PMID:7673159
| Citation |
Lambert, B, Jestin, JL, Bréhin, P, Oleykowski, C, Yeung, AT, Mailliet, P, Prétot, C, Le Pecq, JB, Jacquemin-Sablon, A and Chottard, JC (1995) Binding of the Escherichia coli UvrAB proteins to the DNA mono- and diadducts of cis-[N-2-amino-N-2-methylamino-2,2,1-bicycloheptane]dichloroplatinum(II ) and cisplatin. Analysis of the factors controlling recognition and proof of monoadduct-mediated UvrB-DNA cross-linking. J. Biol. Chem. 270:21251-7 |
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| Abstract |
The interactions of the Escherichia coli endonuclease UvrAB proteins with the DNA mono- and diadducts of both the cis-racemic exo-[N-2-amino-N-2-methylamino-2,2,1-bicycloheptane]dichloroplatin um(II) (complex 1) and cisplatin (cis-diamminedichloroplatinum(II) (cis-DDP)), have been studied. Complex 1 reacts faster with DNA than cis-DDP and gives monoadducts with a longer lifetime (8 h 20 min chelation t 1/2 compared with 2 h 40 min for cis-DDP). Using pSP65 plasmid [3H]DNA, the filter binding assay was associated with the analysis of the nucleoprotein complexes to characterize the UvrAB recognition of the platinum adducts and to demonstrate the occurrence of platinum-mediated DNA-protein cross-linking. First, it is shown that the UvrAB proteins recognize the complex 1 mono- and diadducts with a higher affinity than those of cis-DDP. Fifteen times more cis-DDP adducts per plasmid are required than complex 1 adducts, to lead to similar UvrAB binding. However, the UvrAB proteins recognize monoadducts and diadducts of each complex with a similar affinity. Second, it is shown that UvrB is the protein involved in the nucleo-protein complexes formed from mono- and diadducts of complex 1 and cis-DDP. This protein is also partly cross-linked to DNA with a similar efficiency by monoadducts derived from complex 1 and cis-DDP. However, as UvrB has a greater affinity for the DNA adducts of complex 1 than for those of cis-DDP, more UvrB-platinum-DNA cross-links are formed with complex 1 than with cis-DDP. This study, using a bacterial repair system as a model, points to a possible strategy for making new cytotoxic platinum complexes for mammalian cells. |
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| Keywords |
Adenosine Triphosphatases/metabolism; Bacterial Proteins/metabolism; Chelating Agents; Cisplatin/metabolism; Cross-Linking Reagents; DNA Adducts/metabolism; DNA Helicases; DNA, Bacterial/metabolism; DNA-Binding Proteins/metabolism; Edetic Acid; Escherichia coli/metabolism; Escherichia coli Proteins; Kinetics; Organoplatinum Compounds/metabolism; Protein Binding; Sodium Dodecyl Sulfate |
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