RHINO directs MMEJ to repair DNA breaks in mitosis

Science, Journal Year: 2023, Volume and Issue: 381(6658), P. 653 - 660

Published: July 13, 2023

Nonhomologous end-joining (NHEJ) and homologous recombination (HR) are the primary pathways for repairing DNA double-strand breaks (DSBs) during interphase, whereas microhomology-mediated (MMEJ) has been regarded as a backup mechanism. Through CRISPR-Cas9-based synthetic lethal screens in cancer cells, we identified subunits of 9-1-1 complex (RAD9A-RAD1-HUS1) its interacting partner, RHINO, crucial MMEJ factors. We uncovered an unexpected function RHINO restricting to mitosis. accumulates M phase, undergoes Polo-like kinase 1 (PLK1) phosphorylation, interacts with polymerase θ (Polθ), enabling recruitment DSBs subsequent repair. Additionally, provide evidence that activity mitosis repairs persistent originate S phase. Our findings offer insights into relationship between genes

Language: Английский

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Revolutionizing DNA repair research and cancer therapy with CRISPR–Cas screens

Nature Reviews Molecular Cell Biology, Journal Year: 2023, Volume and Issue: 24(7), P. 477 - 494

Published: Feb. 13, 2023

Language: Английский

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Base Excision Repair: Mechanisms and Impact in Biology, Disease, and Medicine

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(18), P. 14186 - 14186

Published: Sept. 16, 2023

Base excision repair (BER) corrects forms of oxidative, deamination, alkylation, and abasic single-base damage that appear to have minimal effects on the helix. Since its discovery in 1974, field has grown several facets: mechanisms, biology physiology, understanding deficiencies human disease, using BER genes as potential inhibitory targets develop therapeutics. Within segregation short nucleotide (SN-) long patch (LP-), there are currently six known global with emerging work transcription- replication-associated BER. Knockouts (KOs) mouse models showed single glycosylase knockout had phenotypic impact, but were clearly seen double knockouts. However, KOs downstream enzymes critical impact health survival mice. gene deficiency contributes cancer, inflammation, aging, neurodegenerative disorders. Medicinal being developed for or combinatorial therapies, only PARP APE1 yet reach clinical stage.

Language: Английский

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EXO1 protects BRCA1-deficient cells against toxic DNA lesions

Molecular Cell, Journal Year: 2024, Volume and Issue: 84(4), P. 659 - 674.e7

Published: Jan. 23, 2024

Inactivating mutations in the BRCA1 and BRCA2 genes impair DNA double-strand break (DSB) repair by homologous recombination (HR), leading to chromosomal instability cancer. Importantly, BRCA1/2 deficiency also causes therapeutically targetable vulnerabilities. Here, we identify dependency on end resection factor EXO1 as a key vulnerability of BRCA1-deficient cells. generates poly(ADP-ribose)-decorated lesions during S phase that associate with unresolved DSBs genomic but not wild-type or BRCA2-deficient Our data indicate BRCA1/EXO1 double-deficient cells accumulate due impaired single-strand annealing (SSA) top their HR defect. In contrast, retain SSA activity absence hence tolerate loss. Consistent EXO1-mediated SSA, find BRCA1-mutated tumors show elevated expression increased SSA-associated scars compared BRCA1-proficient tumors. Overall, our findings uncover promising therapeutic target for

Language: Английский

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Targeting the DNA Repair Enzyme Polymerase θ in Cancer Therapy

