DNA polymerase α-primase facilitates PARP inhibitor-induced fork acceleration and protects BRCA1-deficient cells against ssDNA gaps.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
27 Aug 2024
27 Aug 2024
Historique:
received:
27
10
2023
accepted:
14
08
2024
medline:
28
8
2024
pubmed:
28
8
2024
entrez:
27
8
2024
Statut:
epublish
Résumé
PARP inhibitors (PARPi), known for their ability to induce replication gaps and accelerate replication forks, have become potent agents in anticancer therapy. However, the molecular mechanism underlying PARPi-induced fork acceleration has remained elusive. Here, we show that the first PARPi-induced effect on DNA replication is an increased replication fork rate, followed by a secondary reduction in origin activity. Through the systematic knockdown of human DNA polymerases, we identify POLA1 as mediator of PARPi-induced fork acceleration. This acceleration depends on both DNA polymerase α and primase activities. Additionally, the depletion of POLA1 increases the accumulation of replication gaps induced by PARP inhibition, sensitizing cells to PARPi. BRCA1-depleted cells are especially susceptible to the formation of replication gaps under POLA1 inhibition. Accordingly, BRCA1 deficiency sensitizes cells to POLA1 inhibition. Thus, our findings establish the POLA complex as important player in PARPi-induced fork acceleration and provide evidence that lagging strand synthesis represents a targetable vulnerability in BRCA1-deficient cells.
Identifiants
pubmed: 39191785
doi: 10.1038/s41467-024-51667-1
pii: 10.1038/s41467-024-51667-1
doi:
Substances chimiques
Poly(ADP-ribose) Polymerase Inhibitors
0
DNA Primase
EC 2.7.7.-
BRCA1 Protein
0
DNA, Single-Stranded
0
BRCA1 protein, human
0
DNA polymerase alpha-primase
EC 2.7.7.-
DNA-Directed DNA Polymerase
EC 2.7.7.7
DNA Polymerase I
EC 2.7.7.7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7375Subventions
Organisme : Grantová Agentura České Republiky (Grant Agency of the Czech Republic)
ID : 20-03457Y
Organisme : Ministerstvo Školství, Mládeže a Tělovýchovy (Ministry of Education, Youth and Sports)
ID : LM2023050
Informations de copyright
© 2024. The Author(s).
Références
Conti, C. et al. Replication fork velocities at adjacent replication origins are coordinately modified during DNA replication in human cells. Mol. Biol. Cell 18, 3059–3067 (2007).
doi: 10.1091/mbc.e06-08-0689
pubmed: 17522385
pmcid: 1949372
Loeb, L. A. & Monnat, R. J. Jr. DNA polymerases and human disease. Nat. Rev. Genet. 9, 594–604 (2008).
doi: 10.1038/nrg2345
pubmed: 18626473
Lange, S. S., Takata, K. & Wood, R. D. DNA polymerases and cancer. Nat. Rev. Cancer 11, 96–110 (2011).
doi: 10.1038/nrc2998
pubmed: 21258395
pmcid: 3739438
Burgers, P. M. J. & Kunkel, T. A. Eukaryotic DNA replication fork. Annu. Rev. Biochem. 86, 417–438 (2017).
doi: 10.1146/annurev-biochem-061516-044709
pubmed: 28301743
pmcid: 5597965
Azarm, K. & Smith, S. Nuclear PARPs and genome integrity. Genes Dev. 34, 285–301 (2020).
doi: 10.1101/gad.334730.119
pubmed: 32029453
pmcid: 7050482
Ray Chaudhuri, A. & Nussenzweig, A. The multifaceted roles of PARP1 in DNA repair and chromatin remodelling. Nat. Rev. Mol. Cell Biol. 18, 610–621 (2017).
doi: 10.1038/nrm.2017.53
pubmed: 28676700
pmcid: 6591728
Rose, M., Burgess, J. T., O’Byrne, K., Richard, D. J. & Bolderson, E. PARP inhibitors: clinical relevance, mechanisms of action and tumor resistance. Front. Cell Dev. Biol. 8, 564601 (2020).
