The trans cell cycle effects of PARP inhibitors underlie their selectivity toward BRCA1/2-deficient cells

Genes & Development, Journal Year: 2021, Volume and Issue: 35(17-18), P. 1271 - 1289

Published: Aug. 12, 2021

PARP inhibitor (PARPi) is widely used to treat BRCA1/2-deficient tumors, but why PARPi more effective than other DNA-damaging drugs unclear. Here, we show that generates DNA double-strand breaks (DSBs) predominantly in a trans cell cycle manner. During the first S phase after exposure, induces single-stranded (ssDNA) gaps behind replication forks. By trapping on DNA, prevents completion of gap repair until next phase, leading collisions forks with ssDNA and surge DSBs. In second cells are unable suppress origin firing through ATR, resulting continuous synthesis Furthermore, cannot recruit RAD51 collapsed Thus, DSBs progressively gaps, fail slow down over multiple cycles, explaining unique efficacy cells.

Language: Английский

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PARP1 proximity proteomics reveals interaction partners at stressed replication forks

Nucleic Acids Research, Journal Year: 2022, Volume and Issue: 50(20), P. 11600 - 11618

Published: Oct. 11, 2022

Abstract PARP1 mediates poly-ADP-ribosylation of proteins on chromatin in response to different types DNA lesions. PARP inhibitors are used for the treatment BRCA1/2-deficient breast, ovarian, and prostate cancer. Loss replication fork protection is proposed as one mechanism that contributes vulnerability cells inhibitors. However, mechanisms regulate activity at stressed forks remain poorly understood. Here, we performed proximity proteomics isolation map putative regulators. We identified TPX2 a direct PARP1-binding protein regulates auto-ADP-ribosylation PARP1. interacts with damage promotes homology-directed repair double-strand breaks. Moreover, mRNA levels increased BRCA1/2-mutated breast cancers, high expression correlate sensitivity cancer PARP-trapping propose confers mitosis-independent function cellular stress by interacting

Language: Английский

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ADP-ribose contributions to genome stability and PARP enzyme trapping on sites of DNA damage; paradigm shifts for a coming-of-age modification

Journal of Biological Chemistry, Journal Year: 2023, Volume and Issue: 299(12), P. 105397 - 105397

