Uvr excision repair protein complex of Escherichia coli binds to the convex side of a cisplatin-induced kink in the DNA.
The Escherichia coli UvrABC endonuclease is capable of initiating the repair of a wide variety of DNA damages. To study the binding of the UvrAB complex to the DNA at the site of a lesion we have constructed a synthetic DNA fragment with a defined cis-diamminedichloroplatinum(II) (cis-Pt).GG adduct. The cis-Pt.GG is the major adduct after treatment of DNA with the antitumor agent cisplatin. Binding to the DNA at the site of the defined lesion was studied with DNase I and MPE.Fe(II) hydroxyl radical footprinting. The results indicate that the UvrAB complex binds to the convex side of the kink in the DNA caused by the cis-Pt.GG adduct. Concerted incisions of the damaged strand by the UvrABC endonuclease were at the 8th phosphodiester bond 5' to and at the 4th bond 3' of the adjacent guanines. An additional incision was found at the 15th phosphodiester bond 5' to the damaged site. This extra incision was stimulated by a high concentration of UvrC.
Visse
R
R
Department of Biochemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
de Ruijter
M
M
Brouwer
J
J
Brandsma
J A
JA
van de Putte
P
P
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D002945:Cisplatin / Q000494:pharmacology*; D016172:DNA Fingerprinting; D004260:DNA Repair; D004269:DNA, Bacterial / Q000187:drug effects* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D006878:Hydroxides; D017665:Hydroxyl Radical; D008969:Molecular Sequence Data
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D006878:Hydroxides; D017665:Hydroxyl Radical; D004706:Endodeoxyribonucleases; C036385:endodeoxyribonuclease uvrABC; D002945:Cisplatin