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DNA supercoiling by DNA gyrase

A static head analysis

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Abstract

Using purified DNA gyrase to supercoil circular plasmid pBR322 DNA, we examined how the linking number attained at the steady state (‘static head’) varies with the concentrations of ATP and ADP, both in the absence and presence of spermidine. In the absence of spermidine at total adenine nucleotide concentrations between 0.35 and 1.4 mM, the static-head linking number was independent of the sum concentration of ATP and ADP, but depended strongly on the ratio of their concentrations. We established that the same linking number was attained independent of the direction from which the steady state was approached. The decrease in linking number at static head is more extensive when spermidine is present in the incubation, but remains a function of the [ATP]-to-[ADP] ratio.

These results are discussed in terms of various kinetic schemes for DNA gyrase. We present one kinetic scheme that accounts for the experimental observations. According to this scheme our experimental results imply that there is significant slip in DNA gyrase when spermidine is absent. It is possible that spermidine acts through adjustment of the degree of coupling of DNA gyrase.

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Authors and Affiliations

  1. Laboratory of Molecular Biology, National Institute of Diabetes, and Digestive and Kidney Diseases, National Institutes of Health, 20892, Bethesda, MD

    Hans V. Westerhoff, Mary H. O’Dea & Martin Gellert

  2. Department of Biochemistry, University of Leicester, LE1 7RH, Leicester, UK

    Anthony Maxwell

Authors
  1. Hans V. Westerhoff
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  2. Mary H. O’Dea
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  3. Anthony Maxwell
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  4. Martin Gellert
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Additional information

Parts of this study have been presented earlier in a preliminary form (1, 2).

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Westerhoff, H.V., O’Dea, M.H., Maxwell, A. et al. DNA supercoiling by DNA gyrase. Cell Biophysics 12, 157–181 (1988). https://doi.org/10.1007/BF02918357

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  • DOI: https://doi.org/10.1007/BF02918357

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