Repairing split ends: SIRT6, mono-ADP ribosylation and DNA repair

AbstractUpon binding to DNA breaks, poly(ADP-ribose) polymerase 1 (PARP1) ADP-ribosylates itself and other factors to initiate DNA repair. Serine is the major residue for ADP-ribosylation upon DNA damage, which strictly depends on HPF1. Here, we report the crystal structures of human HPF1/PARP1-CAT ΔHD complex at 1.98 Å resolution, and mouse and human HPF1 at 1.71 Å and 1.57 Å resolution, respectively. Our structures and mutagenesis data confirm that the structural insights obtained in a recent HPF1/PARP2 study by Suskiewicz et al. apply to PARP1. Moreover, we quantitatively characterize the key residues necessary for HPF1/PARP1 binding. Our data show that through salt-bridging to Glu284/Asp286, Arg239 positions Glu284 to catalyze serine ADP-ribosylation, maintains the local conformation of HPF1 to limit PARP1 automodification, and facilitates HPF1/PARP1 binding by neutralizing the negative charge of Glu284. These findings, along with the high-resolution structural data, may facilitate drug discovery targeting PARP1.

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RAP80 and BRCA1 PARsylation protect chromosome integrity by preventing retention of BRCA1-B/C complexes in DNA repair foci

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1908003117 ◽

2020 ◽

Vol 117 (4) ◽

pp. 2084-2091

Author(s):

Jekaterina Vohhodina ◽

Kimberly J. Toomire ◽

Sarah A. Petit ◽

Goran Micevic ◽

Geeta Kumari ◽

...

Keyword(s):

Dna Repair ◽

Homologous Recombination ◽

Complex Formation ◽

Illegitimate Recombination ◽

Adp Ribosylation ◽

Chromosomal Disorder ◽

Nuclear Foci ◽

Simultaneous Loss ◽

Chromosome Integrity ◽

Focal Structures

BRCA1 promotes error-free, homologous recombination-mediated repair (HRR) of DNA double-stranded breaks (DSBs). When excessive and uncontrolled, BRCA1 HRR activity promotes illegitimate recombination and genome disorder. We and others have observed that the BRCA1-associated protein RAP80 recruits BRCA1 to postdamage nuclear foci, and these chromatin structures then restrict the amplitude of BRCA1-driven HRR. What remains unclear is how this process is regulated. Here we report that both BRCA1 poly-ADP ribosylation (PARsylation) and the presence of BRCA1-bound RAP80 are critical for the normal interaction of BRCA1 with some of its partners (e.g., CtIP and BACH1) that are also known components of the aforementioned focal structures. Surprisingly, the simultaneous loss of RAP80 and failure therein of BRCA1 PARsylation results in the dysregulated accumulation in these foci of BRCA1 complexes. This in turn is associated with the intracellular development of a state of hyper-recombination and gross chromosomal disorder. Thus, physiological RAP80-BRCA1 complex formation and BRCA1 PARsylation contribute to the kinetics by which BRCA1 HRR-sustaining complexes normally concentrate in nuclear foci. These events likely contribute to aneuploidy suppression.

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ADP-Ribosylation, DNA Repair, and Chromatin Organization

Proceedings in Life Sciences - ADP-Ribosylation of Proteins ◽

10.1007/978-3-642-70589-2_31 ◽

1985 ◽

pp. 235-241 ◽

Cited By ~ 4

Author(s):

Felix R. Althaus ◽

Georg Mathis

Keyword(s):

Dna Repair ◽

Chromatin Organization ◽

Adp Ribosylation

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ADP-RIBOSYLATION AND DNA REPAIR AS INDUCED BY NITROSOUREAS

Nitrosoureas ◽

10.1016/b978-0-12-565060-1.50014-6 ◽

1981 ◽

pp. 123-142

Author(s):

Mark Smulson ◽

Tauseef Butt ◽

Swaroop Sudhakar ◽

Don Jump ◽

Nancy Nolan

Keyword(s):

Dna Repair ◽

Adp Ribosylation

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PARP1-Driven Poly-ADP-Ribosylation Regulates BRCA1 Function in Homologous Recombination–Mediated DNA Repair

Cancer Discovery ◽

10.1158/2159-8290.cd-13-0891 ◽

2014 ◽

Vol 4 (12) ◽

pp. 1430-1447 ◽

Cited By ~ 62

Author(s):

Yiduo Hu ◽

Sarah A. Petit ◽

Scott B. Ficarro ◽

Kimberly J. Toomire ◽

Anyong Xie ◽

...

Keyword(s):

Dna Repair ◽

Homologous Recombination ◽

Adp Ribosylation

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Reversible ADP-ribosylation of RNA

Nucleic Acids Research ◽

10.1093/nar/gkz305 ◽

2019 ◽

Vol 47 (11) ◽

pp. 5658-5669 ◽

Cited By ~ 33

Author(s):

Deeksha Munnur ◽

Edward Bartlett ◽

Petra Mikolčević ◽

Ilsa T Kirby ◽

Johannes Gregor Matthias Rack ◽

...

Keyword(s):

Dna Repair ◽

Stress Response ◽

Nicotinamide Adenine Dinucleotide ◽

Cellular Stress Response ◽

Biological Processes ◽

Adp Ribosylation ◽

Human Proteins ◽

Novel Target ◽

Host Virus Interactions ◽

Virus Interactions

Abstract ADP-ribosylation is a reversible chemical modification catalysed by ADP-ribosyltransferases such as PARPs that utilize nicotinamide adenine dinucleotide (NAD+) as a cofactor to transfer monomer or polymers of ADP-ribose nucleotide onto macromolecular targets such as proteins and DNA. ADP-ribosylation plays an important role in several biological processes such as DNA repair, transcription, chromatin remodelling, host-virus interactions, cellular stress response and many more. Using biochemical methods we identify RNA as a novel target of reversible mono-ADP-ribosylation. We demonstrate that the human PARPs - PARP10, PARP11 and PARP15 as well as a highly diverged PARP homologue TRPT1, ADP-ribosylate phosphorylated ends of RNA. We further reveal that ADP-ribosylation of RNA mediated by PARP10 and TRPT1 can be efficiently reversed by several cellular ADP-ribosylhydrolases (PARG, TARG1, MACROD1, MACROD2 and ARH3), as well as by MACROD-like hydrolases from VEEV and SARS viruses. Finally, we show that TRPT1 and MACROD homologues in bacteria possess activities equivalent to the human proteins. Our data suggest that RNA ADP-ribosylation may represent a widespread and physiologically relevant form of reversible ADP-ribosylation signalling.

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Serine ADP-ribosylation reversal by the hydrolase ARH3

eLife ◽

10.7554/elife.28533 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 72

Author(s):

Pietro Fontana ◽

Juan José Bonfiglio ◽

Luca Palazzo ◽

Edward Bartlett ◽

Ivan Matic ◽

...

Keyword(s):

Dna Damage ◽

Dna Repair ◽

Posttranslational Modification ◽

Quantitative Proteomics ◽

Chromatin Regulation ◽

Adp Ribosylation ◽

Cellular Processes ◽

New Form ◽

Hydrolysis Of ◽

In Cells

ADP-ribosylation (ADPr) is a posttranslational modification (PTM) of proteins that controls many cellular processes, including DNA repair, transcription, chromatin regulation and mitosis. A number of proteins catalyse the transfer and hydrolysis of ADPr, and also specify how and when the modification is conjugated to the targets. We recently discovered a new form of ADPr that is attached to serine residues in target proteins (Ser-ADPr) and showed that this PTM is specifically made by PARP1/HPF1 and PARP2/HPF1 complexes. In this work, we found by quantitative proteomics that histone Ser-ADPr is reversible in cells during response to DNA damage. By screening for the hydrolase that is responsible for the reversal of Ser-ADPr, we identified ARH3/ADPRHL2 as capable of efficiently and specifically removing Ser-ADPr of histones and other proteins. We further showed that Ser-ADPr is a major PTM in cells after DNA damage and that this signalling is dependent on ARH3.

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Serine-linked PARP1 auto-modification controls PARP inhibitor response

Nature Communications ◽

10.1038/s41467-021-24361-9 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Evgeniia Prokhorova ◽

Florian Zobel ◽

Rebecca Smith ◽

Siham Zentout ◽

Ian Gibbs-Seymour ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Dna Repair ◽

Parp Inhibitor ◽

Cellular Responses ◽

Vital Role ◽

Inhibitor Tolerance ◽

Adp Ribosylation ◽

Key Sites ◽

Dependent Modification

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

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