title: Comparative analysis of binding of human damaged DNA-binding protein (XPE) and Escherichia coli damage recognition protein (UvrA) to the major ultraviolet photoproducts: T[c,s]T, T[t,s]T, T[6-4]T, and T[Dewar]T.
abstract: Human cells contain a protein that binds to UV-irradiated DNA with high affinity. This protein, the damaged DNA-binding protein (DDB), is absent from some xeroderma pigmentosum complementation group E cell strains; therefore, it has been suggested that it may be the damage recognition subunit of a human excision nuclease complex. However, the identity of the UV photoproduct bound by DDB and the role of this protein in nucleotide excision repair have been controversial. In this study, we used several synthetic DNA substrates, each of which contains one of the major UV photoproducts, and DDB purified to apparent homogeneity to quantify the specific binding of DDB to various photoproducts. For comparison, the binding of the same photoproducts by the Escherichia coli damage recognition protein UvrA, which is known to be a subunit of the E. coli excision nuclease, was also measured. UvrA and DDB each bound with high affinity to T[t,s]T, T[6-4]T, and T[Dewar]T, but only marginally discriminated between an undamaged oligomer and an oligomer with a T[c,s]T. In contrast to these similarities with regard to the binding to UV photoproducts, UvrA bound to another excision repair substrate, the psoralen-thymine monoadduct, with high specificity, whereas DDB was unable to distinguish between psoralen-adducted DNA and undamaged DNA. We conclude that DDB may play a special role in the repair of UV damage, but it cannot be the sole damage recognition subunit of human excision nuclease.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill 27599.
authors.forename: J T
authors.initials: JT
authors.lastname: Reardon
authors.forename: A F
authors.initials: AF
authors.lastname: Nichols
authors.forename: S
authors.initials: S
authors.lastname: Keeney
authors.forename: C A
authors.initials: CA
authors.lastname: Smith
authors.forename: J S
authors.initials: JS
authors.lastname: Taylor
authors.forename: S
authors.initials: S
authors.lastname: Linn
authors.forename: A
authors.initials: A
authors.lastname: Sancar
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000138:chemical synthesis / Q000378:metabolism* / Q000528:radiation effects; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D011564:Furocoumarins / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D010777:Photochemistry; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D011564:Furocoumarins; D004247:DNA; C068631:UvrA protein, E coli; D000251:Adenosine Triphosphatases