doi: 10.3389/fcell.2020.564601
pubmed: 33015058
pmcid: 7509090
Bryant, H. E. et al. Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 434, 913–917 (2005).
doi: 10.1038/nature03443
pubmed: 15829966
Farmer, H. et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 434, 917–921 (2005).
doi: 10.1038/nature03445
pubmed: 15829967
Maya-Mendoza, A. et al. High speed of fork progression induces DNA replication stress and genomic instability. Nature 559, 279–284 (2018).
doi: 10.1038/s41586-018-0261-5
pubmed: 29950726
Cong, K. et al. Replication gaps are a key determinant of PARP inhibitor synthetic lethality with BRCA deficiency. Mol. Cell 81, 3128–3144 e3127 (2021).
doi: 10.1016/j.molcel.2021.06.011
pubmed: 34216544
pmcid: 9089372
Cantor, S. B. Revisiting the BRCA-pathway through the lens of replication gap suppression: “Gaps determine therapy response in BRCA mutant cancer”. DNA Repair 107, 103209 (2021).
doi: 10.1016/j.dnarep.2021.103209
pubmed: 34419699
pmcid: 9049047
Vaitsiankova, A. et al. PARP inhibition impedes the maturation of nascent DNA strands during DNA replication. Nat. Struct. Mol. Biol. 29, 329–338 (2022).
Tirman, S., Cybulla, E., Quinet, A., Meroni, A. & Vindigni, A. PRIMPOL ready, set, reprime! Crit. Rev. Biochem. Mol. Biol. 56, 17–30 (2021).
doi: 10.1080/10409238.2020.1841089
pubmed: 33179522
Quinet, A. et al. PRIMPOL-mediated adaptive response suppresses replication fork reversal in BRCA-deficient cells. Mol. Cell 77, 461–474.e469 (2020).
doi: 10.1016/j.molcel.2019.10.008
pubmed: 31676232
pmcid: 7007862
Bai, G. et al. HLTF promotes fork reversal, limiting replication stress resistance and preventing multiple mechanisms of unrestrained DNA synthesis. Mol. Cell 78, 1237–1251.e1237 (2020).
doi: 10.1016/j.molcel.2020.04.031
pubmed: 32442397
pmcid: 7305998
Gonzalez-Acosta, D. et al. PrimPol-mediated repriming facilitates replication traverse of DNA interstrand crosslinks. EMBO J. 40, e106355 (2021).
doi: 10.15252/embj.2020106355
pubmed: 34128550
pmcid: 8280817
Piberger, A. L. et al. PrimPol-dependent single-stranded gap formation mediates homologous recombination at bulky DNA adducts. Nat. Commun. 11, 5863 (2020).
doi: 10.1038/s41467-020-19570-7
pubmed: 33203852
pmcid: 7673990
Rodriguez-Acebes, S., Mouron, S. & Mendez, J. Uncoupling fork speed and origin activity to identify the primary cause of replicative stress phenotypes. J. Biol. Chem. 293, 12855–12861 (2018).
doi: 10.1074/jbc.RA118.003740
pubmed: 29959228
pmcid: 6102153
Genois, M. M. et al. CARM1 regulates replication fork speed and stress response by stimulating PARP1. Mol. Cell 81, 784–800.e8 (2020).
Giansanti, C. et al. MDM2 binds and ubiquitinates PARP1 to enhance DNA replication fork progression. Cell Rep. 39, 110879 (2022).
doi: 10.1016/j.celrep.2022.110879
pubmed: 35649362
Han, T. et al. The antitumor toxin CD437 is a direct inhibitor of DNA polymerase alpha. Nat. Chem. Biol. 12, 511–515 (2016).
doi: 10.1038/nchembio.2082
pubmed: 27182663
pmcid: 4912453
Ercilla, A. et al. Physiological tolerance to ssDNA enables strand uncoupling during DNA replication. Cell Rep. 30, 2416–2429.e2417 (2020).
