Complex of Osmium Tetroxide with 1,10-Phenanthroline Binds Covalently to Double-Stranded DNA

Abstract

Complex of osmium tetroxide with 1,10–phenanthroline (Os, phen) reacts with double-stranded B-DNA in contrast to osmium tetroxide, pyridine and other osmium structural probes which show a strong preference for single-stranded DNA (ssDNA) (Palecek, E. in Abelson, J.N., and Simon, M.I. (eds), Lilley, D.M.J., and Dahlberg, J.E., (volume eds.), Methods in Enzymology, Vol. 212, DNA Structures, part B., Academic Press, 139–155 (1992)). Modification of negatively supercoiled DNA (scDNA) with Os, phen changes the DNA electrophoretic mobility inducing the DNA relaxation at lower degrees of modification followed by formation of positive supercoils at higher modification extents. Electrophoretic mobility of the Os, phen-modified DNA fragments in agarose gel is almost unchanged while a strong retardation of the same fragments is observed in Polyacrylamide gels. Os, phen- modified DNA is hypersensitive to nuclease S1. Cleavage of this DNA by restriction enzymes is selectively inhibited showing a preference of Os.phen for TA and AT dinu-cleotide steps. DNA modification by Os.phen is inhibited by low and moderate concentrations of MgCl₂. The covalent binding of Os.phen to double-stranded DNA (dsDNA) is preceded by noncovalent interactions (probably intercalation) inducing DNA structural changes; the shape of the Os, phen-modified DNA molecule appears to be severely deformed.