Kinetics of poly(ADP-ribosyl)ation, but not PARP1 itself, determines the cell fate in response to DNA damage in vitro and in vivo

H Schuhwerk, C Bruhn, K Siniuk, W Min… - Nucleic acids …, 2017 - academic.oup.com
H Schuhwerk, C Bruhn, K Siniuk, W Min, S Erener, P Grigaravicius, A Krüger, E Ferrari…
Nucleic acids research, 2017academic.oup.com
One of the fastest cellular responses to genotoxic stress is the formation of poly (ADP-ribose)
polymers (PAR) by poly (ADP-ribose) polymerase 1 (PARP1, or ARTD1). PARP1 and its
enzymatic product PAR regulate diverse biological processes, such as DNA repair,
chromatin remodeling, transcription and cell death. However, the inter-dependent function of
the PARP1 protein and its enzymatic activity clouds the mechanism underlying the biological
response. We generated a PARP1 knock-in mouse model carrying a point mutation in the …
Abstract
One of the fastest cellular responses to genotoxic stress is the formation of poly(ADP-ribose) polymers (PAR) by poly(ADP-ribose)polymerase 1 (PARP1, or ARTD1). PARP1 and its enzymatic product PAR regulate diverse biological processes, such as DNA repair, chromatin remodeling, transcription and cell death. However, the inter-dependent function of the PARP1 protein and its enzymatic activity clouds the mechanism underlying the biological response. We generated a PARP1 knock-in mouse model carrying a point mutation in the catalytic domain of PARP1 (D993A), which impairs the kinetics of the PARP1 activity and the PAR chain complexity in vitro and in vivo, designated as hypo-PARylation. PARP1D993A/D993A mice and cells are viable and show no obvious abnormalities. Despite a mild defect in base excision repair (BER), this hypo-PARylation compromises the DNA damage response during DNA replication, leading to cell death or senescence. Strikingly, PARP1D993A/D993A mice are hypersensitive to alkylation in vivo, phenocopying the phenotype of PARP1 knockout mice. Our study thus unravels a novel regulatory mechanism, which could not be revealed by classical loss-of-function studies, on how PAR homeostasis, but not the PARP1 protein, protects cells and organisms from acute DNA damage.
Oxford University Press