Fractionation of eukaryotic DNA by pulsed field gel electrophoresis. Enzymatic nature of the formation

Authors

  • V. T. Solovyan Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author
  • V. L. Makytryuck Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author
  • G. N. Rudenko Institute of Molecular Biology and Genetics, NAS of Ukraine 150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680 Author

DOI:

https://doi.org/10.7124/bc.0002BC

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

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.

Solovyan VT. The fractionation of eukaryotic DNA by field inversion gel electrophoresis. properties of discrete fragments. Biopolym Cell. 1991; 7(1):85-7.

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.

Burton K. Preparation of apurinic acid and of oligodeoxyribonucleotides with formic acid and diphenylamine. Methods in Enzymology ; 1967;222–4.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5.

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.

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.

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.

Paulson JR, Laemmli UK. The structure of histone-depleted metaphase chromosomes. Cell. 1977;12(3):817-28.

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.

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.

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.

Published

1991-03-20

Issue

Section

Structure and Function of Biopolymers