Published: Oct. 28, 2023

ADP-ribose is a versatile modification that plays critical role in diverse cellular processes. The addition of this catalyzed by ADP-ribosyltransferases, among which notable poly(ADP-ribose) polymerase (PARP) enzymes are intimately involved the maintenance genome integrity. modifications during DNA damage repair significant interest for proper development PARP inhibitors targeted toward treatment diseases caused genomic instability. More specifically, promoting persistence on lesions, termed "trapping," considered desirable characteristic. In review, we discuss key classes proteins signaling (writers, readers, and erasers) with focus those An overview factors modulate PARP1 PARP2 at sites lesions also discussed. Finally, clarify aspects trapping model light recent studies characterize kinetics recruitment lesions. These findings suggest could be as continuous molecules to rather than physical stalling molecules. Recent novel research tools have elevated level understanding ADP-ribosylation, marking coming-of-age interesting modification. carries necessary information many processes within cell maintaining its stability importance ensure viability. Genome instability can arise from endogenous causes, such normal transactions (replication, transcription, recombination), but exogenous like external damaging agents (1Chatterjee N. Walker G.C. Mechanisms damage, repair, mutagenesis.Environ. Mol. Mutagen. 2017; 58: 235-263Crossref PubMed Scopus (957) Google Scholar). sheer number each human experiences daily (approximately 70,000 lesions) (2Lindahl T. Barnes D.E. Repair damage.Cold Spring Harb. Symp. Quant. Biol. 2000; 65: 127-133Crossref Scholar) highlights heavy demand put mechanisms. As such, variety pathways exist tackle diversity abundance these carrying overlapping functions rely interplay between posttranslational (PTMs) (phosphorylation, ubiquitylation, SUMOylation, etc) proceed success (3Huen M.S. Chen J. response pathways: crossroad protein modifications.Cell Res. 2008; 18: 8-16Crossref (162) an ancient nucleic acid has been utilized organisms, often defense mechanism (4Lüscher B. Bütepage M. Eckei L. Krieg S. Verheugd P. Shilton B.H. multifaceted control physiology health disease.Chem. Rev. 2018; 118: 1092-1136Crossref (154) Mammalian cells employ contexts, including antiviral defense/innate immunity, homeostasis, gene regulation, repair/genome (5Luscher Ahel I. Altmeyer Ashworth A. Bai Chang et al.ADP-ribosyltransferases, update function nomenclature.FEBS 2021; 289: 7399-7410Crossref (104) Notably, single (ADPr) unit modifications, multiple ADPr joined polymer known or PAR. PAR chains linearly elongated through formation (2′-1″) ribose–ribose glycosidic bond units. Occasionally, (2″-1″) occur branches (Fig. 1A) (6Chen Q. Kassab M.A. Dantzer F. Yu X. mediates branched poly ADP-ribosylation damage.Nat. Commun. 9: 3233Crossref (97) Scholar, 7Alemasova E.E. Lavrik O.I. Poly(ADP-ribosyl)ation PARP1: reaction regulatory proteins.Nucleic Acids 2019; 47: 3811-3827Crossref (232) Although majority published investigated proteins, there growing evidence appreciation prevalence acids (8Musheev M.U. Schomacher Basu Han D. Krebs Scholz C. al.Mammalian N1-adenosine PARylation reversible modification.Nat. 2022; 13: 6138Crossref (9) 9Schuller Beyond modification: rise non-canonical ADP-ribosylation.Biochem. 479: 463-477Crossref (16) 10Weixler Scharinger K. Momoh Luscher Feijs K.L.H. Zaja R. RNA DNA: vitro characterization vivo function.Nucleic 49: 3634-3650Crossref (40) This review our current employed catalysis, turnover, signaling, enzymes. (PARPi) important biology several PARPi approved use cancer treatments. covers knowledge mode action, particular clarifying enigmatic process "trapping." ADP-ribosyltransferase (ART) take group NAD+ attach it macromolecules. Proteins modified amino sidechains, Glu, Asp, Ser, Arg, Cys Nucleic receive phosphorylated termini nucleobases diphtheria toxin-like family, containing mammalian enzymes, defined H-Y-[E/D/Q] signature motif their binding 1B). active site composed "donor" split into nicotinamide pocket, catalytic triad located, adenine pocket (7Alemasova effectively holds moiety will attached either target protein/nucleic chain undergoing elongation. elongation requires presence "acceptor" moiety, already target, new added most members family do not catalyze PARylation, they possess sites. include PARP1, PARP2, TNKS1 (PARP5a), TNKS2 (PARP5b) 1C). PARP3 participates catalyzes ADPr, mono-ADP-ribosylation (MARylation). A later section some mechanisms regulating writers specific roles maintenance. readers comprised modules recognize MAR without removing Many recruited via Among high-affinity PAR-binding (11Gagné J.P. Isabelle Lo K.S. Bourassa Hendzel M.J. Dawson V.L. al.Proteome-wide identification poly(ADP-ribose)-associated complexes.Nucleic 36: 6959-6976Crossref (320) zinc fingers (PBZs) (12Ahel Matsusaka Clark A.J. Pines Boulton S.J. al.Poly(ADP-ribose)-binding finger motifs repair/checkpoint proteins.Nature. 451: 81-85Crossref (332) For example, while p53 (a transcription activator) XPA scaffolding nucleotide excision repair) bind conserved (13Reber J.M. Mangerich Why structure length matter: biological significance underlying structural heterogeneity poly(ADP-ribose).Nucleic 8432-8448Crossref (0) Scholar), histone chaperone aprataxin polynucleotide kinase factor (APLF) two PBZ tandem APLF were found branching although currently unclear how may coordinate mediate (14Eustermann Brockmann Mehrotra P.V. Yang J.C. Loakes West S.C. al.Solution structures domains interaction poly(ADP-ribose).Nat. Struct. 