The expanding universe of PARP1-mediated molecular and therapeutic mechanisms

Molecular Cell, Год журнала: 2022, Номер 82(12), С. 2315 - 2334

Опубликована: Март 9, 2022

Язык: Английский

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PARP1-DNA co-condensation drives DNA repair site assembly to prevent disjunction of broken DNA ends

Cell, Год журнала: 2024, Номер 187(4), С. 945 - 961.e18

Опубликована: Фев. 1, 2024

DNA double-strand breaks (DSBs) are repaired at DSB sites. How sites assemble and how broken is prevented from separating not understood. Here we uncover that the synapsis of mediated by sensor protein poly(ADP-ribose) (PAR) polymerase 1 (PARP1). Using bottom-up biochemistry, reconstitute functional show form through co-condensation PARP1 multimers with DNA. The co-condensates exert mechanical forces to keep ends together become enzymatically active for PAR synthesis. PARylation promotes release recruitment effectors, such as Fused in Sarcoma, which stabilizes against separation, revealing a finely orchestrated order events primes repair. We provide comprehensive model hierarchical assembly condensates explain end effector proteins damage

Язык: Английский

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HPF1-dependent histone ADP-ribosylation triggers chromatin relaxation to promote the recruitment of repair factors at sites of DNA damage

Nature Structural & Molecular Biology, Год журнала: 2023, Номер 30(5), С. 678 - 691

Опубликована: Апрель 27, 2023

Язык: Английский

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Immediate-Early, Early, and Late Responses to DNA Double Stranded Breaks

Frontiers in Genetics, Год журнала: 2022, Номер 13

Опубликована: Янв. 31, 2022

Loss or rearrangement of genetic information can result from incorrect responses to DNA double strand breaks (DSBs). The cellular DSBs encompass a range highly coordinated events designed detect and respond appropriately the damage, thereby preserving genomic integrity. In analogy with occurring during viral infection, we appropriate terms Immediate-Early, Early, Late describe pre-repair DSBs. A distinguishing feature Immediate-Early response is that large protein condensates form Early are resolved upon repair, termed foci, not visible. encompasses initial lesion sensing, involving poly (ADP-ribose) polymerases (PARPs), KU70/80, MRN, as well rapid repair by so-called 'fast-kinetic' canonical non-homologous end joining (cNHEJ). Initial binding PARPs KU70/80 complex appears be mutually exclusive at easily ligatable repaired efficiently fast-kinetic cNHEJ; process PARP-, ATM-, 53BP1-, Artemis-, resection-independent. However, more requiring processing, ensuing dynamic PARylation (polyADP ribosylation) many substrates may aid recruitment both MRN Complex rely response, largely defined ATM-dependent focal signalling molecules into condensates, regulated chromatin dynamics. Finally, integrates cell cycle phase, context, type DSB determine pathway choice. Critical choice p53 1 (53BP1) breast cancer associated (BRCA1). additional factors recruited throughout also impact choice, although these remain fully characterised. somehow channels high-fidelity pathway, typically either 'slow-kinetic' cNHEJ homologous recombination (HR). specific components machinery results in cells utilising remaining effect but often cost increased mutagenesis. Here discuss regulation proceeding itself.

Язык: Английский

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Captured snapshots of PARP1 in the active state reveal the mechanics of PARP1 allostery

Molecular Cell, Год журнала: 2022, Номер 82(16), С. 2939 - 2951.e5

Опубликована: Июль 5, 2022

Язык: Английский

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A two-step mechanism governing PARP1-DNA retention by PARP inhibitors

Science Advances, Год журнала: 2022, Номер 8(36)

Опубликована: Сен. 7, 2022

PARP inhibitors (PARPi) have emerged as promising cancer therapeutics capable of targeting specific DNA repair pathways, but their mechanism action with respect to PARP1-DNA retention remains unclear. Here, we developed single-molecule assays directly monitor the PARP1 on lesions in real time. Our study reveals a two-step by which PARPi modulate lesions, consisting primary step catalytic inhibition via binding competition NAD + followed an allosteric modulation bound PARPi. While clinically relevant exhibit distinct activities that can either increase or induce its release, potencies are predominantly determined ability outcompete binding. These findings provide mechanistic basis for improved selection according characteristic and enable further development more potent inhibitors.

Язык: Английский

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Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling

Molecular Cell, Год журнала: 2023, Номер 83(10), С. 1743 - 1760.e11

Опубликована: Апрель 27, 2023

PARP1, an established anti-cancer target that regulates many cellular pathways, including DNA repair signaling, has been intensely studied for decades as a poly(ADP-ribosyl)transferase. Although recent studies have revealed the prevalence of mono-ADP-ribosylation upon damage, it was unknown whether this signal plays active role in cell or is just byproduct poly-ADP-ribosylation. By engineering SpyTag-based modular antibodies sensitive and flexible detection mono-ADP-ribosylation, fluorescence-based sensors live-cell imaging, we demonstrate serine constitutes second wave PARP1 signaling shaped by HPF1/PARP1 ratio. Multilevel chromatin proteomics reveals histone readers, RNF114, ubiquitin ligase recruited to lesions through zinc-finger domain, modulating damage response telomere maintenance. Our work provides technological framework illuminating ADP-ribosylation wide range applications biological contexts establishes important information carrier signaling.

Язык: Английский

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PARPs and ADP-ribosylation: Deciphering the complexity with molecular tools

Molecular Cell, Год журнала: 2023, Номер 83(10), С. 1552 - 1572

Опубликована: Апрель 28, 2023

Язык: Английский

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Clinical PARP inhibitors allosterically induce PARP2 retention on DNA

Science Advances, Год журнала: 2023, Номер 9(12)

Опубликована: Март 24, 2023

PARP1 and PARP2 detect DNA breaks, which activates their catalytic production of poly(ADP-ribose) that recruits repair factors contributes to PARP1/2 release from DNA. PARP inhibitors (PARPi) are used in cancer treatment target activity, interfering with increasing persistence on damage. In addition, certain PARPi exert allosteric effects increase retention However, no clinical exhibit this behavior toward PARP1. contrast, we show an effect retains breaks a manner depends communication between the binding regions. Using mutant mimics inhibitor effect, observed increased at cellular damage sites. The AZD5305 also exhibited clear reverse PARP2. Our results can help explain toxicity suggest ways improve moving forward.

Язык: Английский

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Chemoenzymatic and Synthetic Approaches To Investigate Aspartate- and Glutamate-ADP-Ribosylation

Journal of the American Chemical Society, Год журнала: 2023, Номер 145(25), С. 14000 - 14009

Опубликована: Июнь 14, 2023

We report here chemoenzymatic and fully synthetic methodologies to modify aspartate glutamate side chains with ADP-ribose at specific sites on peptides. Structural analysis of ADP-ribosylated peptides reveals near-quantitative migration the chain linkage from anomeric carbon 2″- or 3″-ADP-ribose hydroxyl moieties. find that this pattern is unique ADP-ribosylation propose observed isomer distribution profile present in biochemical cellular environments. After defining distinct stability properties ADP-ribosylation, we devise methods install homogenous assemble glutamate-modified into full-length proteins. By implementing these technologies, show histone H2B E2 tri-ADP-ribosylation able stimulate chromatin remodeler ALC1 similar efficiency serine ADP-ribosylation. Our work fundamental principles enables new strategies interrogate consequences widespread protein modification.

Язык: Английский

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