Current Opinion in Neurobiology. 231-239 - The neurobiology

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1982) Yeast DNA topoisomerase II. An ATP-dependent type II topoisomerase that catalyzes the catenation, decatenation, unknotting, and relaxation of double-stranded DNA rings. J. Biol. Chem. 257, 5866–5872. 11. Maxwell, A. (1993) The interaction between coumarin drugs and DNA gyrase. Mol. Microbiol. 9, 681–686. 12. Robinson, M. , Corbett, A. , and Osheroff, N. (1993) Effects of topoisomerase II-targeted drugs on enzyme-mediated DNA cleavage and ATP hydrolysis: evidence for distinct drug interaction domains on topoisomerase II.

3. Several classical assays for ATPase activity, including HPLC and TLC are appli- 44 Duguet et al. 2. and labeled ATP (18). 2. 1. Efficient DNA Cleavage Without Drug Since it is a type I topoisomerase, reverse gyrase transiently cleaves a single DNA strand. A cleavage reaction can be observed in the presence of stoichiometric amounts of the enzyme bound to DNA (19,20) (see Note 13). 1. 5 mM EDTA, 30 µg/mL bovine serum albumin, 10 mM MgCl2, 10 mM NaCl, and no ATP. 6 ng of pure enzyme). 2. 5 mg/mL proteinase K.

Reverse Gyrase Activity 49 10. , and, Duguet, M. (1993) ATP-independent DNA topoisomerase from Fervidobacterium islandicum. Biochim. Biophys. Acta 1216, 213–220. 11. Barzilai, R. (1973) SV40 DNA: quantitative conversion of closed circular to open circular form by an ethidium bromide-restricted endonuclease. J. Mol. Biol. 74, 739–742. 12. Nakasu, S. and Kikuchi, A. (1985) Reverse gyrase; ATP-dependent type I topoisomerase from Sulfolobus. EMBO J. 4, 2705–2710. 13. , and Duguet, M. (1988) Reverse gyrase of Sulfolobus: purification to homogeneity and characterization.

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