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Nucleic Acids Research, 1992, Vol. 20, No. 8 1973-1977
© 1992

MOLECULAR BIOLOGY

Eukaryotic topoisomerase II cleavage of parallel stranded DNA tetraplexes

I.K. Chung, V.B. Mehta, J.R. Spitzner{dagger} and M.T. Muller*

Department of Molecular Genetics, The Ohio State University Columbus, OH 43210, USA

*To whom correspondence should be addressed

Received December 20, 1991. Revised March 19, 1992. Accepted March 19, 1992.

A guanine-rich single-stranded DNA from the human immunoglobulin switch region was shown by Sen and Gilbert [Nature, (1988) 334, 364–366] to be able to self-associate to form a stable four-stranded parallel DNA structure. Topoisomerase II did not cleave the single-stranded DNA molecule. Surprisingly, the enzyme did cleave the same DNA sequence when it was annealed into the four-stranded structure. The two cleavage sites observed were the same as those found when this DNA molecule was paired with a complementary molecule to create a normal B-DNA duplex. These cleavages were shown to be protein-linked and reversible by the addition of salt, suggesting a normal topolsomerase II reaction mechanism. In addition, an eight-stranded DNA molecule created by the association of a complementary oligonucleotide with the four-stranded structure was also cleaved by topolsomerase II despite being resistant to restriction endonuclease digestion. These results suggest that a single strand of DNA may possess the sequence Information to direct topoisomerase II to a binding site, but the site must be base paired in a proper manner to do so. This demonstration of the ability of a four-stranded DNA molecule to be a substrate for an enzyme further suggests that these DNA structures may be present in cells.

{dagger}Present address: Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA


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