Biopolym. Cell. 1993; 9(6):52-59.
Structure and Function of Biopolymers
Fractionation of eukaryotic DNA by pulsed field gel electrophoresis. The peculiarities of topoisomerase / DNA complex behaviour in isolated nuclei
- Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
The properties of topoisomerase II/DNA complex in nuclear preparations have been investigated. It has been shown that topoII/nuclear DNA complex functiones unitypically both in Mg2+- and in Ca2+-containing media. Unlike to the in vitro assay the treatment of topoII/nuclear DNA complex with EDTA resulted in increase of nuclear DNA cleavage rather than its rejoining. The nuclear topoII/DNA complex treated with SDS was found to acquire the capacity to subsequent SDS-independent nuclear DNA cleavage. The salient features of topoisomerase II/nuclear DNA complex are supposed to be due to the topoII enzyme involvement in higher level's chromatin compactization leading to the initial stabilization of topoII/nuclear DNA complex in vivo conditions. The availability in cells the different topoII/DNA complex variants of possible physiological value has been proposed.
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 Solovyan VT, Andreyev IO, Kunakh VA. The fractionation of eukaryotic dna by pulsed field gel electrophoresis. Evidence for nuclear DNA organization as constituent component of topoisomerase II/DNA complex. Biopolym Cell. 1993; 9(5):44-51.
 Solov?an VT, Kunakh VA. Fractionation of eukaryotic DNA in a pulsating electrical field. I. Detection and properties of discrete DNA fragments. Mol Biol (Mosk). 1991;25(4):1071-9.
 Solov'ian VT, Andreev IO, Kunakh VA. Fractionation of eukaryotic DNA in a pulsed electrical field. II. Discrete DNA fragments and level of structural organization of chromatin. Mol Biol (Mosk). 1991;25(6):1483-91.
 Sander M, Hsieh T. Double strand DNA cleavage by type II DNA topoisomerase from Drosophila melanogaster. J Biol Chem. 1983;258(13):8421-8.
 Liu LF, Rowe TC, Yang L, Tewey KM, Chen GL. Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem. 1983;258(24):15365-70.
 Osheroff N, Zechiedrich EL. Calcium-promoted DNA cleavage by eukaryotic topoisomerase II: trapping the covalent enzyme-DNA complex in an active form. Biochemistry. 1987;26(14):4303-9.
 Osheroff N. Effect of antineoplastic agents on the DNA cleavage/religation reaction of eukaryotic topoisomerase II: inhibition of DNA religation by etoposide. Biochemistry. 1989;28(15):6157-60.
 Earnshaw WC, Heck MM. Localization of topoisomerase II in mitotic chromosomes. J Cell Biol. 1985;100(5):1716-25.
 Gasser SM, Laemmli UK. The organisation of chromatin loops: characterization of a scaffold attachment site. EMBO J. 1986;5(3):511-8.
 Berrios M, Osheroff N, Fisher PA. In situ localization of DNA topoisomerase II, a major polypeptide component of the Drosophila nuclear matrix fraction. Proc Natl Acad Sci U S A. 1985;82(12):4142-6.
 Cockerill PN, Garrard WT. Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites. Cell. 1986;44(2):273-82.
 Earnshaw WC, Halligan B, Cooke CA, Heck MM, Liu LF. Topoisomerase II is a structural component of mitotic chromosome scaffolds. J Cell Biol. 1985;100(5):1706-15.
 Uemura T, Ohkura H, Adachi Y, Morino K, Shiozaki K, Yanagida M. DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe. Cell. 1987;50(6):917-25.
 Newport J. Nuclear reconstitution in vitro: stages of assembly around protein-free DNA. Cell. 1987;48(2):205-17.
 Sperry AO, Blasquez VC, Garrard WT. Dysfunction of chromosomal loop attachment sites: illegitimate recombination linked to matrix association regions and topoisomerase II. Proc Natl Acad Sci U S A. 1989;86(14):5497-501.
 Adachi Y, K?s E, Laemmli UK. Preferential, cooperative binding of DNA topoisomerase II to scaffold-associated regions. EMBO J. 1989;8(13):3997-4006.