title: TOX4 and its binding partners recognize DNA adducts generated by platinum anticancer drugs.
abstract: 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.
authors.affiliation: CEA Grenoble, INAC, SCIB (UMR E_3 CEA-Université Joseph Fourier, CNRS FRE3200)-Laboratoire Lésions des Acides Nucléiques, 17 Rue des Martyrs, 38054 Grenoble Cedex 09, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Bounaix Morand du Puch
authors.forename: Ewa
authors.initials: E
authors.lastname: Barbier
authors.forename: Alexandra
authors.initials: A
authors.lastname: Kraut
authors.forename: Yohann
authors.initials: Y
authors.lastname: Couté
authors.forename: Julia
authors.initials: J
authors.lastname: Fuchs
authors.forename: Arnaud
authors.initials: A
authors.lastname: Buhot
authors.forename: Thierry
authors.initials: T
authors.lastname: Livache
authors.forename: Michel
authors.initials: M
authors.lastname: Sève
authors.forename: Alain
authors.initials: A
authors.lastname: Favier
authors.forename: Thierry
authors.initials: T
authors.lastname: Douki
authors.forename: Didier
authors.initials: D
authors.lastname: Gasparutto
authors.forename: Sylvie
authors.initials: S
authors.lastname: Sauvaigo
authors.forename: Jean
authors.initials: J
authors.lastname: Breton
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry*; D002467:Cell Nucleus / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008280:Magnetics; D009363:Neoplasm Proteins / Q000378:metabolism*; D009944:Organoplatinum Compounds / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D040901:Proteomics; D020349:Surface Plasmon Resonance
chemical_list: D000970:Antineoplastic Agents; D018736:DNA Adducts; D008024:Ligands; D009363:Neoplasm Proteins; D009944:Organoplatinum Compounds; C482285:TOX4 protein, human