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. Stephen.Pettitt@icr.ac.uk.
11 Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. Stephen.Pettitt@icr.ac.uk.
12 Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. Andrew.Tutt@icr.ac.uk.
13 MRC Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, Oxford, UK. Kristijan.Ramadan@oncology.ox.ac.uk.
14 The CRUK Gene Function Laboratory, London, UK. Chris.Lord@icr.ac.uk.
15 Breast Cancer Now Toby Robins Research Centre, The Institute of Cancer Research, London, UK. Chris.Lord@icr.ac.uk.
# 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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