Skip to main content

Purification of the Bacteriophage T4 Type II DNA Topoisomerase

  • Protocol
  • First Online:
DNA Topoisomerase Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 94))

Abstract

Bacteriophage T4 encodes a type II topoisomerase with properties more similar to those of the eukaryotic class of enzymes than to those of the bacterial DNA gyrase (1,2). Indeed, the discovery of the T4 topoisomerase provided the first example of an ATP-dependent relaxing enzyme (3,4), and an understanding of the properties of the T4 enzyme rapidly led to the discovery of similar enzymes from eukaryotic cells (5-7).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Kreuzer, K. N. and Jongeneel, C. V. (1983) Escherichia coli phage T4 topoisomerase. Methods Enzymol. 100, 144–160.

    Article  CAS  Google Scholar 

  2. Wang, J. C. (1985) DNA topoisomerases. Annu. Rev. Biochem. 54, 665–697.

    Article  CAS  Google Scholar 

  3. Liu, L. F., Liu, C. C., and Alberts, B. M. (1979) T4 DNA topoisomerase: a new ATP-dependent enzyme essential for initiation of T4 bacteriophage DNA replication. Nature 281, 456–461.

    Article  CAS  Google Scholar 

  4. Stetler, G. L., King, G. J., and Huang, W. M. (1979) T4 DNA-delay proteins, required for specific DNA replication, form a complex that has ATP-dependent DNA topoisomerase activity. Proc. Natl. Acad. Sci. USA 76, 3737–3741.

    Article  CAS  Google Scholar 

  5. Liu, L. F., Liu, C. C., and Alberts, B. M. (1980) Type II DNA topoisomerases: Enzymes that can unknot a topologically knotted DNA molecule via a reversible double-strand break. Cell 19, 697–707.

    Article  CAS  Google Scholar 

  6. Hsieh, T.-S. and Brutlag, D. (1980) ATP-dependent DNA topoisomerase from D. melanogaster reversibly catenates duplex DNA rings. Cell 21, 115–125.

    Article  CAS  Google Scholar 

  7. Baldi, M. I., Benedetti, P., Mattoccia, E., and Tocchini-Valentini, G. P. (1980) In vitro catenation and decatenation of DNA and a novel eucaryotic ATP-dependent topoisomerase. Cell 20, 461–467.

    Article  CAS  Google Scholar 

  8. Kreuzer, K. N. (1994) A bacteriophage model system for studying topoisomerase inhibitors. Adv. Pharmacol. 29B, 171–186.

    Article  CAS  Google Scholar 

  9. Kreuzer, K. N. (1989) DNA topoisomerases as potential targets of antiviral action. Pharmacol. Ther. 43, 377–395.

    Article  CAS  Google Scholar 

  10. Huff, A. C. and Kreuzer, K. N. (1991) The mechanism of antitumor drug action in a simple bacteriophage model system, in DNA Topoisomerases in Cancer (Potmesil M., and Kohn, K. W.), Oxford University Press, NY, pp. 215–229.

    Google Scholar 

  11. Rowe, T. C., Tewey, K. M., and Liu, L. F. (1984) Identification of the breakagereunion subunit of T4 DNA topoisomerase. J. Biol. Chem. 259, 9177–9181.

    Article  CAS  Google Scholar 

  12. Huff, A. C. and Kreuzer, K. N. (1990) Evidence for a common mechanism of action for antitumor and antibacterial agents that inhibit type II DNA topoisomerases. J. Biol. Chem. 265, 20,496–20,505.

    Article  CAS  Google Scholar 

  13. Freudenreich, C. H. and Kreuzer, K. N. (1993) Mutational analysis of a type II topoisomerase cleavage site: distinct requirements for enzyme and inhibitors. EMBO J. 12, 2085–2097.

    Article  CAS  Google Scholar 

  14. Huff, A. C., Leatherwood, J. K., and Kreuzer, K. N. (1989) Bacteriophage T4 DNA topoisomerase is the target of antitumor agent 4′-(9-acridinylamino) methanesulfon-m-anisidide (m-AMSA) in T4-infected Escherichia coli. Proc. Natl. Acad. Sci. USA 86, 1307–1311.

    Article  CAS  Google Scholar 

  15. Huff, A. C., Ward, R. E., IV, and Kreuzer, K. N. (1990) Mutational alteration of the breakage/resealing subunit of bacteriophage T4 DNA topoisomerase confers resistance to antitumor agent m-AMSA. Mol. Gen. Genet. 221, 27–32.

    Article  CAS  Google Scholar 

  16. Morris, C. F., Hana-Inaba, H., Mace, D., Sinha, N. K., and Alberts, B. (1979) Purification of the gene 43, 44, 45, and 62 proteins of the bacteriophage T4 DNA replication apparatus. J. Biol. Chem. 254, 6787–6796.

    Article  CAS  Google Scholar 

  17. Karam, J. D. (1994) Molecular Biology of Bacteriophage T4. ASM, Washington, DC.

    Google Scholar 

  18. Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    Article  CAS  Google Scholar 

  19. Singer, B. S. and Gold, L. (1991) Phage T4 expression vector: protection from proteolysis. Gene 106, 1–6.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Humana Press Inc.

About this protocol

Cite this protocol

Kreuzer, K.N., Neece, S.H. (1999). Purification of the Bacteriophage T4 Type II DNA Topoisomerase. In: Bjornsti, MA., Osheroff, N. (eds) DNA Topoisomerase Protocols. Methods in Molecular Biology, vol 94. Humana, Totowa, NJ. https://doi.org/10.1385/1-59259-259-7:171

Download citation

  • DOI: https://doi.org/10.1385/1-59259-259-7:171

  • Published:

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-0-89603-444-0

  • Online ISBN: 978-1-59259-259-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics