A homogeneous type II DNA topoisomerase from HeLa cell nuclei.

K. G. Miller, Leroy-Fong Liu, P. T. Englund

Research output: Contribution to journalArticle

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Abstract

Using kinetoplast DNA networks as a substrate in a decatenation assay, we have purified to apparent homogeneity a type II DNA topoisomerase from HeLa cell nuclei. The most pure preparations contain a single polypeptide of 172,000 daltons as determined by sodium dodecyl sulfate-gel electrophoresis. The molecular weight of the native protein, based on sedimentation and gel filtration analyses, is estimated to be 309,000. These results suggest that the enzyme is a dimer of 172,0090-dalton subunits. The enzyme is a type II topoisomerase as demonstrated by its ability to change the linking number of DNA circles in steps of two and to decatenate or unknot covalently closed DNA circles. No gyrase activity is detectable. ATP is required for the relaxation, decatenation, and unknotting of DNA, and a DNA-dependent ATPase activity is present in the most pure fractions. ATP is hydrolyzed to ADP in this properties to T4 DNA topoisomerase (Liu, L. F., Liu, C. C., and Alberts, B. M. (1979) Nature 281, 456-461).

Original languageEnglish
Pages (from-to)9334-9339
Number of pages6
JournalJournal of Biological Chemistry
Volume256
Issue number17
Publication statusPublished - Sep 10 1981
Externally publishedYes

Fingerprint

Type II DNA Topoisomerase
Cell Nucleus
HeLa Cells
Cells
DNA
Adenosine Triphosphate
Gels
Kinetoplast DNA
Type I DNA Topoisomerase
Enzymes
Electrophoresis
Sedimentation
Sodium Dodecyl Sulfate
Dimers
Adenosine Diphosphate
Gel Chromatography
Adenosine Triphosphatases
Assays
Molecular Weight
Molecular weight

ASJC Scopus subject areas

  • Biochemistry

Cite this

A homogeneous type II DNA topoisomerase from HeLa cell nuclei. / Miller, K. G.; Liu, Leroy-Fong; Englund, P. T.

In: Journal of Biological Chemistry, Vol. 256, No. 17, 10.09.1981, p. 9334-9339.

Research output: Contribution to journalArticle

Miller, KG, Liu, L-F & Englund, PT 1981, 'A homogeneous type II DNA topoisomerase from HeLa cell nuclei.', Journal of Biological Chemistry, vol. 256, no. 17, pp. 9334-9339.
Miller, K. G. ; Liu, Leroy-Fong ; Englund, P. T. / A homogeneous type II DNA topoisomerase from HeLa cell nuclei. In: Journal of Biological Chemistry. 1981 ; Vol. 256, No. 17. pp. 9334-9339.
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