Poly(ADP-ribose) binding to Chk1 at stalled replication forks is required for S-phase checkpoint activation
| Autor: | Paulius Grigaravicius, Alexander Bürkle, Fu Li, Anja Krüger, Zhong-Wei Zhou, Zhao-Qi Wang, Chris Meisezahl, Oliver Popp, Bénazir Siddeek, Wookee Min, Christopher Bruhn, Karl-Otto Greulich, Cornelis F. Calkhoven, Xingzhi Xu |
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| Rok vydání: | 2013 |
| Předmět: |
Poly Adenosine Diphosphate Ribose
Amino Acid Motifs genetic processes Poly (ADP-Ribose) Polymerase-1 General Physics and Astronomy medicine.disease_cause environment and public health S Phase Mice PARP1 Chlorocebus aethiops Phosphorylation Polymerase Mutation Multidisciplinary biology Cell Cycle 3T3 Cells Chromatin Recombinant Proteins Cell biology ADP-ribosylation COS Cells Poly(ADP-ribose) Polymerases biological phenomena cell phenomena and immunity Protein Binding DNA Replication animal structures Cell Survival DNA damage Green Fluorescent Proteins Molecular Sequence Data General Biochemistry Genetics and Molecular Biology Cell Line Tumor ddc:570 medicine Animals Humans Amino Acid Sequence Sequence Homology Amino Acid General Chemistry G2-M DNA damage checkpoint Molecular biology enzymes and coenzymes (carbohydrates) HEK293 Cells Checkpoint Kinase 1 biology.protein Protein Kinases DNA Damage |
| Zdroj: | Nature Communications. 4 |
| ISSN: | 2041-1723 |
| Popis: | Damaged replication forks activate poly(ADP-ribose) polymerase 1 (PARP1), which catalyses poly(ADP-ribose) (PAR) formation; however, how PARP1 or poly(ADP-ribosyl)ation is involved in the S-phase checkpoint is unknown. Here we show that PAR, supplied by PARP1, interacts with Chk1 via a novel PAR-binding regulatory (PbR) motif in Chk1, independent of ATR and its activity. iPOND studies reveal that Chk1 associates readily with the unperturbed replication fork and that PAR is required for efficient retention of Chk1 and phosphorylated Chk1 at the fork. A PbR mutation, which disrupts PAR binding, but not the interaction with its partners Claspin or BRCA1, impairs Chk1 and the S-phase checkpoint activation, and mirrors Chk1 knockdown-induced hypersensitivity to fork poisoning. We find that long chains, but not short chains, of PAR stimulate Chk1 kinase activity. Collectively, we disclose a previously unrecognized mechanism of the S-phase checkpoint by PAR metabolism that modulates Chk1 activity at the replication fork. DNA damage at stalled replication forks activates Chk1 kinase and poly(ADP-ribose) (PAR) polymerase 1. Min et al.find that retention of Chk1 to stalled replication forks depends on its direct interaction with PAR, and show that PAR chain length fine-tunes Chk1 and S-phase checkpoint activation. |
| Databáze: | OpenAIRE |
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