Trends in cancer, Journal Year: 2020, Volume and Issue: 7(2), P. 98 - 111

Published: Oct. 24, 2020

POLQ is a versatile DNA repair enzyme that central in TMEJ for the error-prone of DSBs. also functions other pathways including base excision repair, interstrand crosslink and damage tolerance by translesion synthesis.Cancer cells often acquire mutations genes, making them dependent on remaining pathways. Dependence characterized an increased expression which associated with poor patient prognosis.Depletion POLQ-dependent cancers leads to synthetic lethality. This well described malignancies deficient homologous recombination (e.g., due BRCA1 or BRCA2). Hence, use inhibitors might be promising strategy targeted cancer therapy. Targeted therapies represent milestone towards personalized treatment as they function via inhibition cancer-specific alterations. Polymerase θ (POLQ), polymerase, involved double-strand break (DSB) upregulated cancer. lethal various known drivers such BRCA1/2, it essential recombination-deficient cancers. Thus, represents target therapy efforts development are actively underway first clinical trials start 2021. review summarizes journey from backup therapeutic treatment. To increase efficiency lower burden toxic side effects, major goal progress ‘one-drug-fits-all’ individualized approach tailored tumor-specific molecular features. Two main strategies currently utilized treatment, both exploiting vulnerabilities. In approach, suppression aberrantly oncogenes alleviates growth advantage cells. The second based phenomenon genetic alterations acquired tumor cause their dependency compensatory pathways, loss lethality (see Glossary). Therefore, alteration evokes cellular death while leaving normal unharmed [1.Dobzhansky T. Genetics natural populations; variability populations Drosophila pseudoobscura.Genetics. 1946; 31: 269-290Crossref PubMed Google Scholar]. recent advent genome-wide interaction studies has demonstrated extensive number interactions cancer, many can potentially translated [2.Han K. et al.Synergistic drug combinations identified CRISPR screen pairwise interactions.Nat. Biotechnol. 2017; 35: 463-474Crossref Scopus (250) Cancer frequently genes respond rewiring network utilize survival. Dependency opens room small molecule inhibitors. A group successful drugs this mode action poly(ADP-ribose) polymerase (PARP) inhibitors, approved BRCA-deficient essentiality PARP loss-of-function BRCA1/2 remarkable up 1000 times more sensitive than healthy [3.Farmer H. al.Targeting defect BRCA mutant strategy.Nature. 2005; 434: 917-921Crossref (4441) Scholar,4.Bryant H.E. al.Specific killing BRCA2-deficient tumours polymerase.[erratum appears Nature. 2007 May 17;447(7142):346].Nature. 913-917Crossref (3464) Although challenges acquisition resistance need faced, success inhibitory targeting enzymes highly encouraging. context, DNA-repair (POLQ) received increasing attention. numerous its overexpression prognosis [5.Kawamura al.DNA preferentially expressed lymphoid tissues human cancers.Int. J. Cancer. 2004; 109: 9-16Crossref (94) Scholar, 6.Lemée F. up-regulation survival breast perturbs replication, promotes instability.Proc. Natl. Acad. Sci. U. S. A. 2010; 107: 13390-13395Crossref (115) 7.Higgins G.S. al.Overexpression confers early patients.Oncotarget. 1: 175-184Crossref (71) 8.Lessa R. al.Adult height head neck cancer: pooled analysis within INHANCE Consortium.Head Neck. 2014; 36: 1391Google 9.Allera-Moreau C. replication stress response involving PLK1, CDC6, POLQ, RAD51 CLASPIN upregulation prognoses outcome early/mid-stage non-small cell lung patients.Oncogenesis. 2012; 1-10Crossref (68) Moreover, between multiple factors (such BRCA1/2), have been [10.Shima N. al.The mouse genomic instability mutation chaos1 allele Polq exhibits Atm.Mol. Cell. Biol. 24: 10381-10389Crossref (105) 11.Wang Z. important breaks caused fork collapse.J. Chem. 2019; 294: 3909-3919Abstract Full Text PDF (43) 12.Ceccaldi al.Homologous-recombination-deficient Polθ -mediated repair.Nature. 2015; 518: 258-262Crossref (445) 13.Mateos-Gomez P.A. al.Mammalian alternative NHEJ suppresses recombination.Nature. 254-257Crossref (378) 14.Kelso A.A. al.Distinct roles RAD52 chromosomal response.PLoS Genet. 