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

• Published in: Molecular cell Vol. 83; no. 10; pp. 1743 - 1760.e11
• Main Authors: Longarini, Edoardo José, Dauben, Helen, Locatelli, Carolina, Wondisford, Anne R., Smith, Rebecca, Muench, Charlotte, Kolvenbach, Andreas, Lynskey, Michelle Lee, Pope, Alexis, Bonfiglio, Juan José, Jurado, Eva Pinto, Fajka-Boja, Roberta, Colby, Thomas, Schuller, Marion, Ahel, Ivan, Timinszky, Gyula, O’Sullivan, Roderick J., Huet, Sébastien, Matic, Ivan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.05.2023; Cell Press
• Subjects: ADP-Ribosylation; antibodies; Antibodies - genetics; Chromatin; DNA Damage; DNA damage response; Histones - genetics; Histones - metabolism; HPF1; Life Sciences; PARP1; Poly (ADP-Ribose) Polymerase-1 - genetics; Poly (ADP-Ribose) Polymerase-1 - metabolism; RNF114; Signal Transduction; SpyTag; Technology; telomere; HPF1; RNF114; PARP1; DNA damage response; telomere; SpyTag; antibodies; ADP-ribosylation
• ISSN: 1097-2765; 1097-4164; 1097-4164
• DOI: 10.1016/j.molcel.2023.03.027

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 DNA damage, it was unknown whether this signal plays an active role in the cell or is just a byproduct of poly-ADP-ribosylation. By engineering SpyTag-based modular antibodies for sensitive and flexible detection of mono-ADP-ribosylation, including fluorescence-based sensors for live-cell imaging, we demonstrate that serine mono-ADP-ribosylation constitutes a second wave of PARP1 signaling shaped by the cellular HPF1/PARP1 ratio. Multilevel chromatin proteomics reveals histone mono-ADP-ribosylation readers, including RNF114, a ubiquitin ligase recruited to DNA lesions through a zinc-finger domain, modulating the DNA damage response and telomere maintenance. Our work provides a technological framework for illuminating ADP-ribosylation in a wide range of applications and biological contexts and establishes mono-ADP-ribosylation by HPF1/PARP1 as an important information carrier for cell signaling. [Display omitted] •A broadly applicable technology for sensitive and versatile detection of mono-ADPr•Fluorescent probes reveal serine mono-ADPr as a second wave of PARP1 signaling•Multilevel chromatin proteomics identifies histone mono-ADPr readers•RNF114 is a mono-ADPr reader in telomere maintenance and DNA repair signaling Longarini et al. generate high-affinity and sensitivity antibodies for mono-ADP-ribosylation detection in various applications and show that serine mono-ADP-ribosylation constitutes a second wave of PARP1 signaling. They reveal a role of mono-ADPr as a recruitment signal for RNF114, modulating the DNA damage response and telomere maintenance.