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Chromatin Regulates Genome Targeting with Cisplatin |
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| Authors: | Dr Emmanouil Zacharioudakis Poonam Agarwal Dr Alexandra Bartoli Nathan Abell Lavaniya Kunalingam Dr Valérie Bergoglio Dr Blerta Xhemalce Dr Kyle M Miller Dr Raphaël Rodriguez |
| Institution: | 1. Institut Curie, PSL Research University, Chemical Cell Biology Group, 26 Rue d'Ulm, 75248 Paris Cedex 05, France
CNRS UMR3666, 75005 Paris, France INSERM U1143, 75005 Paris, France Institut de Chimie des Substances Naturelles, UPR2301, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France These authors contributed equally to this work.;2. Department of Molecular Biosciences, Institute of Cellular and Molecular Biology, University of Texas at Austin, 2506 Speedway Stop A5000, Austin, TX, 78712 USA These authors contributed equally to this work.;3. Institut Curie, PSL Research University, Chemical Cell Biology Group, 26 Rue d'Ulm, 75248 Paris Cedex 05, France CNRS UMR3666, 75005 Paris, France INSERM U1143, 75005 Paris, France Institut de Chimie des Substances Naturelles, UPR2301, 1 Avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France;4. Department of Molecular Biosciences, Institute of Cellular and Molecular Biology, University of Texas at Austin, 2506 Speedway Stop A5000, Austin, TX, 78712 USA;5. CRCT, University of Toulouse, INSERM, CNRS, UPS, Avenue Hubert Curien, 31037 Toulouse, France;6. Institut Curie, PSL Research University, Chemical Cell Biology Group, 26 Rue d'Ulm, 75248 Paris Cedex 05, France |
| Abstract: | Cisplatin derivatives can form various types of DNA lesions (DNA-Pt) and trigger pleiotropic DNA damage responses. Here, we report a strategy to visualize DNA-Pt with high resolution, taking advantage of a novel azide-containing derivative of cisplatin we named APPA, a cellular pre-extraction protocol and the labeling of DNA-Pt by means of click chemistry in cells. Our investigation revealed that pretreating cells with the histone deacetylase (HDAC) inhibitor SAHA led to detectable clusters of DNA-Pt that colocalized with the ubiquitin ligase RAD18 and the replication protein PCNA. Consistent with activation of translesion synthesis (TLS) under these conditions, SAHA and cisplatin cotreatment promoted focal accumulation of the low-fidelity polymerase Polη that also colocalized with PCNA. Remarkably, these cotreatments synergistically triggered mono-ubiquitination of PCNA and apoptosis in a RAD18-dependent manner. Our data provide evidence for a role of chromatin in regulating genome targeting with cisplatin derivatives and associated cellular responses. |
| Keywords: | Chromatin Cisplatin Histon-Deacetylase Klickchemie Krebs |