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Overlapping roles for PARP1 and PARP2 in the recruitment of endogenous XRCC1 and PNKP into oxidized chromatin

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posted on 2023-06-09, 07:43 authored by Hana Hanzlikova, William GittensWilliam Gittens, Katerina Krejcikova, Zhihong Zeng, Keith CaldecottKeith Caldecott
A critical step of DNA single-strand break repair is the rapid recruitment of the scaffold protein XRCC1 that interacts with, stabilizes and stimulates multiple enzymatic components of the repair process. XRCC1 recruitment is promoted by PARP1, an enzyme that is activated following DNA damage and synthesizes ADP-ribose polymers that XRCC1 binds directly. However, cells possess two other DNA strand breakinduced PARP enzymes, PARP2 and PARP3, for which the roles are unclear. To address their involvement in the recruitment of endogenous XRCC1 into oxidized chromatin we have established ‘isogenic’ human diploid cells in which PARP1 and/or PARP2, or PARP3 are deleted. Surprisingly, we show that either PARP1 or PARP2 are sufficient for near-normal XRCC1 recruitment at oxidative single-strand breaks (SSBs) as indicated by the requirement for loss of both proteins to greatly reduce or ablate XRCC1 chromatin binding following H2O2 treatment. Similar results were observed for PNKP; an XRCC1 protein partner important for repair of oxidative SSBs. Notably,concentrations of PARP inhibitor >1000-fold higher than the IC50 were required to ablate both ADP-ribosylation and XRCC1 chromatin binding following H2O2 treatment. These results demonstrate that very low levels of ADP-ribosylation, synthesized by either PARP1 or PARP2, are sufficient for XRCC1 recruitment following oxidative stress.

Funding

Cellular and Pathological Responses to Chromosome DNA Single-Strand Breaks; G2053; MRC-MEDICAL RESEARCH COUNCIL; MR/P010121/1

Chromosomal Single-strand break Repair: Mechanisms & Degenerative Disease; G0830; MRC-MEDICAL RESEARCH COUNCIL; MR/J006750/1

History

Publication status

  • Published

File Version

  • Published version

Journal

Nucleic Acids Research

ISSN

1362-4962

Publisher

Oxford Journals

Issue

5

Volume

45

Page range

2546-2557

Department affiliated with

  • Sussex Centre for Genome Damage Stability Publications

Research groups affiliated with

  • Genome Damage and Stability Centre Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2017-08-30

First Open Access (FOA) Date

2017-08-30

First Compliant Deposit (FCD) Date

2017-08-30

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