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Nucleic Acids Research, 1995, Vol. 23, No. 13 2381-2388
© 1995

MOLECULAR BIOLOGY

Intrastrand cross-links are not formed in the reaction between transplatin and native DNA: relationm with the clinical inefficency of transplatin

Marc Boudvillain, Rozenn Dalbiès, Christophe Aussourd and Marc Leng*

Centre de Biophysique Molecularie CBNRS, Rue Charles Sadron, 45071 Orleans Cedex 2, France

*To whom corespondence should be addressed

Received April 20, 1995. Accepted May 19, 1995.

The reaction between trans-dlammlnedichloroplatinum( ll) and single-stranded oligonucleotides containing the sequence d(GXG) (X being an adenlne, cytosine or thymlne residue) yields trans-(Pt(NH3)2[d(GXG)-GN7,GN7]} Intrastrand cross-links. These cross-links do not prevent the pairing of the platinated oligonucleotides with their complementary strands but they decrease the thermal stability of the duplexes. The thermal stability is not much affected by the chemical nature of the X residue and Its complementary base. By gel electrophoresis, it is shown that the trans- {Pt(NH3)2[d(GTG)-GN7,GN7]} cross-link bends the DNA double helix (26°) and unwinds It (45°). The pairing of the platinated oligonucleotides with their complementary strands promotes the rearrangement f the 1,3-intrastrand cross-links into Interstrand cross-links. At a given temperature, the nature of the X residue, its complementary base and of the base pairs adjacent to the adducts do not dramatically ffect the ate of the reaction. To know whether trans-{Pt(NH3)2[d(GXG)-GN7,GN7]} cross-links do not rearrange In some sequences, the location of these adducts was searched In double-stranded DNA after reaction with frans-dlamminedichloroplatinum(ll) by means of the 3'–5' exonuclease activity of T4 DNA polymerase. At low level of platination, trans-{Pt(NH3}2[d(GXG}-GN7,GN7]} cross-links were not detected. Monofunctional adducts and interstrand cross-links were mainly formed. These results are discussed in relation with the clinical Inefficiency of trans- iammlnedlchloroplatinum(II).


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