title: Platinum anticancer drug damage enforces a particular rotational setting of DNA in nucleosomes.
abstract: We constructed two site-specifically modified nucleosomes containing an intrastrand cis-{Pt(NH3)2}2+ 1,3-d(GpTpG) cross-link, similar to one formed by the anticancer drugs carboplatin and cisplatin on DNA, and investigated their structures by hydroxyl radical footprinting and exonuclease III digestion. Hydroxyl radical footprinting demonstrated that the presence of the platinum cross-link selects out a specific rotational setting of DNA on the histone octamer core in each of two reconstituted nucleosomes in which the platinum positions differ by half a DNA helical turn. The {Pt(NH3)2}2+ cross-link is situated in a structurally similar location, with the undamaged strand projecting outward, forcing the DNA to adopt opposite rotational settings in its wrapping around the histone octamer in the two nucleosomes. Enzymatic digestion by exonuclease III of the nucleosome substrates revealed that the platinum cross-link affects the translational positioning of the DNA, forcing it into an asymmetric arrangement with respect to the core histone proteins. We suggest that these phasing phenomena may be central to the recognition and processing of platinum-DNA adducts in cancer cells treated with these drugs and possibly may be common to other DNA damaging events.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Danford
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
authors.forename: Qun
authors.initials: Q
authors.lastname: Wang
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Tullius
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Lippard
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016466:CHO Cells; D016190:Carboplatin / Q000494:pharmacology*; D002945:Cisplatin / Q000494:pharmacology*; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018736:DNA Adducts / Q000737:chemistry*; D004249:DNA Damage; D018983:DNA Footprinting; D004260:DNA Repair; D005090:Exodeoxyribonucleases; D017665:Hydroxyl Radical / Q000737:chemistry; D066298:In Vitro Techniques; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry* / Q000187:drug effects*
chemical_list: D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D018736:DNA Adducts; D009707:Nucleosomes; D017665:Hydroxyl Radical; D016190:Carboplatin; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III; D002945:Cisplatin