Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

CHK2-mediated regulation of PARP1 in oxidative DNA damage response

Abstract

Poly(ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor, which upon activation, recruits downstream proteins by poly(ADP-ribosyl)ation (PARylation). However, it remains largely unclear how PARP1 activity is regulated. Interestingly, the data obtained through this study revealed that PARP1 was co-immunoprecipitated with checkpoint kinase 2 (CHK2), and the interaction was increased after oxidative DNA damage. Moreover, CHK2 depletion resulted in a reduction in overall PARylation. To further explore the functional relationship between PARP1 and CHK2, this study employed H2O2 to induce an oxidative DNA damage response in cells. Here, we showed that CHK2 and PARP1 interact in vitro and in vivo through the CHK2 SCD domain and the PARP1 BRCT domain. Furthermore, CHK2 stimulates the PARylation activity of PARP1 through CHK2-dependent phosphorylation. Consequently, the impaired repair associated with PARP1 depletion could be rescued by re-expression of wild-type PARP1 and the phospho-mimic but not the phospho-deficient mutant. Mechanistically, we showed that CHK2-dependent phosphorylation of PARP1 not only regulates its cellular localization but also promotes its catalytic activity and its interaction with XRCC1. These findings indicate that CHK2 exerts a multifaceted impact on PARP1 in response to oxidative stress to facilitate DNA repair and to maintain cell survival.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  1. Panieri E, Santoro MM. ROS homeostasis and metabolism: a dangerous liason in cancer cells. Cell Death Dis. 2016;7:e2253.

    Article  CAS  Google Scholar 

  2. Hegde ML, Izumi T, Mitra S. Oxidized base damage and single-strand break repair in mammalian genomes: role of disordered regions and posttranslational modifications in early enzymes. Prog Mol Biol Transl Sci. 2012;110:123–53.

    Article  CAS  Google Scholar 

  3. Noren Hooten N, Kompaniez K, Barnes J, Lohani A, Evans MK. Poly(ADP-ribose) polymerase 1 (PARP-1) binds to 8-oxoguanine-DNA glycosylase (OGG1). J Biol Chem. 2011;286:44679–90.

    Article  Google Scholar 

  4. Barkauskaite E, Jankevicius G, Ahel I. Structures and mechanisms of enzymes employed in the synthesis and degradation of PARP-dependent protein ADP-ribosylation. Mol Cell. 2015;58:935–46.

    Article  CAS  Google Scholar 

  5. Pascal JM, Ellenberger T. The rise and fall of poly(ADP-ribose): an enzymatic perspective. DNA Repair (Amst). 2015;32:10–16.

    Article  CAS  Google Scholar 

  6. Perina D, Mikoc A, Ahel J, Cetkovic H, Zaja R, Ahel I. Distribution of protein poly(ADP-ribosyl)ation systems across all domains of life. DNA Repair (Amst). 2014;23:4–16.

    Article  CAS  Google Scholar 

  7. Kim MY, Mauro S, Gévry N, Lis JT, Kraus WL. NAD+-dependent modulation of chromatin structure and transcription by nucleosome binding properties of PARP-1. Cell. 2004;119:803–14.

    Article  CAS  Google Scholar 

  8. Altmeyer M, Neelsen KJ, Teloni F, Pozdnyakova I, Pellegrino S, Grofte M, et al. Liquid demixing of intrinsically disordered proteins is seeded by poly(ADP-ribose). Nat Commun. 2015;6:8088.

    Article  CAS  Google Scholar 

  9. Muthurajan UM, Hepler MR, Hieb AR, Clark NJ, Kramer M, Yao T, et al. Automodification switches PARP-1 function from chromatin architectural protein to histone chaperone. Proc Natl Acad Sci USA. 2014;111:12752–7.

    Article  CAS  Google Scholar 

  10. Vyas S, Matic I, Uchima L, Rood J, Zaja R, Hay RT, et al. Family-wide analysis of poly(ADP-ribose) polymerase activity. Nat Commun. 2014;5:4426.

    Article  CAS  Google Scholar 

  11. Du Y, Yamaguchi H, Wei Y, Hsu JL, Wang HL, Hsu YH, et al. Blocking c-Met-mediated PARP1 phosphorylation enhances anti-tumor effects of PARP inhibitors. Nat Med. 2016;22:194–201.

