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. 2008 Sep 12;36(18):5822–5831. doi: 10.1093/nar/gkn560

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Metnase promotes Topo-IIα-mediated decatenation of kDNA. (A) Purified recombinant Metnase nicks supercoiled plasmid to relaxed, open circular and linearized plasmid but does not degrade it. (B) Purified Metnase decatenates kDNA, forming linear DNA and a smaller amount of nicked, open circular DNA. M1, M2-markers for kDNA forms as indicated. (C) Purified Metnase enhances Topo IIα decatenation of kDNA, forming relaxed closed circular DNA. At concentrations of Metnase and Topo IIα where neither alone has an appreciable decatenation activity, they act synergistically to decatenate kDNA to nicked open circular and relaxed closed circular DNA. (D) Graph of dose–response of the enhancement of Topo IIα decatenation by Metnase. Densitometric analysis of open circular and closed circular versus well kDNA for relative kDNA decatenation from n = 3, SD too small for symbols. (E) ICRF-193 does not abrogate Metnase enhancement of Topo IIα decatenation. (F) Graph of densitometric scans of kDNA decatenation performed in the presence of ICRF-193 shown in E, n = 3, SD too small for symbols. (G) Metnase lacking nuclease activity [D483A, ref. 9)] loses most but not all of its ability to enhance Topo IIα decatenation. (H) Graph of densitometry from three D483A experiments, SD shown where larger than symbol. (I) DNase I nicks, linearizes and ultimately degrades kDNA, as opposed to Metnase activity in B.