2. Academic Validation
  3. The ubiquitin-dependent ATPase p97 removes cytotoxic trapped PARP1 from chromatin

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

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.

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