doi: 10.1016/j.celrep.2020.01.067
pubmed: 32075739
Holzer, S. et al. Structural basis for inhibition of human primase by arabinofuranosyl nucleoside analogues fludarabine and vidarabine. ACS Chem. Biol. 14, 1904–1912 (2019).
doi: 10.1021/acschembio.9b00367
pubmed: 31479243
pmcid: 6757278
Kang, Z. et al. BRCA2 associates with MCM10 to suppress PRIMPOL-mediated repriming and single-stranded gap formation after DNA damage. Nat. Commun. 12, 5966 (2021).
doi: 10.1038/s41467-021-26227-6
pubmed: 34645815
pmcid: 8514439
Panzarino, N. J. et al. Replication gaps underlie BRCA-deficiency and therapy response. Cancer Res. 81, 1388–1397 (2020).
Simoneau, A., Xiong, R. & Zou, L. The trans cell cycle effects of PARP inhibitors underlie their selectivity toward BRCA1/2-deficient cells. Genes Dev. 35, 1271–1289 (2021).
doi: 10.1101/gad.348479.121
pubmed: 34385259
pmcid: 8415318
Taglialatela, A. et al. REV1-Polzeta maintains the viability of homologous recombination-deficient cancer cells through mutagenic repair of PRIMPOL-dependent ssDNA gaps. Mol. Cell 81, 4008–4025.e7 (2021).
Tirman, S. et al. Temporally distinct post-replicative repair mechanisms fill PRIMPOL-dependent ssDNA gaps in human cells. Mol. Cell 81, 4026–4040.e4028 (2021).
doi: 10.1016/j.molcel.2021.09.013
pubmed: 34624216
pmcid: 8555837
Paes Dias, M. et al. Loss of nuclear DNA ligase III reverts PARP inhibitor resistance in BRCA1/53BP1 double-deficient cells by exposing ssDNA gaps. Mol. Cell 81, 4692–4708.e4699 (2021).
doi: 10.1016/j.molcel.2021.09.005
pubmed: 34555355
Kunkel, T. A., Hamatake, R. K., Motto-Fox, J., Fitzgerald, M. P. & Sugino, A. Fidelity of DNA polymerase I and the DNA polymerase I-DNA primase complex from Saccharomyces cerevisiae. Mol. Cell Biol. 9, 4447–4458 (1989).
pubmed: 2555694
pmcid: 362528
Zimmermann, M. et al. CRISPR screens identify genomic ribonucleotides as a source of PARP-trapping lesions. Nature 559, 285–289 (2018).
doi: 10.1038/s41586-018-0291-z
pubmed: 29973717
pmcid: 6071917
Nayak, S. et al. Inhibition of the translesion synthesis polymerase REV1 exploits replication gaps as a cancer vulnerability. Sci. Adv. 6, eaaz7808 (2020).
doi: 10.1126/sciadv.aaz7808
pubmed: 32577513
pmcid: 7286678
Thakar, T. et al. Lagging strand gap suppression connects BRCA-mediated fork protection to nucleosome assembly through PCNA-dependent CAF-1 recycling. Nat. Commun. 13, 5323 (2022).
doi: 10.1038/s41467-022-33028-y
pubmed: 36085347
pmcid: 9463168
Valli, C. et al. Atypical retinoids ST1926 and CD437 are S-phase-specific agents causing DNA double-strand breaks: significance for the cytotoxic and antiproliferative activity. Mol. Cancer Ther. 7, 2941–2954 (2008).
doi: 10.1158/1535-7163.MCT-08-0419
pubmed: 18790775
Johnson, N. et al. Stabilization of mutant BRCA1 protein confers PARP inhibitor and platinum resistance. Proc. Natl Acad. Sci. USA 110, 17041–17046 (2013).
doi: 10.1073/pnas.1305170110
pubmed: 24085845
pmcid: 3801063
Takata, K. et al. Conserved overlapping gene arrangement, restricted expression, and biochemical activities of DNA polymerase nu (POLN). J. Biol. Chem. 290, 24278–24293 (2015).
doi: 10.1074/jbc.M115.677419
pubmed: 26269593
pmcid: 4591814