Slow Dissociation from the PARP1–HPF1 Complex Drives Inhibitor Potency DOI Creative Commons
Petra Stojanovic, Karolin Luger, Johannes Rudolph

et al.

Biochemistry, Journal Year: 2023, Volume and Issue: 62(16), P. 2382 - 2390

Published: Aug. 2, 2023

PARP1, upon binding to damaged DNA, is activated perform poly ADP-ribosylation (PARylation) on itself and other proteins, which leads relaxation of chromatin recruitment DNA repair factors. HPF1 was recently discovered as a protein cofactor PARP1 that directs preferential PARylation histones over targets by contributing altering the active site. Inhibitors (PARPi) are used in treatment BRCA–/– cancers, but basis for their potency cells, especially context HPF1, not fully understood. Here, we demonstrate simple one-step association eight different PARPi with measured rates (kon) 0.8–6 μM–1 s–1. We find only minor differences these when comparing PARP1–HPF1 complex. By characterizing dissociation (koff) constants (KD) two more PARPi, find, example, saruparib has half-life 22.5 h fluzoparib higher affinity presence just like structurally related compound olaparib. using KD kon calculate koff, cells correlates best koff from Our data suggest drug complex should be parameter choice guiding development next-generation PARPi.

Language: Английский

Inhibitors of PARP: Number crunching and structure gazing DOI Creative Commons
Johannes Rudolph,

Karen Jung,

Karolin Luger

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2022, Volume and Issue: 119(11)

Published: March 8, 2022

SignificancePARP is an important target in the treatment of cancers, particularly patients with breast, ovarian, or prostate cancer that have compromised homologous recombination repair (i.e., BRCA

Language: Английский

Citations

84
ADP-ribosylation from molecular mechanisms to therapeutic implications DOI Creative Commons
Marcin J. Suskiewicz, Evgeniia Prokhorova, J.G.M. Rack

et al.

Cell, Journal Year: 2023, Volume and Issue: 186(21), P. 4475 - 4495

Published: Oct. 1, 2023

ADP-ribosylation is a ubiquitous modification of biomolecules, including proteins and nucleic acids, that regulates various cellular functions in all kingdoms life. The recent emergence new technologies to study has reshaped our understanding the molecular mechanisms govern establishment, removal, recognition this modification, as well its impact on organismal function. These advances have also revealed intricate involvement human physiology pathology enormous potential their manipulation holds for therapy. In review, we present state-of-the-art findings covering work structural biology, biochemistry, cell clinical aspects ADP-ribosylation.

Language: Английский

Citations

70
HPF1-dependent histone ADP-ribosylation triggers chromatin relaxation to promote the recruitment of repair factors at sites of DNA damage DOI
Rebecca Smith, Siham Zentout,

Magdalena B. Rother

et al.

Nature Structural & Molecular Biology, Journal Year: 2023, Volume and Issue: 30(5), P. 678 - 691

Published: April 27, 2023

Language: Английский

Citations

45
Poly(ADP-ribosyl)ation enhances nucleosome dynamics and organizes DNA damage repair components within biomolecular condensates DOI Creative Commons
Michael L. Nosella, Tae Hun Kim, Shuya Kate Huang

et al.

Molecular Cell, Journal Year: 2024, Volume and Issue: 84(3), P. 429 - 446.e17

Published: Jan. 11, 2024

Language: Английский

Citations

19
The regulatory landscape of the human HPF1- and ARH3-dependent ADP-ribosylome DOI Creative Commons
Ivo A. Hendriks, Sara C. Buch-Larsen, Evgeniia Prokhorova

et al.

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: Oct. 8, 2021

Abstract Despite the involvement of Poly(ADP-ribose) polymerase-1 (PARP1) in many important biological pathways, target residues PARP1-mediated ADP-ribosylation remain ambiguous. To explicate regulome, we analyze human cells depleted for key regulators PARP1 activity, histone PARylation factor 1 (HPF1) and ADP-ribosylhydrolase 3 (ARH3). Using quantitative proteomics, characterize 1,596 sites, displaying up to 1000-fold regulation across investigated knockout cells. We find that HPF1 ARH3 inversely homogenously regulate serine ADP-ribosylome on a proteome-wide scale with consistent adherence lysine-serine-motifs, suggesting targeting is independent ARH3. Notably, do not detect an HPF1-dependent residue switch from glutamate/aspartate under conditions. Our data support notion mainly exists as mono-ADP-ribosylation cells, reveal remarkable degree co-modification other post-translational modifications.

