title: Characterization of a human DNA damage binding protein implicated in xeroderma pigmentosum E.
abstract: A human DNA damage binding protein implicated in the DNA excision repair disorder xeroderma pigmentosum E was purified to near homogeneity from HeLa cells. The protein is abundant (approximately 10(5) copies/cell) and has a native molecular weight of 154,000-163,000 as estimated by gel filtration and glycerol gradient sedimentation. DNA damage binding activity copurified with polypeptides of 124 and 41 kDa. Based on the native molecular weight, cosedimentation of both polypeptides with DNA damage binding activity on glycerol gradients, and a molar ratio of approximately 1:1 for the two polypeptides, it appears that p124 and p41 are subunits of a heterodimeric protein. Binding to damaged DNA was resistant to K+ concentrations approaching 1 M, but showed anion-specific sensitivity to Cl- concentrations above 0.5 M, suggesting that the majority of the binding energy is contributed by nonionic interactions. In contrast to previous reports, the DNA damage binding protein was shown to recognize cyclobutane pyrimidine dimers in addition to a nonphotoreactivable lesion(s), most likely the pyrimidine-pyrimidone (6-4) photoproduct.
authors.affiliation: Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720.
authors.forename: S
authors.initials: S
authors.lastname: Keeney
authors.forename: G J
authors.initials: GJ
authors.lastname: Chang
authors.forename: S
authors.initials: S
authors.lastname: Linn
mesh_terms: D002850:Chromatography, Gel; D004249:DNA Damage / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D013379:Substrate Specificity; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins