TOX4 and its binding partners recognize DNA adducts generated by platinum anticancer drugs

• Published in: Archives of biochemistry and biophysics Vol. 507; no. 2; pp. 296 - 303
• Main Authors: Puch, Christophe Bounaix Morand du, Barbier, Ewa, Kraut, Alexandra, Couté, Yohann, Fuchs, Julia, Buhot, Arnaud, Livache, Thierry, Sève, Michel, Favier, Alain, Douki, Thierry, Gasparutto, Didier, Sauvaigo, Sylvie, Breton, Jean
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.03.2011; Elsevier
• Subjects: Antineoplastic Agents - chemistry; Biotechnology; Cell Nucleus - metabolism; chromatin; cisplatin; cytotoxicity; DNA adducts; DNA Adducts - chemistry; DNA Adducts - genetics; DNA Adducts - metabolism; DNA damage; DNA lesions; DNA repair; drug therapy; extracts; HeLa Cells; Humans; image analysis; Life Sciences; Ligands; Magnetics; Neoplasm Proteins - metabolism; Organoplatinum Compounds - chemistry; pharmacology; plasmids; Plasmids - genetics; platinum; Platinum anticancer drugs; PNUTS; Protein Binding; Proteomics; SPRi; Surface Plasmon Resonance; tandem mass spectrometry; TOX4; WDR82; TOX4; Platinum anticancer drugs; SPRi; PNUTS; DNA lesions; WDR82
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/j.abb.2010.12.021

► TOX4/PNUTS/WDR82 complex interacts with DNA damaged by anticancer platinum drugs. ► SPRi shows that TOX4 affinity for platinum adducts is weaker than that of HMGB1. ► Plasmid probes and proteomics allow identification of damaged DNA interactome. Platinating agents are commonly prescribed anticancer drugs damaging DNA. Induced lesions are recognized by a wide range of proteins. These are involved in cellular mechanisms such as DNA repair, mediation of cytotoxicity or chromatin remodeling. They therefore constitute crucial actors to understand pharmacology of these drugs. To expand our knowledge about this subproteome, we developed a ligand fishing trap coupled to high throughput proteomic tools. This trap is made of damaged plasmids attached to magnetic beads, and was exposed to cell nuclear extracts. Retained proteins were identified by nanoHPLC coupled to tandem mass spectrometry. This approach allowed us to establish a list of 38 proteins interacting with DNA adducts generated by cisplatin, oxaliplatin and satraplatin. Some of them were already known interactome members like high mobility group protein 1 (HMGB1) or the human upstream binding factor (hUBF), but we also succeeded in identifying unexpected proteins such as TOX HMG box family member 4 (TOX4), phosphatase 1 nuclear targeting subunit (PNUTS), and WD repeat-containing protein 82 (WDR82), members of a recently discovered complex. Interaction between TOX4 and platinated DNA was subsequently validated by surface plasmon resonance imaging (SPRi). These interactions highlight new cellular responses to DNA damage induced by chemotherapeutic agents.