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Nucleic Acids Research, 1984, Vol. 12, No. 3 1725-1736
© 1984

MOLECULAR BIOLOGY

Osmium tetroxide: a new probe for site-specific distortions in supercoiled DNAs

Gerardo C. Glikin*,+, Marie Vojtískova, Liliana Rena-Descalzi* and Emil Palecek§

Institute of Biophysics, Czechoslovak Academy of Sciences 612 65 Brno, Czechoslovakia, USA *Department of Biology, University of Rochester Rochester, NY 14627, USA

§To whom correspondence should be addressed

Received September 28, 1983. Accepted December 19, 1983.

Supercoiled plasmids Col El and cDm 506 (a Col El derivative carrying the D. melanogaster histone gene repeat) were treated with 0s04 in presence of pyriaine and the reaction products were analyzed using different approaches. Gel electrophoresis showed that 0s04 binding to supercoiled DNA induced its relaxation without nicking. The amount of osmium bound to DNA (as determined electrochernjcally) increased with the extent of DNA relaxation. As a result of osmium modification of supercoiled cDm 506, a single denaturation "bubble" was observed in the electron microscope. Mapping of the osmium binding site by S1 nuclease cleavage followed by restriction enzyme digestion has revealed one major site in the intergenic spacer between the H1 and H3 histone genes of D. melanogaster. This site differs from the site cleaved by S1 nuclease in supercoiled DNA in the absence of osmium.

+ Present Address: Institut de Recherches Scientifiques sur le Cancer, Villejuif Cedex 94802, France


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