15: 1-28Crossref (36) 15.Feng W. al.Genetic determinants addiction theta.Nat. Commun. (Published online September 19, 2019. https://doi.org/10.1038/s41467-019-12234-1)Crossref (57) 16.Mengwasser K.E. Screens Reveal FEN1 APEX2 BRCA2 Synthetic Lethal Targets.Mol. 73 (e6): 885-899Abstract (83) For these reasons, biotech companies laboratories, soon tested trials. review, we focus unique protein structure allows fulfill diverse roles. We further discuss conflicting evidence whether stability, given intrinsically synthesis enzyme. Finally, address why meets criteria summarize state-of-the art inhibitor development. (DSBs), most cytotoxic type lesion. If unrepaired, DSBs deleterious consequences rearrangements death. specialized consisting at least three responsible (Figure 1A ). Most repaired canonical nonhomologous end joining (c-NHEJ), pathway directly religates ends without processing, introducing insertions deletions sites [17.Chang H.H.Y. al.Non-homologous repair.Nat. Rev. Mol. Cell 18: 495-506Crossref (706) S G2 phases cycle, when sister chromatid available, (HR) favored only precise DSB [18.Ranjha L. al.Main steps repair: introduction related processes.Chromosoma. 2018; 127: 187-214Crossref (148) third [originally named (alt-EJ) microhomology-mediated (MMEJ)] was later termed theta-mediated (TMEJ) requirement [19.Koole al.A theta-dependent endogenous G4 sites.Nat. 5: (140) initiated PARP1 recruitment resected DNA-ends [20.Audebert M. al.Involvement polymerase-1 XRCC1/DNA ligase III route rejoining.J. 279: 55117-55126Abstract (533) 21.Wang al.PARP-1 Ku compete double strand distinct pathways.Nucleic Acids Res. 2006; 34: 6170-6182Crossref (572) 22.Truong L.N. al.Microhomology-mediated share initial resection step mammalian cells.Proc. 2013; 110: 7720-7725Crossref (282) Upon activation phosphorylated CtIP, 3′ overhangs generated helicases MRE11–RAD50–NBS1 (MRN) complex. then binds long single-stranded (ssDNA) 5′–3′ anneals sequences 2–6 pairs microhomology primers [23.Chan S.H. al.Dual theta end-joining Drosophila.PLoS 6: 1-16Crossref (296) 24.Kent al.Mechanism promoted θ.Nat. Struct. 22: 230-237Crossref (174) 25.Ahrabi role suppressing joining.Nucleic 2016; 44: 5743-5757Crossref (54) stabilized ligated LIG3–XRCC1 LIG1 [26.Simsek D. during translocation formation.PLoS 2011; 7: 1-11Crossref (213) 27.Liang al.Human ligases I III, but not IV, required breaks.Nucleic 2008; 3297-3310Crossref (109) 28.Lu G. al.Ligase mediate ligation end-joining.Proc. 113: 1256-1260Crossref (50) Scholar] 1A). Repair error prone introduces characteristic sequence alterations, called mutational signatures two attributes. Firstly, since uses microhomologies annealing yet minority contain regions, necessary make accessible. microhomologous may result microhomology-flanked [29.Sallmyr Tomkinson A.E. pathways.J. 293: 10536-10549Abstract (111) Secondly, tends abort template-dependent extension annealed reanneal secondary sequences. results short stretches de novo resembles flanking break, templated [30.Schimmel al.Templated Insertions: smoking gun joining.Trends 632-644Abstract (48) Templated originate opposite (in trans) same cis), protruding ssDNA snaps back itself [24.Kent Scholar,31.Kent al.Polymerase robust terminal transferase oscillates different mechanisms end-joining.Elife. 1-25Crossref (47) interestingly, map activity doing so, likely contributes variety loci mutated disorders, emphasizing TMEJ’s etiology diseases tightly regulated. G1 phase HR unavailable, association abundant Ku-heterodimer 53BP1 free inhibits resection, thereby channeling c-NHEJ [32.Bunting S.F. al.53BP1 brca1-deficient blocking breaks.Cell. 141: 243-254Abstract (1121) however, removed CtIP complex MRN, shifting balance favor HR. Since require ends, each substrate. displace RAD51, key factor, proposed RAD51-binding domain [12.Ceccaldi counteract RPA, another factor [33.Mateos-Gomez helicase counteracts RPA promote alt-NHEJ.Nat. 1116-1123Crossref (75) Furthermore, depletion proteins BRCA1, BRCA2, increases TMEJ-specific signature suggesting negatively regulated [25.Ahrabi Notably, high mutagenicity, considered merely pathway. However, becoming increasingly evident presence available specific types lesions [22.Truong Such include collapsed