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J. Biochem, 1982, Vol. 91, No. 4 1337-1342
© 1982 Japanese Biochemical Society

research-article

Isolation and Partial Characterization of Two Distinct DNA Topoisomerases from Cauliflower Inflorescence1

Hideki FUKATA and Hirosuke FUKASAWA

Department of Biology, Faculty of Science, Kobe University Kobe, Hyogo 657

DNA topoisomerases which remove superhelical turns in closed circular DNA have been isolated from cauliflower inflorescences using polyethylene glycol fractiona-tion, ammonium sulfate precipitation, and column chromatography on CM-Se-phadex or CM-cellulose and DNA-cellulose. Two distinct enzymes, topoisomerase-I and ATP-dependent topoisomerase, were separated clearly by CM-Sephadex or CM-cellulose, and partially characterized using agarose gel electrophoresis with plasmid pBR322 DNA. Topoisomerase-I acts like other eucaryotic DNA topoisomerases in the absence of ATP, is stimulated by spermidine and inhibited by EDTA. The ATP-dependent topoisomerase acts like topoisomerase-I only in the presence of ATP in the reaction medium, is inhibited by spermidine and EDTA, and does not introduce supertwists into closed duplex DNA or produce catenate aggregates under the present reaction conditions.

1This study was supported in part by a grant from the Ministry of Education, Science and Culture of Japan.


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