    Article  CAS  Google Scholar 

  12. Gagné JP, Moreel X, Gagne P, Labelle Y, Droit A, Chevalier-Pare M, et al. Proteomic investigation of phosphorylation sites in poly(ADP-ribose) polymerase-1 and poly(ADP-ribose) glycohydrolase. J Proteome Res. 2009;8:1014–29.

    Article  Google Scholar 

  13. Gibson BA, Kraus WL. New insights into the molecular and cellular functions of poly(ADP-ribose) and PARPs. Nat Rev Mol Cell Biol. 2012;13:411–24.

    Article  CAS  Google Scholar 

  14. Hassa PO, Haenni SS, Buerki C, Meier NI, Lane WS, Owen H, et al. Acetylation of poly(ADP-ribose) polymerase-1 by p300/CREB-binding protein regulates coactivation of NF-kappaB-dependent transcription. J Biol Chem. 2005;280:40450–64.

    Article  CAS  Google Scholar 

  15. Messner S, Schuermann D, Altmeyer M, Kassner I, Schmidt D, Schar P, et al. Sumoylation of poly(ADP-ribose) polymerase 1 inhibits its acetylation and restrains transcriptional coactivator function. FASEB J. 2009;23:3978–89.

    Article  CAS  Google Scholar 

  16. Martin N, Schwamborn K, Schreiber V, Werner A, Guillier C, Zhang XD, et al. PARP-1 transcriptional activity is regulated by sumoylation upon heat shock. EMBO J. 2009;28:3534–48.

    Article  CAS  Google Scholar 

  17. Ryu H, Al-Ani G, Deckert K, Kirkpatrick D, Gygi SP, Dasso M, et al. PIASy mediates SUMO-2/3 conjugation of poly(ADP-ribose) polymerase 1 (PARP1) on mitotic chromosomes. J Biol Chem. 2010;285:14415–23.

    Article  CAS  Google Scholar 

  18. Zhang S, Lin Y, Kim YS, Hande MP, Liu ZG, Shen HM. c-Jun N-terminal kinase mediates hydrogen peroxide-induced cell death via sustained poly(ADP-ribose) polymerase-1 activation. Cell Death Differ. 2007;14:1001–10.

    Article  CAS  Google Scholar 

  19. Meehan RS, Chen AP. New treatment option for ovarian cancer: PARP inhibitors. Gynecol Oncol Res Pract. 2016;3:3.

    Article  Google Scholar 

  20. Rouleau M, Patel A, Hendzel MJ, Kaufmann SH, Poirier GG. PARP inhibition: PARP1 and beyond. Nat Rev Cancer. 2010;10:293–301.

    Article  CAS  Google Scholar 

  21. Vyas S, Chang P. New PARP targets for cancer therapy. Nat Rev Cancer. 2014;14:502–9.

    Article  CAS  Google Scholar 

  22. Helleday T. The underlying mechanism for the PARP and BRCA synthetic lethality: clearing up the misunderstandings. Mol Oncol. 2011;5:387–93.

    Article  CAS  Google Scholar 

  23. Hu Y, Petit SA, Ficarro SB, Toomire KJ, Xie A, Lim E, et al. PARP1-driven poly-ADP-ribosylation regulates BRCA1 function in homologous recombination-mediated DNA repair. Cancer Discov. 2014;4:1430–47.

    Article  CAS  Google Scholar 

  24. Ray Chaudhuri A, Callen E, Ding X, Gogola E, Duarte AA, Lee JE, et al. Replication fork stability confers chemoresistance in BRCA-deficient cells. Nature. 2016;535:382–7.

    Article  Google Scholar 

  25. Sonnenblick A, de Azambuja E, Azim HA Jr, Piccart M. An update on PARP inhibitors--moving to the adjuvant setting. Nat Rev Clin Oncol. 2015;12:27–41.

    Article  CAS  Google Scholar 

  26. Zhang F, Shi J, Bian C, Yu X. Poly(ADP-ribose) mediates the BRCA2-dependent early DNA damage response. Cell Rep. 2015;13:678–89.

    Article  Google Scholar 

  27. Chou WC, Hu LY, Hsiung CN, Shen CY. Initiation of the ATM-Chk2 DNA damage response through the base excision repair pathway. Carcinogenesis. 2015;36:832–40.

