title: Structural insights into the recognition of cisplatin and AAF-dG lesion by Rad14 (XPA).
abstract: Nucleotide excision repair (NER) is responsible for the removal of a large variety of structurally diverse DNA lesions. Mutations of the involved proteins cause the xeroderma pigmentosum (XP) cancer predisposition syndrome. Although the general mechanism of the NER process is well studied, the function of the XPA protein, which is of central importance for successful NER, has remained enigmatic. It is known, that XPA binds kinked DNA structures and that it interacts also with DNA duplexes containing certain lesions, but the mechanism of interactions is unknown. Here we present two crystal structures of the DNA binding domain (DBD) of the yeast XPA homolog Rad14 bound to DNA with either a cisplatin lesion (1,2-GG) or an acetylaminofluorene adduct (AAF-dG). In the structures, we see that two Rad14 molecules bind to the duplex, which induces DNA melting of the duplex remote from the lesion. Each monomer interrogates the duplex with a β-hairpin, which creates a 13mer duplex recognition motif additionally characterized by a sharp 70° DNA kink at the position of the lesion. Although the 1,2-GG lesion stabilizes the kink due to the covalent fixation of the crosslinked dG bases at a 90° angle, the AAF-dG fully intercalates into the duplex to stabilize the kinked structure.
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: Sandra C
authors.initials: SC
authors.lastname: Koch
authors.affiliation: Rudolf Virchow Center for Experimental Biomedicine, Institute for Structural Biology, University of Würzburg, 97080 Wurzburg, Germany;
authors.forename: Jochen
authors.initials: J
authors.lastname: Kuper
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: Karola L
authors.initials: KL
authors.lastname: Gasteiger
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: Nina
authors.initials: N
authors.lastname: Simon
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: Ralf
authors.initials: R
authors.lastname: Strasser
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: David
authors.initials: D
authors.lastname: Eisen
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany;
authors.forename: Simon
authors.initials: S
authors.lastname: Geiger
authors.affiliation: Department of Chemistry, Technische Universität München, 85747 Garching, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Schneider
authors.affiliation: Rudolf Virchow Center for Experimental Biomedicine, Institute for Structural Biology, University of Würzburg, 97080 Wurzburg, Germany; thomas.carell@cup.uni-muenchen.de caroline.kisker@virchow.uni-wuerzburg.de.
authors.forename: Caroline
authors.initials: C
authors.lastname: Kisker
authors.affiliation: Center for Integrated Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, 81377 Munich, Germany; thomas.carell@cup.uni-muenchen.de caroline.kisker@virchow.uni-wuerzburg.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Carell
mesh_terms: D015073:2-Acetylaminofluorene / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D002945:Cisplatin / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004249:DNA Damage; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013816:Thermodynamics; D044366:Transition Temperature
chemical_list: D004271:DNA, Fungal; C072975:RAD14 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015073:2-Acetylaminofluorene; D045643:DNA Repair Enzymes; D002945:Cisplatin
keywords: AAF; NER; Rad14; XPA; cisplatin