Biopolym. Cell. 1991; 7(2):8-15.
Structure and Function of Biopolymers
Fractionation of eukaryotic DNA by pulsed field gel electrophoresis. Enzymatic nature of the formation
1Solovyan V. T., 1Makytryuck V. L., 1Rudenko G. N.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Institute of Cell Biology and Genetic Engineering, NAS of Ukraine
    148, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680

Abstract

The data presented here confirm the possibility of enzymatic formation of discrete DNA-fragments, appearing during the fractionation of lysed nuclei by FIGE. Teniposide-dependent pattern of DNA-fragments and occurrence of protein-linked DNA breaks suggest that discrete cleavage of intact nuclear DNA is modulated by DNA-topoisomerase II. It is pointed that pulsed field gel electrophoresis may be a useful method to study higer-order chromatin structure.

References

[1] Solovyan VT, Kunakh VA. The fractionation of eukaryotic DNA by the field inversion gel electrophoresis. 1. Detection of discrete fragments. Biopolym Cell. 1990; 6(3):97-9.
[2] Solovyan VT. The fractionation of eukaryotic DNA by field inversion gel electrophoresis. properties of discrete fragments. Biopolym Cell. 1991; 7(1):85-7.
[3] Razin SV, Mant'eva VL, Georgiev GP. Isolation and comparative characteristics of the DNA attached to the axial structures of interphase nuclei and metaphase chromosomes. Mol Biol (Mosk). 1980;14(1):223-33. Russian.
[4] Burton K. Preparation of apurinic acid and of oligodeoxyribonucleotides with formic acid and diphenylamine. Methods in Enzymology ; 1967;222–4.
[5] Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.
[6] Yoshitake S, Yamada Y, Ishikawa E, Masseyeff R. Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl)-maleimide. Eur J Biochem. 1979;101(2):395-9.
[7] Yang L, Rowe TC, Liu LF. Identification of DNA topoisomerase II as an intracellular target of antitumor epipodophyllotoxins in simian virus 40-infected monkey cells. Cancer Res. 1985;45(11 Pt 2):5872-6.
[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.
[9] Paulson JR, Laemmli UK. The structure of histone-depleted metaphase chromosomes. Cell. 1977;12(3):817-28.
[10] 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.
[11] Blasquez VC, Sperry AO, Garrard WT. Elements that organize chromosomal loops in the interphase nucleus. DNA-protein interactions in transcriptio. New York : Alan R. Liss, 1989:273-86.
[12] 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.