2010; 17: 241-243Crossref (83) fact, preference reproduced study (15Löffler Krüger Zirak Winterhalder Müller A.L. Fischbach al.Influence poly(ADP-ribose)-protein interactions.Nucleic 2023; 51: 536-552Crossref (2) generally accepted low abundance, explain difficulty identifying specifically recognizing Other WWE BRCT 1D) Of note, RNA- DNA-recognition motifs, oligonucleotide/oligosaccharide-binding fold, interact essentially chemically similar DNA. shift PAR, RNA, DNA, depending (DDR) further discussed below. Enzymes digest remove referred erasers. Notable erasers glycohydrolase (PARG) (ADP-ribosyl)hydrolase 3 (ARH3) 1E). thorough reviews recently written about PARG, ARH3 structure, substrate recognition, (16Rack J.G.M. Liu Zorzini V. Voorneveld Ariza Honarmand Ebrahimi al.Mechanistic insights three steps poly(ADP-ribosylation) reversal.Nat. 12: 4581Crossref (33) 17Schützenhofer Rack making breaking serine-ADP-ribosylation response.Front. Cell Dev. 9745922Crossref (8) We provide summary activities section. PARG hydrolyzes high efficacy bonds chains. degrades linear chains, cannot last, protein-linked thus leaving MARylation mark targets (18Hatakeyama Nemoto Y. Ueda Hayaishi O. Purification glycohydrolase. Different modes action large small poly(ADP-ribose).J. Chem. 1986; 261: 14902-14911Abstract Full Text PDF 19Braun S.A. Panzeter P.L. Collinge Althaus F.R. Endoglycosidic cleavage polymers glycohydrolase.Eur. Biochem. 1994; 220: 369-375Crossref 20Barkauskaite E. Brassington Tan E.S. Warwicker Dunstan Banos al.Visualization bound reveals inherent balance exo- endo-glycohydrolase activities.Nat. 2013; 4: 2164Crossref (109) Interestingly, acts both exo-glycohydrolase (degrading starting terminus, releasing units) (21Slade Barkauskaite Weston Lafite Dixon al.The glycohydrolase.Nature. 2011; 477: 616-620Crossref (275) weak releases fragments (longer subsequently degraded itself, albeit inefficiently (20Barkauskaite 22Pourfarjam Kasson Tran Ho Lim Kim I.K. robust activity protein-free chains.Biochem. Biophys. 2020; 527: 818-823Crossref (13) removal left mono-ADP-ribosyl-acceptor hydrolases. one hydrolase acting DDR removes serine-linked forms (23Fontana Bonfiglio J.J. Palazzo Bartlett Matic Serine reversal ARH3.Elife. 6e28533Crossref (149) Erasers capable Glu/Asp residues typically macrodomains, MacroD1, MacroD2, terminal 1 (24Barkauskaite Jankevicius G. Structures synthesis degradation PARP-dependent ADP-ribosylation.Mol. Cell. 2015; 935-946Abstract (190) acids. phosphate-linked reversed 1, (9Schuller adenine-linked removed There still much work establish However, elucidated regulated strand breaks, potent stimulator production cells. Indeed, abundant enzyme primary writer cell, output accounts approximately 80 90% produced (25D'Amours Desnoyers D'Silva Poirier G.G. reactions regulation nuclear functions.Biochem. 1999; 342: 249-268Crossref (1612) domain architecture six independently folded domains: (Zn1, Zn2, Zn3), WGR (Trp-Gly-Arg) domain, (CAT) domain. CAT helical (HD) ART located localizes nucleus where scans intact chromatin intrastrand transfer, monkey-bar (26Rudolph Mahadevan Dyer Luger Poly(ADP-ribose) searches 'monkey bar' mechanism.Elife. 7e37818Crossref (42) transfer cooperative fingers, move molecule another 27Rudolph Muthurajan U.M. Palacio Roberts Erbse A.H. binds transfer.Mol. 81: 4994-5006.e5Abstract scanning does trigger (27Rudolph 28Benjamin R.C. Gill D.M. programmed damaged comparison different types breaks.J. 1980; 255: 10502-10508Abstract Rather, activated following efficient organization (29Langelier M.F. Planck J.L. Roy Pascal Structural basis damage-dependent poly(ADP-ribosyl)ation PARP-1.Science. 2012; 336: 728-732Crossref (465) 30Eustermann Wu W.F. Langelier Easton L.E. Riccio A.A. al.Structural detection single-strand breaks PARP-1.Mol. 60: 742-754Abstract (202) 31Rudolph Probing conformational changes associated PARP1.Biochemistry. 59: 2003-2011Crossref relays activating signal allosteric communication opens HD, relieving autoinhibitory (32Dawicki-McKenna DeNizio J.E. Cao C.D. Karch K.R. al.PARP-1 activation local unfolding domain.Mol. 755-768Abstract (204) causes additional WGR-HD interface concomitant concerted rotation (33Rouleau-Turcotte É. Krastev D.B. Pettitt Lord C.J. Captured snapshots state reveal mechanics allostery.Mol. 82: 2939-2951.e5Abstract 2). recognition sequence-dependent allows (SSBs), double-strand (DSBs), even apurinic apyrimidinic integrity backbone preserved 34Khodyreva S.N. Prasad Ilina Sukhanova M.V. Kutuzov M.M. al.Apurinic/apyrimidinic (AP) 5'-dRP/AP lyase polymerase-1 (PARP-1).Proc. Natl. Acad. Sci. U. 107: 22090-22095Crossref contributes chromatin, appear On own, catalytically primarily modifies aspartate glutamate so-called "automodification region" fold nearby linker region (35Ayyappan Wat Barber Vivelo C.A. Gauch Visanpattanasin al.ADPriboDB 2.0: updated database ADP-ribosylated D261-D265Crossref (5) trans other proteins. During DDR, undergoes change specificity collaborates cofactor (HPF1) modify serine histones itself (36Bonfiglio Fontana Zhang Colby Gibbs-Seymour Atanassov al.Serine depends HPF1.Mol. 932-940.e6Abstract (210) newfound ability Ser due joint HPF1, greatly favored HD opening, HPF1 inserts Glu residue deprotonate acceptor initiate (37Suskiewicz Zobel Ogden T.E.H. al.HPF1 completes damage-induced ADP-ribosylation.Nature. 579: 598-602Crossref (139) 38Sun F.H. Zhao Kong L.L. Wong C.C.L. Yun C.H. remodels enable histones.Nat. 1028Crossref (38) being less relies "hit run" form substochiometric ratios (39Langelier Billur Sverzhinsky Black B.E. dynamically controls PARP1/2 initiating elongating modifications.Nat. 6675Crossref (27) Despite short-lived interaction, speeds up initial events reduces sterically blocks Ser-linked appears shorter Glu/Asp-linked modulates shifting Ser-ADP-ribosylation relative automodification 40Gibbs-Seymour HPF1/C4orf27 PARP-1-interacting regulates PARP-1 activity.Mol. 2016; 62: 432-442Abstract (184) ultimately (41Palazzo Leidecker Prokhorova Dauben H. major upon damage.Elife. 7e34334Crossref (63) Overall, burst initiates recruits (i.e., readers). While steered automodifies residues, namely S499, S507, S519 (42Prokhorova Smith Zentout Schutzenhofer al.Serine-linked auto-modification inhibitor response.Nat. 4055Crossref (44) Mutating was shown retain longer suggesting likely needed timely release process. highly negatively charged PTM, charge repulsion driving force (43Murai Huang S.Y. Das B.B. Renaud Doroshow J.H. al.Trapping clinical inhibitors.Cancer 72: 5588-5599Crossref (1497) 44Murai Ji Takeda al.Stereospecific BMN 673 olaparib rucaparib.Mol. Cancer Ther. 2014; 433-443Crossref (565) enacting possible. Another well-studied member closest homolog contrast only short, unstructured N-terminal (NTR) accompany (45Riccio Cingolani PARP-2 requirements localization damage.Nucleic 44: 1691-1702Crossref Also, unlike navigates chromatin. mostly mediated 5′ (46Langelier PARP-3 selective

