synthetic lethality in cancers with mutant p53 and synergizes with PARP inhibitors. Cancer Res. Commun. 3, 2096–2112 (2023).
Klingseisen, A. & Jackson, A. P. Mechanisms and pathways of growth failure in primordial dwarfism. Genes Dev. 25, 2011–2024 (2011).
Martins, D. J., Di Lazzaro Filho, R., Bertola, D. R. & Hoch, N. C. Rothmund-Thomson syndrome, a disorder far from solved. Front. Aging 4, 1296409 (2023).
Nielsen-Dandoroff, E., Ruegg, M. S. G. & Bicknell, L. S. The expanding genetic and clinical landscape associated with Meier-Gorlin syndrome. Eur. J. Hum. Genet. 31, 859–868 (2023).
Moreno, S., Klar, A. & Nurse, P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 194, 795–823 (1991).
Watson, A. T., Garcia, V., Bone, N., Carr, A. M. & Armstrong, J. Gene tagging and gene replacement using recombinase-mediated cassette exchange in Schizosaccharomyces pombe. Gene 407, 63–74 (2008).
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
Osman, F. & Whitby, M. C. in DNA Replication: Methods and Protocols (eds Vengrova, S. & Dalgaard, J. Z.) 535–552 (Humana, 2009).
Keszthelyi, A., Daigaku, Y., Ptasinska, K., Miyabe, I. & Carr, A. M. Mapping ribonucleotides in genomic DNA and exploring replication dynamics by polymerase usage sequencing (Pu-seq). Nat. Protoc. 10, 1786–1801 (2015).
Naiman, K. et al. Replication dynamics of recombination-dependent replication forks. Nat. Commun. 12, 923 (2021).
Labun, K. et al. CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing. Nucleic Acids Res. 47, W171–W174 (2019).
Ran, F. A. et al. Genome engineering using the CRISPR-Cas9 system. Nat. Protoc. 8, 2281–2308 (2013).
Kowarz, E., Loscher, D. & Marschalek, R. Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol. J. 10, 647–653 (2015).
