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Nucleic Acids Research, 1985, Vol. 13, No. 4 1135-1149
© 1985

Articles

DNAaseI-hypersensitive minichromosomes of SV40 possess an elastic torsional strain in DNA

Andrey N. Luchnik*, Valerii V. Bakayev1, Anatolii A. Yugai1, Ilyia B. Zbarsky and Georgii P. Georgiev1

Institute of Developmental Biology, USSR Academy of Sciences Vavilov str. 26 1Institute of Molecular Biology, USSR Academy of Sciences Vavilov str 32, Moscow 117334, USSR

To whom reprint requests should be sent

Received December 12, 1984. Accepted January 24, 1985.

Previously, we have shown that DNA in a small fraction (2–5%) of SV40 minichromosomes was torsionally strained and could be relaxed by treating minichromosomes with topoisomerase I. This fraction was enriched with endogoneous RNA polymerase II (Luchnik et al 1982, EMBO J., 1, 1353). Here we show that one and the same fraction of SV4O 1 minichromosomes is hypersensitive to DNAase I and is relaxable by topoisomerase I. Moreover, this fraction completely loses its hypersensitivity to 1 DNAase I upon relaxation. The possibility that this fraction of minichromosomes can be represented by naked DNA is ruled out by the results of studying the kinetics of minichromosome digestion by DNAase I in comparison to digestion of pure SV40 DNA and by measuring the buoyant density of SV40 chromatin in equilibrium CaCl gradient. Our data obtained with SV4O minichromosomes may be relevant to the mechanism responsible for DNAase I hypersensitivity in the loops or domains of cellular chromatin.


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