PMID:8756462

Citation	Zamble, DB, Mu, D, Reardon, JT, Sancar, A and Lippard, SJ (1996) Repair of cisplatin--DNA adducts by the mammalian excision nuclease. Biochemistry 35:10004-13
Abstract	Nucleotide excision repair is one of the many cellular defense mechanisms against the toxic effects of cisplatin. An in vitro excision repair assay employing mammalian cell-free extracts was used to determine that the 1,2-d(ApG) intrastrand cross-link, a prevalent cisplatin-DNA adduct, is excised by the excinuclease from a site-specifically modified oligonucleotide 156 base pairs in length. Repair of the minor interstrand d(G)/d(G) cross-link was not detected by using this system. Proteins containing the high mobility group (HMG) domain DNA-binding motif, in particular, rat HMG1 and a murine testis-specific HMG-domain protein, specifically inhibit excision repair of the intrastrand 1,2-d(GpG) and -d(ApG) cross-links. This effect was also exhibited by a single HMG domain from HMG1. Similar inhibition of repair of a site-specific 1,2-d(GpG) intrastrand cross-link by an HMG-domain protein also occurred in a reconstituted system containing highly purified repair factors. These results indicate that HMG-domain proteins can block excision repair of the major cisplatin-DNA adducts and suggest that such an activity could contribute to the unique sensitivity of certain tumors to the drug. The reconstituted excinuclease was more efficient at excising the 1,3-d(GpTpG) intrastrand adduct than either the 1,2-d(GpG) or d(ApG) intrastrand adducts, in agreement with previous experiments using whole cell extracts [Huang, J. -C., Zamble, D. B., Reardon, J. T., Lippard, S. J., Sancar, A. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 10394-10398]. This result suggests that structural differences among the platinated DNA substrates, and not the presence of unidentified cellular factors, determine the relative excision repair rates of cisplatin-DNA intrastrand cross-links in the whole cell extracts.
Links	PubMed Online version:10.1021/bi960453+
Keywords	Animals; Base Sequence; Binding Sites; Cholesterol/metabolism; Cisplatin; DNA Adducts; DNA Repair; Endodeoxyribonucleases/metabolism; HeLa Cells; High Mobility Group Proteins/chemistry; High Mobility Group Proteins/metabolism; High Mobility Group Proteins/pharmacology; Humans; Kinetics; Male; Mammals; Mice; Molecular Sequence Data; Oligodeoxyribonucleotides; Rats; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Recombinant Proteins/pharmacology; Substrate Specificity; Testis
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