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Nucleic Acids Research, 1985, Vol. 13, No. 20 7395-7412
© 1985

Articles

Reactions of the UVRABC excision nuclease with DNA damaged by diamminedichloroplatinum(II)

D.J. Beck1, S. Popoff1, A. Sancar2 and W.D. Rupp3

1Department of Biological Sciences, Bowling Green State University Bowling Green, OH 43403 2University of North Carolina School of Medicine Chapel Hill, NC 27514 3Yale University School of Medicine New Haven, CT 06520, USA

Received June 5, 1985. Revised September 26, 1985. Accepted September 26, 1985.

Mutants of Escherichia coil, which are blocked in excision repair (uvrA6, uvrB5, or uvrC34) are exceptionally sensitive to the antitumor drug cis-Pt(II) (NH3)2Cl2 (cis-DDP) but not to the trans isomer. Plasmid DNA, damaged by either the cis or trans compound and treated with the UVRABC excision nuclease was cut as shown by conversion of supercoiled DNA to relaxed forms. All three protein products of the uvrA, uvrB, and uvrC genes were required for incision. End-labeled fragments damaged with cis-DDP and reacted with the UVRABC nuclease were cut at the 8th phosphodiester bond 5' and at the 4th phosphodiester bond 3' to adjacent GG's. DNA treated with trans-DDP was not cut appreciably at adjacent GG's by the repair enzyme as subsequent analysis of reaction products after enzyme digestion gave a pattern similar to those obtained with control untreated fragments. The results indicate that the UVRABC nuclease may promote cell survival by the removal of adjacent GG's which are crosslinked by cis-Pt(II)(NH3)2Cl2.


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