Divalent and multivalent cations control liquid-like assembly of poly(ADP-ribosyl)ated PARP1 into multimolecular associates in vitro

• Published in: Communications biology 2024-09, Vol.7 (1), p.1148-17, Article 1148
• Main Authors: Sukhanova, Maria V., Anarbaev, Rashid O., Maltseva, Ekaterina A., Kutuzov, Mikhail M., Lavrik, Olga I.
• Format: Article
• Language: English
• Subjects: 631/45/607/1159; 631/57/2272/2276; 82/16; 82/29; 82/80; 82/83; Biomedical and Life Sciences; Cations - metabolism; Cations, Divalent - metabolism; DNA biosynthesis; DNA Damage; DNA Polymerase beta - metabolism; DNA Repair; DNA-directed DNA polymerase; Glycoside Hydrolases - genetics; Glycoside Hydrolases - metabolism; Humans; Hydrolysis; Life Sciences; Magnesium; Poly (ADP-Ribose) Polymerase-1 - genetics; Poly (ADP-Ribose) Polymerase-1 - metabolism; Poly Adenosine Diphosphate Ribose - metabolism; Poly ADP Ribosylation; Poly(ADP-ribose); Poly(ADP-ribose) glycohydrolase; Poly(ADP-ribose) polymerase; Self-assembly
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-024-06811-4

The formation of nuclear biomolecular condensates is often associated with local accumulation of proteins at a site of DNA damage. The key role in the formation of DNA repair foci belongs to PARP1, which is a sensor of DNA damage and catalyzes the synthesis of poly(ADP-ribose) attracting repair factors. We show here that biogenic cations such as Mg 2+ , Ca 2+ , Mn 2+ , spermidine 3+ , or spermine 4+ can induce liquid-like assembly of poly(ADP-ribosyl)ated [PARylated] PARP1 into multimolecular associates (hereafter: self-assembly). The self-assembly of PARylated PARP1 affects the level of its automodification and hydrolysis of poly(ADP-ribose) by poly(ADP-ribose) glycohydrolase (PARG). Furthermore, association of PARylated PARP1 with repair proteins strongly stimulates strand displacement DNA synthesis by DNA polymerase β (Pol β) but has no noticeable effect on DNA ligase III activity. Thus, liquid-like self-assembly of PARylated PARP1 may play a critical part in the regulation of i) its own activity, ii) PARG-dependent hydrolysis of poly(ADP-ribose), and iii) Pol β–mediated DNA synthesis. The latter can be considered an additional factor influencing the choice between long-patch and short-patch DNA synthesis during repair. Formation of biomolecular condensate via liquid-like assembly of PARylated PARP1 is driven by biogenic cations. This assembly regulates autoPARylation of PARP1, PARG-dependent hydrolysis of poly(ADP-ribose) and Polβ-mediated DNA synthesis.