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Nucleic Acids Research, 1991, Vol. 19, No. 11 2861-2868
© 1991

MOLECULAR BIOLOGY

Sequence requirements for mammalian topoisomerase II mediated DNA cleavage stimulated by an ellipticine derivative

Philippe Fosse, Brigitte Rene, Marc Le Bret, Claude Paoletti and Jean-Marie Saucier

Laboratoire de Biochimie-Enzymologie, CNRS URA 147, INSERM U 140, Institut Gustave Roussy 94805 Villejuif cedex, France

Received March 20, 1991. Revised April 30, 1991. Accepted April 30, 1991.

Various antitumor drugs stabilize DNA topoisomerase II-DNA transient covalent complexes. The complexes distribution along pBR322 DNA was shown previously to depend upon the nature of the drug (Tewey et al. (1984) Science 226, 466–468). The position in pBR322 of DNA cleavage by calf DNA topoisomerase II for 115 such sites stabilized by an ellipticine derivative and the relative frequency of cleavage at most of these sites were determined. The nucleotide sequence surrounding the 25 strongest sites was analyzed and the following ellipticine specific consensus sequence was deduced: 5'-ANCNT(A/G)T:NN(G/C)N(A/G)-3' where cleavage occurs at the indicated mark. A thymine is always present at the 3' end of at least one strand of the strong cleavage sites, and the dinucleotide AT or GT at the 3' end of the break plays a major role in the complex stabilisation. The predictive value of cleavage of the consensus was tested for two regions of SV40 DNA and cleavage was indeed detected at the majority of the sites matching the consensus. Some complexes stabilized by ellipticine are resistant to salt dissociation and this property seems to be correlated with the presence of symmetrical sequences in the cleavage site with a center of symmetry staggered relatively to the center of symmetry of cleavage.


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