XRCC1 protects transcription from toxic PARP1 activity during DNA base excision repair

Adamowicz, Marek; Hailstone, Richard; Demin, Annie Albert; Komulainen, Emilia; Hanzlíková, Hana; Bražina, Jan; Gautam, Amit; Wells, Sophie E; Caldecott, Keith W.

doi:10.1038/s41556-021-00792-w

Cited by 38 publications

(20 citation statements)

References 92 publications

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“…Data from the present study combined with the one by the Caldecott lab (Adamowicz et al 2021;Demin et al 2021) are consistent with and extend a model that has previously been proposed by Dianov and Hübscher (Dianov and Hubscher 2013). According to this model, one of the key functions of PARP1 during SSBR comprises the regulation of SSBR capacity and prevention of DSB formation by nuclease attack.…”

Section: Discussionsupporting

confidence: 92%

PARP1 and XRCC1 exhibit a reciprocal relationship in genotoxic stress response

et al. 2022

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PARP1 (aka ARTD1) acts as a prime sensor of cellular genotoxic stress response. PARP1 detects DNA strand breaks and subsequently catalyzes the formation of poly(ADP-ribose) (PAR), which leads to the recruitment of the scaffold protein XRCC1 during base excision and single strand break repair and the assembly of multi-protein complexes to promote DNA repair. Here, we reveal that the recruitment of either protein to sites of DNA damage is impeded in the absence of the other, indicating a strong reciprocal relationship between the two DNA repair factors during genotoxic stress response. We further analyzed several cellular and molecular endpoints in HeLa PARP1 KO, XRCC1 KO, and PARP1/XRCC1 double KO (DKO) cells after genotoxic treatments, i.e., PARylation response, NAD+ levels, clonogenic survival, cell cycle progression, cell death, and DNA repair. The analysis of NAD+ levels and cytotoxicity after treatment with the topoisomerase I inhibitor camptothecin revealed a hypersensitivity phenotype of XRCC1 KO cells compared to PARP1 KO cells—an effect that could be rescued by the additional genetic deletion of PARP1 as well as by pharmacological PARP inhibition. Moreover, impaired repair of hydrogen peroxide and CPT-induced DNA damage in XRCC1 KO cells could be partially rescued by additional deletion of PARP1. Our results therefore highlight important reciprocal regulatory functions of XRCC1 and PARP1 during genotoxic stress response.

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Section: Discussionsupporting

confidence: 92%

PARP1 and XRCC1 exhibit a reciprocal relationship in genotoxic stress response

et al. 2022

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“…We showed that the loss of XRCC1 causes PARP1 to exert its intrinsic poisoning activity, leading to the rapid NAD + depletion during treatment with TMZ (Figure 5b). NAD + depletion is seen in various diseases, including the postischemic injury of the heart, brain, liver, and kidney, neurodegeneration, and cancer (Adamowicz et al, 2021; Gujar et al, 2016; Hoch et al, 2017; Pieper et al, 2000). NAD + depletion by its biosynthesis blockade is used to treat cancers and interferes with BER of TMZ‐induced base damage (Goellner et al, 2011; Touat et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

XRCC1 counteracts poly(ADP ribose)polymerase (PARP) poisons, olaparib and talazoparib, and a clinical alkylating agent, temozolomide, by promoting the removal of trapped PARP1 from broken DNA

et al. 2022

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Base excision repair (BER) removes damaged bases by generating single-strand breaks (SSBs), gap-filling by DNA polymerase β (POLβ), and resealing SSBs. A base-damaging agent, methyl methanesulfonate (MMS) is widely used to study BER. BER increases cellular tolerance to MMS, anti-cancer base-damaging drugs, temozolomide, carmustine, and lomustine, and to clinical poly(ADP ribose)polymerase (PARP) poisons, olaparib and talazoparib. The poisons stabilize PARP1/SSB complexes, inhibiting access of BER factors to SSBs. PARP1 and XRCC1 collaboratively promote SSB resealing by recruiting POLβ to SSBs, but XRCC1 À/À cells are much more sensitive to MMS than PARP1 À/À cells. We recently report that the PARP1 loss in XRCC1 À/À cells restores their MMS tolerance and conclude that XPCC1 facilitates the release of PARP1 from SSBs by maintaining its autoPARylation. We here show that the PARP1 loss in XRCC1 À/À cells also restores their tolerance to the three anti-cancer basedamaging drugs, although they and MMS induce different sets of base damage.We reveal the synthetic lethality of the XRCC1 À/À mutation, but not POLβ À/À , with olaparib and talazoparib, indicating that XRCC1 is a unique BER factor in suppressing toxic PARP1/SSB complex and can suppress even when PARP1