Language: Английский

Citations

80
Serine-linked PARP1 auto-modification controls PARP inhibitor response DOI Creative Commons
Evgeniia Prokhorova,

Florian Zobel,

Rebecca Smith

et al.

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: July 1, 2021

Abstract Poly(ADP-ribose) polymerase 1 (PARP1) and PARP2 are recruited activated by DNA damage, resulting in ADP-ribosylation at numerous sites, both within PARP1 itself other proteins. Several inhibitors currently employed the clinic or undergoing trials for treatment of various cancers. These drugs act primarily trapping on damaged chromatin, which can lead to cell death, especially cells with repair defects. Although is thought be caused catalytic inhibition PARP-dependent modification, implying that (ADPr) counteract trapping, it not known exact sites important this process. Following recent findings PARP1- PARP2-mediated modification predominantly serine-linked, we demonstrate here serine ADPr plays a vital role cellular responses PARP1/PARP2 inhibitors. Specifically, identify three residues (499, 507, 519) as key whose efficient HPF1-dependent counters contributes inhibitor tolerance. Our data implicate genes encode serine-specific regulators, HPF1 ARH3, potential therapy biomarkers.

Language: Английский

Citations

76
The BRCT domain of PARP1 binds intact DNA and mediates intrastrand transfer DOI Creative Commons
Johannes Rudolph, Uma M. Muthurajan, Megan Palacio

et al.

Molecular Cell, Journal Year: 2021, Volume and Issue: 81(24), P. 4994 - 5006.e5

Published: Dec. 1, 2021

Language: Английский

Citations

72
PARP inhibitors trap PARP2 and alter the mode of recruitment of PARP2 at DNA damage sites DOI Creative Commons
Xiaohui Lin, Wenxia Jiang, Johannes Rudolph

et al.

Nucleic Acids Research, Journal Year: 2022, Volume and Issue: 50(7), P. 3958 - 3973

Published: March 23, 2022

Dual-inhibitors of PARP1 and PARP2 are promising anti-cancer drugs. In addition to blocking PARP1&2 enzymatic activity, PARP inhibitors also extend the lifetime DNA damage-induced foci, termed trapping. Trapping is important for therapeutic effects inhibitors. Using live-cell imaging, we found that cause persistent foci by switching mode recruitment from a predominantly PARP1- PAR-dependent rapid exchange WGR domain-mediated stalling on DNA. Specifically, PARP1-deletion markedly reduces but does not abolish foci. The residual in PARP1-deficient cells DNA-dependent abrogated R140A mutation domain. Yet, PARP2-R140A forms normal PARP1-proficient cells. cells, - niraparib, talazoparib, and, lesser extent, olaparib enhance preventing exchange. This trapping independent auto-PARylation abolished domain H415A catalytic Taken together, trap physically via WGR-DNA interaction while suppressing PARP2.

Language: Английский

Citations

41
PARPs and ADP-ribosylation: Deciphering the complexity with molecular tools DOI Creative Commons
Morgan Dasovich, Anthony K. L. Leung

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(10), P. 1552 - 1572

Published: April 28, 2023

Language: Английский

Citations

34
The logic of protein post‐translational modifications (PTMs): Chemistry, mechanisms and evolution of protein regulation through covalent attachments DOI Creative Commons
Marcin J. Suskiewicz

BioEssays, Journal Year: 2024, Volume and Issue: 46(3)

Published: Jan. 21, 2024

Abstract Protein post‐translational modifications (PTMs) play a crucial role in all cellular functions by regulating protein activity, interactions and half‐life. Despite the enormous diversity of modifications, various PTM systems show parallels their chemical catalytic underpinnings. Here, focussing on that involve addition new elements to amino‐acid sidechains, I describe historical milestones fundamental concepts support current understanding PTMs. The survey covers selected key research programmes, including study phosphorylation as regulatory switch, ubiquitylation degradation signal histone functional code. contribution techniques for studying PTMs is also discussed. central part essay explores shared principles strategies observed across diverse systems, together with mechanisms substrate selection, reversibility erasers recognition reader domains. Similarities basic mechanism are highlighted implications final dedicated evolutionary trajectories beginning possible emergence context rivalry prokaryotic world. Together, provides unified perspective world major modifications.

Language: Английский

Citations

9