Language: Английский

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Oncogenic IDH mutations increase heterochromatin-related replication stress without impacting homologous recombination

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(13), P. 2347 - 2356.e8

Published: June 12, 2023

Language: Английский

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Poly(ADP-ribosyl)ation of TIMELESS limits DNA replication stress and promotes stalled fork protection

Cell Reports, Journal Year: 2024, Volume and Issue: 43(3), P. 113845 - 113845

Published: Feb. 22, 2024

Poly(ADP-ribosyl)ation (PARylation), catalyzed mainly by poly(ADP-ribose) polymerase (PARP)1, is a key posttranslational modification involved in DNA replication and repair. Here, we report that TIMELESS (TIM), an essential scaffold of the replisome, PARylated, which linked to its proteolysis. TIM PARylation requires recognition auto-modified PARP1 via two poly(ADP-ribose)-binding motifs, primes for proteasome-dependent degradation. Cells expressing PARylation-refractory mutant or under PARP inhibition accumulate at forks, causing stress hyper-resection stalled forks. Mechanistically, aberrant engagement with replicative helicase impedes RAD51 loading protection reversed Accordingly, defective degradation hypersensitizes BRCA2-deficient cells damage. Our study defines as substrate elucidates how control replisome remodeling fork protection. Therefore, propose mechanism impinges on instability caused turnover.