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“…While the rapid localisation of PARP1 to single-strand breaks has been well characterised, its precise mechanism of localisation to DSBs remains unknown (Liu et al, 2017). In SSB repair, PARP1 and its associated PARylation events recruit X-ray repair crosscomplementing protein 1 (XRCC1) to SSB sites, with XRCC1 functioning as a scaffold for the subsequent binding of SSB repair proteins (Masson et al, 1998;Breslin et al, 2015;Hanzlikova et al, 2017;Adamowicz et al, 2021). It is currently unclear how the cell might differentiate between isolated SSBs and those that occur in very close proximity but on opposite strand (i.e., DSBs) after the initial PARylation.…”

Section: The Role Of Parp In the Immediate-early Double Strand Break Responsementioning

confidence: 99%

Immediate-Early, Early, and Late Responses to DNA Double Stranded Breaks

Kieffer

Lowndes

2022

Front. Genet.

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Loss or rearrangement of genetic information can result from incorrect responses to DNA double strand breaks (DSBs). The cellular responses to DSBs encompass a range of highly coordinated events designed to detect and respond appropriately to the damage, thereby preserving genomic integrity. In analogy with events occurring during viral infection, we appropriate the terms Immediate-Early, Early, and Late to describe the pre-repair responses to DSBs. A distinguishing feature of the Immediate-Early response is that the large protein condensates that form during the Early and Late response and are resolved upon repair, termed foci, are not visible. The Immediate-Early response encompasses initial lesion sensing, involving poly (ADP-ribose) polymerases (PARPs), KU70/80, and MRN, as well as rapid repair by so-called ‘fast-kinetic’ canonical non-homologous end joining (cNHEJ). Initial binding of PARPs and the KU70/80 complex to breaks appears to be mutually exclusive at easily ligatable DSBs that are repaired efficiently by fast-kinetic cNHEJ; a process that is PARP-, ATM-, 53BP1-, Artemis-, and resection-independent. However, at more complex breaks requiring processing, the Immediate-Early response involving PARPs and the ensuing highly dynamic PARylation (polyADP ribosylation) of many substrates may aid recruitment of both KU70/80 and MRN to DSBs. Complex DSBs rely upon the Early response, largely defined by ATM-dependent focal recruitment of many signalling molecules into large condensates, and regulated by complex chromatin dynamics. Finally, the Late response integrates information from cell cycle phase, chromatin context, and type of DSB to determine appropriate pathway choice. Critical to pathway choice is the recruitment of p53 binding protein 1 (53BP1) and breast cancer associated 1 (BRCA1). However, additional factors recruited throughout the DSB response also impact upon pathway choice, although these remain to be fully characterised. The Late response somehow channels DSBs into the appropriate high-fidelity repair pathway, typically either ‘slow-kinetic’ cNHEJ or homologous recombination (HR). Loss of specific components of the DSB repair machinery results in cells utilising remaining factors to effect repair, but often at the cost of increased mutagenesis. Here we discuss the complex regulation of the Immediate-Early, Early, and Late responses to DSBs proceeding repair itself.

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XRCC1 protects transcription from toxic PARP1 activity during DNA base excision repair

Cited by 38 publications

References 92 publications

PARP1 and XRCC1 exhibit a reciprocal relationship in genotoxic stress response

PARP1 and XRCC1 exhibit a reciprocal relationship in genotoxic stress response

XRCC1 counteracts poly(ADP ribose)polymerase (PARP) poisons, olaparib and talazoparib, and a clinical alkylating agent, temozolomide, by promoting the removal of trapped PARP1 from broken DNA

Immediate-Early, Early, and Late Responses to DNA Double Stranded Breaks

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