title: Strand-specific binding of RPA and XPA to damaged duplex DNA.
abstract: The nucleotide excision repair (NER) pathway is a major pathway used to repair bulky adduct DNA damage. Two proteins, xeroderma pigmentosum group A protein (XPA) and replication protein A (RPA), have been implicated in the role of DNA damage recognition in the NER pathway. The particular manner in which these two damage recognition proteins align themselves with respect to a damaged DNA site was assessed using photoreactive base analogues within specific DNA substrates to allow site-specific cross-linking of the damage recognition proteins. Results of these studies demonstrate that both RPA and XPA are in close proximity to the adduct as measured by cross-linking of each protein directly to the platinum moiety. Additional studies demonstrate that XPA contacts both the damaged and undamaged strands of the duplex DNA. Direct evidence is presented demonstrating preferential binding of RPA to the undamaged strand of a duplex damaged DNA molecule.
authors.affiliation: Department of Biochemistry and Molecular Biology, Wright State University School of Medicine, Dayton, Ohio 45435, USA.
authors.forename: Ingrid L
authors.initials: IL
authors.lastname: Hermanson-Miller
authors.forename: John J
authors.initials: JJ
authors.lastname: Turchi
mesh_terms: D002945:Cisplatin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry* / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry; D011485:Protein Binding; D051716:Replication Protein A; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics; D051760:Xeroderma Pigmentosum Group A Protein
chemical_list: D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009692:Nucleic Acid Heteroduplexes; D009841:Oligonucleotides; D051716:Replication Protein A; D051760:Xeroderma Pigmentosum Group A Protein; D002945:Cisplatin