Abstract
DNA topoisomerases I and II (topo I and topo II) are ubiquitous nuclear enzymes that regulate DNA topology by breaking and resealing one or both strands of a DNA duplex. Topoisomerase I makes a single-stranded break in a DNA duplex, mediates passage of the intact strand through the break, and then reseals it. Type II topoisomerases create transient breaks in both strands of a duplex, pass an intact DNA segment through that break, and then reseal the cleavage site. This ability of topoisomerases to catalyze concerted breaking and rejoining of DNA strands makes them biologically important enzymes that are involved in crucial cellular processes (1–3).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Vosberg H. P. (1985) DNA topoisomerases: Enzymes that control DNA conformation. Curr. Top. Microbiol. Immunol. 114, 19–102.
Wang. J. C. (1985) DNA topoisomerases. Annu. Rev. Biochem. 54, 665–697.
Osheroff N. (1989) Biochemical basis for the interactions of type I and type II topoisomerases with DNA. Pharmacol. Ther. 41,223–241.
Trask D. K., DiDonato J. A., and Muller M. T. (1984) Rapid detection and isolation of covalent DNA/protein complexes: application to topoisomerase I and II. EMBOJ. 3, 671–676.
Gromova I. I., Kjeldsen E., Svejstrup J. Q., Alsner J., Christiansen K., and Westergaard O. (1993) Characterization of an altered catalysis of a camptothecin-resistant eukaryotic topoisomerase I. Nucleic Acids Res. 21,593–600.
Morham S. G. and Shuman S. (1992) Covalent and noncovalent DNA binding by mutants of vaccinia DNA topoisomerase I. J. Biol. Chem. 267,15,984–15,992.
D’Arpa P. and Liu L. F. (1989) Topoisomerase-targeting antitumor drugs. Biochem. Biophys. Acta. 989,163–177.
Schneider E., Hsiang Y. H., and Liu L. F. 1990. DNA topoisomerases as anti-cancer drug targets. Pharmacology 21, 149–183.
Zwelling L. A. (1985) DNA topoisomerase II as a target of antineoplastic drug therapy. Cancer Metastasis Rev. 4, 263–276.
Glisson B. S. and Ross W. E. (1987) DNA topoisomerase II: A primer on the enzyme and its unique role as a multidrug target in cancer chemotherapy. Pharmacol. Ther. 32, 89–106.
Liu L. F. (1989) DNA topoisomerase poisons as antitumor drugs. Annu. Rev. Biochem. 58,351–375.
Covey J. M., Jaxel C., Kohn K., and Pommier Y. (1989) Protein-linked DNA strand breaks induced in mammalian cells by camptothecin, an inhibitor of topoisomerase I. Cancer Res. 49,5016–5022.
Hertzberg R. P., Caranfa M. J., Holden K. G., Jakas D. R., Gallagher G., Mattern M. R., Mong S. M., Bartus J. O., Johnson R. K., and Kingsbury W. D. (1989) Modification of the hydroxy lactone ring of camptothecin: inhibition of mammalian topoisomerase I and biological activity. J. Med. Chem. 32,715–720.
Burke T. G., Mishra W. K., Wani M. C., and Wall M. E. (1993) Lipid bilayer partitioning and stability of camptothecin drugs. Biochemistry 32,5352–5364.
Muller M. T., Pfund W. P., Mehta V. B., and Trask D. K. (1985) Eukaryotic type I topoisomerase is enriched in the nucleolus and catalytically active on ribo-somal DNA. EMBO J. 4, 1237–1243.
Trask D. K. and Muller M. T. (1988) Stabilization of type I topoisomerase-DNA covalent complexes by actinomycin D. Proc. Natl. Acad. Sci. USA 85,1417–1421.
Shaw J. L., Blanco J., and Mueller G. C. (1974) A simple procedure for isolation of DNA, RNA, and protein fractions from cultured animal cells. Anal. Biochem. 65, 125–131.
Subramanian D., Kraut E., Staubus A., Young D. C., and Muller M. T. (1995) Analysis of topoisomerase I-DNA complexes in patients administered topotecan. Cancer Res. 55, 2097–2103.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Subramanian, D., Furbee, C.S., Muller, M.T. (2001). ICE Bioassay. In: Osheroff, N., Bjornsti, MA. (eds) DNA Topoisomerase Protocols. Methods in Molecular Biology™, vol 95. Humana Press. https://doi.org/10.1385/1-59259-057-8:137
Download citation
DOI: https://doi.org/10.1385/1-59259-057-8:137
Publisher Name: Humana Press
Print ISBN: 978-0-89603-512-6
Online ISBN: 978-1-59259-057-5
eBook Packages: Springer Protocols