forks chromatids containing replication-obstructing crosslink) rendering unsuitable template [34.Wyatt D.W. al.Essential θ-mediated chromosome breaks.Mol. 63: 662-673Abstract (136) 1B). Caenorhabditis elegans, shown indispensable quadruplex structures, preventing expense Future research needed assess regulation order identify conditions particular lesions, depend activity. Increasing suggests thus incorporates nucleotides apurinic/apyrimidinic sites, thymine glycols, thymidine dimers [35.Seki al.POLQ (Pol θ), DNA-dependent ATPase cells.Nucleic 2003; 6117-6126Crossref (144) 36.Hogg al.Crystallographic snapshots replicative encountering abasic site.EMBO 23: 1483-1493Crossref (126) 37.Hogg al.Lesion bypass intrinsic property pol depends inserts.J. 405: 642-652Crossref (60) 38.Kusumoto al.Translesion η across glycol lesions.Biochemistry. 2002; 41: 6090-6099Crossref (118) 39.Takata K.I. N (POLN) low fidelity capable error-free 5S-thymine glycol.J. 281: 23445-23455Abstract 40.Yoon J.H. al.Error-prone through UV Lesions protects against skin cancers.Cell. 176: 1295-1309.e15Abstract addition, replication-associated [41.Alexander J.L. al.Multiple contribute rereplication follicle 13809-13814Crossref (16) Scholar,42.Roerink elegans.Genome 954-962Crossref Depletion decreased velocity amount stalled upon hydroxyurea, chemical used induce stalling single-strand converted into [11.Wang crosslinks (ICLs) Drosophila, Arabidopsis, elegans [43.Beagan al.Drosophila utilizes helicase-like domains repair.PLoS 13: 1-19Crossref (28) 44.Muzzini D.M. al.Caenorhabditis POLQ-1 HEL-308 cross-link pathways.DNA (Amst). 941-950Crossref (73) 45.Inagaki al.Arabidopsis TEBICHI , domains, division differentiation meristems.18. 2006: 879-892Google Scholar], systems demonstrate redundancy TLS polymerases Scholar,46.Yousefzadeh M.J. POLQ.PLoS 10e1004654Crossref (154) few exceptions reported: knockout (KO) embryonic fibroblasts (MEFs) hypersensitive mitomycin C, ICL-inducing agent, higher levels micronuclei C were observed mice [15.Feng Scholar,47.Shima al.Phenotype-based identification mutants.Genetics. 163: 1031-1040Crossref weak 5′-deoxyribose phosphate lyase domain, suggested act (BER) [48.Prasad possesses 5′-dRP single-nucleotide vitro.Nucleic 2009; 37: 1868-1877Crossref (78) although extent involvement matter debate [49.Yoshimura al.Vertebrate POLβ cooperate oxidative damage.Mol. 115-125Abstract 50.Ukai al.Role exogenous B cells.Genes Cells. 11: 111-121Crossref (26) 51.Arana M.E. al.Low-fidelity theta.Nucleic 3847-3856Crossref (106) 52.Higgins interfering RNA identifies radiosensitization knockdown.Cancer 70: 2984-2993Crossref (84) conclusion, deeper insights variation model organisms mechanistic lacking. encodes A-family contains N-terminal conserved superfamily 2 C-terminal linked unstructured region 2). As such, eukaryotic date domain. coordinated interplay all allow execution [53.Black S.J. al.Molecular basis purified full-length Polθ.Nat. 10 27, https://doi.org/10.1038/s41467-019-12272-9)Crossref either using Despite conservation, regulating substrate selection. version lacking perform substrates (≤26 nucleotides) whereas cannot motifs ATPase-dependent manner Scholar,35.Seki Scholar,54.Büttner al.Structural duplex separation superfamily-2 helicase.Nat. 2007; 14: 647-652Crossref (242) 55.Richards J.D. al.Structure Hel308 reveals binding autoinhibitory domains.J. 283: 5118-5126Abstract 56.Newman J.A. possible pathway.Structure. 2319-2330Abstract (46) performing longer alone unproductive snap-back mechanism summary, competing extension, connected flexible region. Whether debate. Biochemical culminates large Scholar,30.Schimmel Scholar,57.Seki al.High-efficiency Q.EMBO 4484-4494Crossref (161) Beyond vitro systems, performed yielded findings supporting opposing guardian stability (Table 1). protect stability: formation, exacerbates sensitivity genotoxic agents, destabilizes Scholar,12.Ceccaldi Scholar,14.Kelso Scholar,52.Higgins Scholar,58.Goff J.P. al.Lack radiosensitizes bone marrow stromal reticulocyte after total-body irradiation.Radiat. 172: 165-174Crossref Other studies, reported decreases translocations UV-associated markers impairs cycle progression [6.Lemée Scholar,13.Mateos-Gomez Scholar,40.Yoon cancers.Cel

Language: Английский

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Advances and perspectives of PARP inhibitors

Experimental Hematology and Oncology, Journal Year: 2019, Volume and Issue: 8(1)

Published: Nov. 11, 2019

DNA damage repair deficiency leads to the increased risk of genome instability and oncogenic transformation. In meanwhile, this could be exploited for cancer treatment by inducing excessive catastrophic damage. Continuous replication in cells higher demand components. Due loss some effectors (e.g. BRCA) incomplete repertoire, are addicted certain pathways such as Poly (ADP-ribose) polymerase (PARP)-related single-strand break pathway. The interaction between BRCA PARP is a form synthetic lethal effect which means simultaneously functional two genes lead cell death, while defect any single gene has slight on viability. Based theory, inhibitor (PARPi) was developed aiming selectively target harboring BRCA1/2 mutations. Recently, growing body evidence indicated that broader population patients benefit from PARPi therapy far beyond those with germline mutated tumors. Numerous biomarkers including homologous recombination high level pressure also herald sensitivity treatment. Besides, series studies PARPi-involved combination additional chemotherapy therapy, immune checkpoint inhibitor, well targeted agent had great advantage overcoming resistance enhancing efficacy. review, we summarized advances clinical application. highlighted multiple promising PARPi-based strategies preclinical studies.

Language: Английский

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A consensus set of genetic vulnerabilities to ATR inhibition

Open Biology, Journal Year: 2019, Volume and Issue: 9(9), P. 190156 - 190156

Published: Sept. 1, 2019

The response to DNA replication stress in eukaryotes is under the control of ataxia–telangiectasia and Rad3-related (ATR) kinase. ATR responds single-stranded (ss) stabilize distressed forks, modulate firing prevent cells with damaged or incomplete from entering into mitosis. Furthermore, inhibitors are currently clinical development either as monotherapies combination agents that perturb replication. To gain a genetic view cellular pathways requiring kinase function, we mapped genes whose mutation causes hypersensitivity genome-scale CRISPR/Cas9 screens. We delineate consensus set 117 enriched replication, repair cell cycle regulators promote survival when activity suppressed. validate 14 this report not previously described inhibitors. In particular found loss POLE3/POLE4 proteins, which polymerase ε accessory subunits, results marked inhibition. anticipate 117-gene will be useful for identification involved regulation genome integrity characterization new biological processes involving ATR, may reveal biomarkers inhibitor clinic.

Language: Английский

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Targeting the nucleotide salvage factor DNPH1 sensitizes BRCA -deficient cells to PARP inhibitors

Science, Journal Year: 2021, Volume and Issue: 372(6538), P. 156 - 165

Published: April 9, 2021

Three strikes to knock cancer out BRCA1 and BRCA2 are tumor-suppressor genes, patients with mutations in these genes predisposed breast, ovarian, other cancers. Because affect pathways involved DNA break repair, patients' tumors usually vulnerable treatments that further damage such as poly(ADP-ribose) polymerase (PARP) inhibitors, but they can acquire resistance therapy. Using a genome-wide screening approach, Fugger et al. identified protein called DNPH1 “nucleotide sanitizer” prevents the incorporation of abnormal nucleotides into (see Perspective by Kriaucionis). The authors examined its mechanism action demonstrated how it be targeted expedite killing -mutant cells combination PARP inhibitor treatment. Science , this issue p. 156 ; see also 127

Language: Английский

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Endogenous DNA 3′ Blocks Are Vulnerabilities for BRCA1 and BRCA2 Deficiency and Are Reversed by the APE2 Nuclease

Molecular Cell, Journal Year: 2020, Volume and Issue: 78(6), P. 1152 - 1165.e8

Published: June 1, 2020

Language: Английский

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Base excision repair and its implications to cancer therapy

Essays in Biochemistry, Journal Year: 2020, Volume and Issue: 64(5), P. 831 - 843

Published: July 10, 2020

Base excision repair (BER) has evolved to preserve the integrity of DNA following cellular oxidative stress and in response exogenous insults. The pathway is a coordinated, sequential process involving 30 proteins or more which single strand breaks are generated as intermediates during process. While deficiencies BER activity can lead high mutation rates tumorigenesis, cancer cells often rely on increased tolerate stress. Targeting been an attractive strategy overwhelm with damage, improve efficacy radiotherapy and/or chemotherapy, form part lethal combination specific mutation/loss function. We provide update progress inhibitors enzymes involved BER, some challenges faced targeting pathway.

Language: Английский

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