    Article  CAS  Google Scholar 

  28. Zannini L, Delia D, Buscemi G. CHK2 kinase in the DNA damage response and beyond. J Mol Cell Biol. 2014;6:442–57.

    Article  CAS  Google Scholar 

  29. Zhao H, Traganos F, Albino AP, Darzynkiewicz Z. Oxidative stress induces cell cycle-dependent Mre11 recruitment, ATM and Chk2 activation and histone H2AX phosphorylation. Cell Cycle. 2008;7:1490–5.

    Article  CAS  Google Scholar 

  30. Ahn JY, Li X, Davis HL, Canman CE. Phosphorylation of threonine 68 promotes oligomerization and autophosphorylation of the Chk2 protein kinase via the forkhead-associated domain. J Biol Chem. 2002;277:19389–95.

    Article  CAS  Google Scholar 

  31. Xu X, Tsvetkov LM, Stern DF. Chk2 activation and phosphorylation-dependent oligomerization. Mol Cell Biol. 2002;22:4419–32.

    Article  CAS  Google Scholar 

  32. Chou WC, Wang HC, Wong FH, Ding SL, Wu PE, Shieh SY, et al. Chk2-dependent phosphorylation of XRCC1 in the DNA damage response promotes base excision repair. EMBO J. 2008;27:3140–50.

    Article  CAS  Google Scholar 

  33. Tan Y, Raychaudhuri P, Costa RH. Chk2 mediates stabilization of the FoxM1 transcription factor to stimulate expression of DNA repair genes. Mol Cell Biol. 2007;27:1007–16.

    Article  CAS  Google Scholar 

  34. Yeh YH, Huang YF, Lin TY, Shieh SY. The cell cycle checkpoint kinase CHK2 mediates DNA damage-induced stabilization of TTK/hMps1. Oncogene. 2009;28:1366–78.

    Article  CAS  Google Scholar 

  35. Kauppinen TM, Chan WY, Suh SW, Wiggins AK, Huang EJ, Swanson RA. Direct phosphorylation and regulation of poly(ADP-ribose) polymerase-1 by extracellular signal-regulated kinases 1/2. Proc Natl Acad Sci USA. 2006;103:7136–41.

    Article  CAS  Google Scholar 

  36. Matsuoka S, Ballif BA, Smogorzewska A, McDonald ER 3rd, Hurov KE, Luo J, et al. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage. Science. 2007;316:1160–6.

    Article  CAS  Google Scholar 

  37. Wright RH, Castellano G, Bonet J, Le Dily F, Font-Mateu J, Ballare C, et al. CDK2-dependent activation of PARP-1 is required for hormonal gene regulation in breast cancer cells. Genes Dev. 2012;26:1972–83.

    Article  CAS  Google Scholar 

  38. Brunyanszki A, Olah G, Coletta C, Szczesny B, Szabo C. Regulation of mitochondrial poly(ADP-Ribose) polymerase activation by the beta-adrenoceptor/cAMP/protein kinase A axis during oxidative stress. Mol Pharmacol. 2014;86:450–62.

    Article  Google Scholar 

  39. Shang F, Zhang J, Li Z, Yin Y, Wang Y, Marin TL, et al. Cardiovascular protective effect of metformin and telmisartan: reduction of PARP1 activity via the AMPK-PARP1 cascade. PLoS ONE. 2016;11:e0151845.

    Article  Google Scholar 

  40. Ray Chaudhuri A, Nussenzweig A. The multifaceted roles of PARP1 in DNA repair and chromatin remodelling. Nat Rev Mol Cell Biol. 2017;18:610–21.

    Article  CAS  Google Scholar 

  41. El-Khamisy SF, Masutani M, Suzuki H, Caldecott KW. A requirement for PARP-1 for the assembly or stability of XRCC1 nuclear foci at sites of oxidative DNA damage. Nucleic Acids Res. 2003;31:5526–33.

    Article  CAS  Google Scholar 

  42. Stolz A, Ertych N, Bastians H. Tumor suppressor CHK2: regulator of DNA damage response and mediator of chromosomal stability. Clin Cancer Res. 2011;17:401–5.

