title: Interactions between UvrA and UvrB: the role of UvrB's domain 2 in nucleotide excision repair.
abstract: Nucleotide excision repair (NER) is a highly conserved DNA repair mechanism present in all kingdoms of life. UvrB is a central component of the bacterial NER system, participating in damage recognition, strand excision and repair synthesis. None of the three presently available crystal structures of UvrB has defined the structure of domain 2, which is critical for the interaction with UvrA. We have solved the crystal structure of the UvrB Y96A variant, which reveals a new fold for domain 2 and identifies highly conserved residues located on its surface. These residues are restricted to the face of UvrB important for DNA binding and may be critical for the interaction of UvrB with UvrA. We have mutated these residues to study their role in the incision reaction, formation of the pre-incision complex, destabilization of short duplex regions in DNA, binding to UvrA and ATP hydrolysis. Based on the structural and biochemical data, we conclude that domain 2 is required for a productive UvrA-UvrB interaction, which is a pre-requisite for all subsequent steps in nucleotide excision repair.
authors.affiliation: Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Truglio
authors.forename: Deborah L
authors.initials: DL
authors.lastname: Croteau
authors.forename: Milan
authors.initials: M
authors.lastname: Skorvaga
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: DellaVecchia
authors.forename: Karsten
authors.initials: K
authors.lastname: Theis
authors.forename: Bhaskar S
authors.initials: BS
authors.lastname: Mandavilli
authors.forename: Bennett
authors.initials: B
authors.lastname: Van Houten
authors.forename: Caroline
authors.initials: C
authors.lastname: Kisker
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D001407:Bacillus / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002850:Chromatography, Gel; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D004249:DNA Damage; D004265:DNA Helicases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D024202:Electrophoretic Mobility Shift Assay; D020558:GTP Phosphohydrolases / Q000378:metabolism; D014644:Genetic Variation; D006860:Hydrogen Bonding; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017354:Point Mutation; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013059:Spectrum Analysis, Raman; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D000251:Adenosine Triphosphatases; D020558:GTP Phosphohydrolases; D004265:DNA Helicases