title: Effects of spectator ligands on the specific recognition of intrastrand platinum-DNA cross-links by high mobility group box and TATA-binding proteins.
abstract: The results presented describe the effects of various spectator ligands, attached to a platinum 1,2-intrastand d(GpG) cross-link in duplex DNA, on the binding of high mobility group box (HMGB) domains and the TATA-binding protein (TBP). In addition to cisplatin-modified DNA, 15-base pair DNA probes modified by [Pt(1R,2R-diaminocyclohexane)](2+), cis-[Pt(NH(3))(cyclohexylamine)](2+), [Pt(ethylenediamine)](2+), cis-[Pt(NH(3))(cyclobutylamine)](2+), and cis-[Pt(NH(3))(2-picoline)](2+) were examined. Electrophoretic mobility shift assays show that both the A and B domains of HMGB1 as well as TBP discriminate between different platinum-DNA adducts. HMGB1 domain A is the most sensitive to the nature of the spectator ligands on platinum. The effect of the spectator ligands on protein binding also depends highly on the base pairs flanking the platinated d(GpG) site. Double-stranded oligonucleotides containing the AG*G*C sequence, where the asterisks denote the sites of platination, with different spectator ligands are only moderately discriminated by the HMGB proteins and TBP, but the recognition of dsTG*G*A is highly dependent on the ligands. The effects of HMGB1 overexpression in a BG-1 ovarian cancer cell line, induced by steroid hormones, on the sensitivity of cells treated with [Pt(1R,2R-diaminocyclohexane)Cl(2)] and cis-[Pt(NH(3))(cyclohexylamine)Cl(2)] were also examined. The results suggest that HMGB1 protein levels influence the cellular processing of cis-[Pt(NH(3))- (cyclohexylamine)](2+), but not [Pt((1R,2R)-diaminocyclohexane)](2+), DNA lesions. This result is consistent with the observed binding of HMGB1a to platinum-modified dsTG*G*A probes but not with the binding affinity of HMGB1a and HMGB1 to platinum-damaged dsAG*G*C oligonucleotides. These experiments reinforce the importance of sequence context in platinum-DNA lesion recognition by cellular proteins.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
authors.forename: M
authors.initials: M
authors.lastname: Wei
authors.forename: S M
authors.initials: SM
authors.lastname: Cohen
authors.forename: A P
authors.initials: AP
authors.lastname: Silverman
authors.forename: S J
authors.initials: SJ
authors.lastname: Lippard
mesh_terms: D002945:Cisplatin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008956:Models, Chemical; D015345:Oligonucleotide Probes / Q000494:pharmacology; D010051:Ovarian Neoplasms / Q000378:metabolism; D010984:Platinum / Q000494:pharmacology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D008024:Ligands; D015345:Oligonucleotide Probes; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; D010984:Platinum; D004247:DNA; D002945:Cisplatin