    Article  CAS  Google Scholar 

  43. Bauer PI, Farkas G, Buday L, Mikala G, Meszaros G, Kun E, et al. Inhibition of DNA binding by the phosphorylation of poly ADP-ribose polymerase protein catalysed by protein kinase C. Biochem Biophys Res Commun. 1992;187:730–6.

    Article  CAS  Google Scholar 

  44. Langelier MF, Planck JL, Roy S, Pascal JM. Structural basis for DNA damage-dependent poly(ADP-ribosyl)ation by human PARP-1. Science. 2012;336:728–32.

    Article  CAS  Google Scholar 

  45. Mansoorabadi SO, Wu M, Tao Z, Gao P, Pingali SV, Guo L, et al. Conformational activation of poly(ADP-ribose) polymerase-1 upon DNA binding revealed by small-angle X-ray scattering. Biochemistry. 2014;53:1779–88.

    Article  CAS  Google Scholar 

  46. Breslin C, Hornyak P, Ridley A, Rulten SL, Hanzlikova H, Oliver AW, et al. The XRCC1 phosphate-binding pocket binds poly (ADP-ribose) and is required for XRCC1 function. Nucleic Acids Res. 2015;43:6934–44.

    Article  CAS  Google Scholar 

  47. Li M, Lu LY, Yang CY, Wang S, Yu X. The FHA and BRCT domains recognize ADP-ribosylation during DNA damage response. Genes Dev. 2013;27:1752–68.

    Article  CAS  Google Scholar 

  48. Loeffler PA, Cuneo MJ, Mueller GA, DeRose EF, Gabel SA, London RE. Structural studies of the PARP-1 BRCT domain. BMC Struct Biol. 2011;11:37.

    Article  CAS  Google Scholar 

  49. Pleschke JM, Kleczkowska HE, Strohm M, Althaus FR. Poly(ADP-ribose) binds to specific domains in DNA damage checkpoint proteins. J Biol Chem. 2000;275:40974–80.

    Article  CAS  Google Scholar 

  50. Bahassi el M, Myer DL, McKenney RJ, Hennigan RF, Stambrook PJ. Priming phosphorylation of Chk2 by polo-like kinase 3 (Plk3) mediates its full activation by ATM and a downstream checkpoint in response to DNA damage. Mutat Res. 2006;596:166–76.

    Article  CAS  Google Scholar 

  51. Buscemi G, Carlessi L, Zannini L, Lisanti S, Fontanella E, Canevari S, et al. DNA damage-induced cell cycle regulation and function of novel Chk2 phosphoresidues. Mol Cell Biol. 2006;26:7832–45.

    Article  CAS  Google Scholar 

  52. Anderson VE, Walton MI, Eve PD, Boxall KJ, Antoni L, Caldwell JJ, et al. CCT241533 is a potent and selective inhibitor of CHK2 that potentiates the cytotoxicity of PARP inhibitors. Cancer Res. 2011;71:463–72.

    Article  CAS  Google Scholar 

  53. Lord CJ, Ashworth A. BRCAness revisited. Nat Rev Cancer. 2016;16:110–20.

    Article  CAS  Google Scholar 

  54. Bouwman P, Aly A, Escandell JM, Pieterse M, Bartkova J, van der Gulden H, et al. 53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers. Nat Struct Mol Biol. 2010;17:688–95.

    Article  CAS  Google Scholar 

  55. Hong R, Ma F, Zhang W, Yu X, Li Q, Luo Y, et al. 53BP1 depletion causes PARP inhibitor resistance in ATM-deficient breast cancer cells. BMC Cancer. 2016;16:725.

    Article  Google Scholar 

  56. Lord CJ, Ashworth A. Mechanisms of resistance to therapies targeting BRCA-mutant cancers. Nat Med. 2013;19:1381–8.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to the Proteomic Core at Institute of Biomedical Sciences for the assistance with the identification of the PARP1 phosphorylation sites. This work was supported by funding from Academia Sinica, Taiwan to S-YS.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sheau-Yann Shieh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hsu, PC., Gopinath, R.K., Hsueh, YA. et al. CHK2-mediated regulation of PARP1 in oxidative DNA damage response. Oncogene 38, 1166–1182 (2019). https://doi.org/10.1038/s41388-018-0506-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-018-0506-7

This article is cited by

Search

Quick links