Language: Английский

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Repriming DNA synthesis: an intrinsic restart pathway that maintains efficient genome replication

Nucleic Acids Research, Journal Year: 2021, Volume and Issue: 49(9), P. 4831 - 4847

Published: March 5, 2021

To bypass a diverse range of fork stalling impediments encountered during genome replication, cells possess variety DNA damage tolerance (DDT) mechanisms including translesion synthesis, template switching, and reversal. These pathways function to obstacles allow efficient synthesis be maintained. In addition, lagging strand can also circumvented by downstream priming Okazaki fragment generation, leaving gaps filled post-replication. Whether repriming occurs on the leading has been intensely debated over past half-century. Early studies indicated that both strands were synthesised discontinuously. Although later suggested was continuous, preferred semi-discontinuous replication model. However, more recently it established replicative primases perform in prokaryotes. An analogous restart mechanism identified most eukaryotes, which specialist primase called PrimPol conducts lesions structures. plays general role maintaining progression. Here, we review discuss historical evidence recent discoveries substantiate as an intrinsic pathway for duplication across all domains life.

Language: Английский

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Mechanisms of PARP1 inhibitor resistance and their implications for cancer treatment

NAR Cancer, Journal Year: 2022, Volume and Issue: 4(4)

Published: Sept. 28, 2022

The discovery of synthetic lethality as a result the combined loss PARP1 and BRCA has revolutionized treatment DNA repair-deficient cancers. With development PARP inhibitors, patients displaying germline or somatic mutations in BRCA1 BRCA2 were presented with novel therapeutic strategy. However, large subset do not respond to inhibitors. Furthermore, many those who eventually acquire resistance. As such, combating de novo acquired resistance inhibitors remains an obstacle achieving durable responses patients. In this review, we touch on some key mechanisms inhibitor resistance, including restoration homologous recombination, replication fork stabilization suppression single-stranded gap accumulation, well address approaches for overcoming

Language: Английский

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TSG101 associates with PARP1 and is essential for PARylation and DNA damage‐induced NF‐κB activation

The EMBO Journal, Journal Year: 2022, Volume and Issue: 41(21)

Published: Sept. 20, 2022

Abstract In a genome‐wide screening for components of the dsDNA‐break‐induced IKK‐NF‐κB pathway, we identified scores regulators, including tumor susceptibility gene TSG101. TSG101 is essential DNA damage‐induced formation cellular poly(ADP‐ribose) (PAR). binds to PARP1 and required activation. This function independent its role in ESCRT‐I endosomal sorting complex. absence TSG101, PAR‐dependent nuclear PARP1‐IKKγ signalosome, which triggers IKK activation, impaired. According requirement NF‐κB TSG101‐deficient cells are defective repair apoptosis protection. Loss results trapping at damage sites mimics effect pharmacological PARP inhibition. We also show that loss connection with inactivated suppressors BRCA1/2 breast cancer lethal. Our imply as therapeutic target achieve synthetic lethality treatment.

Language: Английский

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Human PARP1 substrates and regulators of its catalytic activity: An updated overview

Frontiers in Pharmacology, Journal Year: 2023, Volume and Issue: 14

Published: Feb. 23, 2023

Poly (ADP-ribose) polymerase 1 (PARP1) is a key DNA damage sensor that recruited to damaged sites after strand breaks initiate repair. This achieved by catalyzing attachment of ADP-ribose moieties, which are donated from NAD + , on the amino acid residues itself or other acceptor proteins. PARP inhibitors (PARPi) inhibit catalytic activity and induce trapping commonly used for treating BRCA1/2 -deficient breast ovarian cancers through synergistic lethality. Unfortunately, resistance PARPi frequently occurs. In this review, we present novel substrates regulators PARP1-catalyzed poly (ADP-ribosyl)ation (PARylatison) have been identified in last 3 years. The overall aim presentation protein interactions potential therapeutic intervention overcoming PARPi.

Language: Английский

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ATR protects ongoing and newly assembled DNA replication forks through distinct mechanisms

Cell Reports, Journal Year: 2023, Volume and Issue: 42(7), P. 112792 - 112792

Published: July 1, 2023

The ATR kinase safeguards genomic integrity during S phase, but how protects DNA replication forks remains incompletely understood. Here, we combine four distinct assays to analyze functions at ongoing and newly assembled upon inhibition by hydroxyurea. At forks, inhibitor (ATRi) increases MRE11- EXO1-mediated nascent degradation from PrimPol-generated, single-stranded (ssDNA) gaps. ATRi also exposes template ssDNA through fork uncoupling degradation. Electron microscopy reveals that reduces reversed increasing gap-dependent new triggers CtIP-initiated EXO1, exposing ssDNA. Upon PARP inhibition, preferentially exacerbates in BRCA1/2-deficient cells disrupts the restored gap protection BRCA1-deficient, PARP-inhibitor-resistant cells. Thus, mechanisms, providing an extended view of ATR's stabilizing forks.

Language: Английский

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