Involvement of Poly(ADP-ribose) Polymerase-1 and XRCC1/DNA Ligase III in an Alternative Route for DNA Double-strand Breaks Rejoining*

By M. Audebert, B. Salles, P. Calsou. Journal of Biological Chemistry, 2004

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The efficient repair of DNA double-strand breaks (DSBs) is critical for the maintenance of genomic integrity. In mammalian cells, the nonhomologous end-joining process that represents the predominant repair pathway relies on the DNA-dependent protein kinase (DNA-PK) and the XRCC4-DNA ligase IV complex. Nonetheless, several in vitro and in vivo results indicate that mammalian cells use more than a single end-joining mechanism. While searching for a DNA-PK-independent end-joining activity, we found that the pretreatment of DNA-PK-proficient and -deficient rodent cells with an inhibitor of the poly(ADP-ribose) polymerase-1 enzyme (PARP-1) led to increased cytotoxicity of the highly efficient DNA double-strand breaking compound calicheamicin γ1. In addition, the repair kinetics of the DSBs induced by calicheamicin γ1 was delayed both in PARP-1-proficient cells pretreated with the PARP-1 inhibitor and in PARP-1-deficient cells. In order to get new insights into the mechanism of an alternative route for DSBs repair, we have established a new synapsis and end-joining two-step assay in vitro, operating on DSBs with either nuclear protein extracts or recombinant proteins. We found an end-joining activity independent of the DNA-PK/XRCC4-ligase IV complex but that actually required a novel synapsis activity of PARP-1 and the ligation activity of the XRCC1-DNA ligase III complex, proteins otherwise involved in the base excision repair pathway. Taken together, these results strongly suggest that a PARP-1-dependent DSBs end-joining activity may exist in mammalian cells. We propose that this mechanism could act as an alternative route of DSBs repair that complements the DNA-PK/XRCC4/ligase IV-dependent nonhomologous end-joining.

Subfields: Molecular Biology, Oncology

View Paper: https://www.semanticscholar.org/paper/88933f90f4532185165e32b3b5a9458c1c91a506

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Citations in the corpus

Chapter 1. General introduction (2015, unknown): “# e synapsis activity of Parp1 and the ligation activity of Lig3/ XRCC1 were established via a two-step DNA in vitro pull down assay with nuclear extracts and recombinant proteins (85).”
PARP inhibition: PARP1 and beyond (2010, background): “Additional in vitro studies in rodent and human cells have implicated PARP1 in non-homologous end joining (NHEJ”
PARP-1 and Ku compete for repair of DNA double strand breaks by distinct NHEJ pathways (2006, background): “This suggests that rejoining of DSBs remains possible in the absence of PARP-1 and may be carried out by DNA ligase III, possibly aided by its zinc finger domain (21,22).”
The histone methyltransferase SMYD2 methylates PARP1 and promotes poly(ADP-ribosyl)ation activity in cancer cells. (2014, background): “PARP1 has been shown to play multiple critical roles in the repair of DNA single-strand breaks (SSBs) and double strand breaks (DSBs) [4-9].”
PARPs and ADP-ribosylation: recent advances linking molecular functions to biological outcomes (2017, background): “PARP-1 and DNA repair PARP-1 is recruited to nicks and DSBs in genomic DNA in response to DNA damage and is a criticalmediator of DNA damage repair (Audebert et al. 2004; Langelier et al. 2012).”
MMEJ repair of double-strand breaks (director’s cut): deleted sequences and alternative endings (2008, background): “PARP-1 Human and rodent cell lines Synapsis of ends BER [ 52 ,54] Histone H1 Human cell lines Unclear, stimulated or inhibited MMEJ in”
Homologous recombination deficiency: Exploiting the fundamental vulnerability of ovarian cancer (2015, background): “Molecular characterization of alt-EJ revealed that the XRCC1/ DNA ligase III complex and PARP1 were involved (84, 117, 118).”
PARP-1 cleavage fragments: signatures of cell-death proteases in neurodegeneration (2010, background): “Initially identified as an enzyme that performs central roles in the repair of damaged DNA, PARP-1 participates in initiating base excision repair (BER) (PARP-1-/- cells have impaired BER activity) [1,2], nucleotide excision repair, single strand base repair mediated by DNA ligase III, XRCC1, poly n”