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Nucleic Acids Research, 1983, Vol. 11, No. 12 4019-4034
© 1983

MOLECULAR BIOLOGY

Reversible decatenation of kinetoplast DNA by a DNA topoisomerase from trypanosomatids

Joseph Shlomai and Anat Zadok

The S.F. Kuvin Centre for the Study of Tropical and Infectious Diseases The Hebrew University-Hadassah Medical School, Jerusalem 91010, Israel

Received March 29, 1983. Revised May 20, 1983. Accepted May 20, 1983.

DNA topoisomerase activity detected in cell extracts of the trypanosomatid Crithidia fasciculata interlocks kinetoplast DNA duplex minicircles into huge catenane forms resembling the natural kinetoplast DNA networks found in trypanosomes. Catenation of duplex DNA circles is reversible and equilibrium is affected by ionic strength, and by spermidine. The reaction requires magnesium, is ATP dependent and is inhibited by high concentrations of novobiocin. Extensive homology between duplex DNA rings was not required for catenane formation since DNA circles with unrelated sequences could be interlocked into mixed network forms. Covalently sealed catenated DNA circles are specifically used as substrates for decatenation. No such preference for covalently sealed duplex DNA rings was observed for catenane formation. Its catalytic properties and DNA substrate preference, suggest a potential role for this eukaryotic topoisomerase activity in the replication of kinetoplast DNA.


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