The ubiquitin-dependent ATPase p97 removes cytotoxic trapped PARP1 from chromatin

  • Nat Cell Biol. 2022 Jan;24(1):62-73. doi: 10.1038/s41556-021-00807-6.
Dragomir B Krastev  #  1  2 ,  Shudong Li  #  3 ,  Yilun Sun  #  4 ,  Andrew J Wicks  1  2 ,  Gwendoline Hoslett  3 ,  Daniel Weekes  2 ,  Luned M Badder  5 ,  Eleanor G Knight  2 ,  Rebecca Marlow  2 ,  Mercedes Calvo Pardo  6 ,  Lu Yu  6 ,  Tanaji T Talele  7 ,  Jiri Bartek  8  9 ,  Jyoti S Choudhary  6 ,  Yves Pommier  4 ,  Stephen J Pettitt  10  11 ,  Andrew N J Tutt  12 ,  Kristijan Ramadan  13 ,  Christopher J Lord  14  15
Affiliations
  • 1. The CRUK Gene Function Laboratory, London, UK.
  • 2. Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK.
  • 3. MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford, UK.
  • 4. Developmental Therapeutics Branch, Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA.
  • 5. The Breast Cancer Now Research Unit, King's College London, London, UK.
  • 6. Functional Proteomics Laboratory, The Institute of Cancer Research, London, UK.
  • 7. Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, NY, USA.
  • 8. Danish Cancer Society Research Center, Copenhagen, Denmark.
  • 9. Division of Genome Biology, Department of Medical Biochemistry and Biophysics, Science for Life Laboratory, Karolinska Institute, Stockholm, Sweden.
  • 10. The CRUK Gene Function Laboratory, London, UK. [email protected].
  • 11. Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. [email protected].
  • 12. Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. [email protected].
  • 13. MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford, UK. [email protected].
  • 14. The CRUK Gene Function Laboratory, London, UK. [email protected].
  • 15. Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. [email protected].
  • # Contributed equally.
PMID: 35013556 DOI: 10.1038/s41556-021-00807-6
Abstract

Poly (ADP-ribose) polymerase (PARP) inhibitors elicit antitumour activity in homologous recombination-defective cancers by trapping PARP1 in a chromatin-bound state. How cells process trapped PARP1 remains unclear. Using wild-type and a trapping-deficient PARP1 mutant combined with rapid immunoprecipitation mass spectrometry of endogenous proteins and Apex2 proximity labelling, we delineated mass spectrometry-based interactomes of trapped and non-trapped PARP1. These analyses identified an interaction between trapped PARP1 and the ubiquitin-regulated p97 ATPase/segregase. We found that following trapping, PARP1 is SUMOylated by PIAS4 and subsequently ubiquitylated by the SUMO-targeted E3 ubiquitin Ligase RNF4, events that promote recruitment of p97 and removal of trapped PARP1 from chromatin. Small-molecule p97-complex inhibitors, including a metabolite of the clinically used drug disulfiram (CuET), prolonged PARP1 trapping and enhanced PARP inhibitor-induced cytotoxicity in homologous recombination-defective tumour cells and patient-derived tumour organoids. Together, these results suggest that p97 ATPase plays a key role in the processing of trapped PARP1 and the response of tumour cells to PARP inhibitors.