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The EMBO Journal (2004) 23, 897–907, doi:10.1038/sj.emboj.7600077 Published online 5 February 2004 Figure 4 DNA topology, not DNA sequence, is a critical determinant for Drosophila ORC–DNA binding Dirk Remus, Eileen L Beall and Michael R Botchan

Figure 4 DmORC binds preferentially to (-) supercoiled DNA. rDmORC binding to 125 fmol 3H-labeled pBS/ori- was assayed by nitrocellulose filter binding. (A) Titration of rDmORC with 0.5 mM ATP. (B) Titration of ATP with 1 pmol rDmORC. (C) rDmORC binding to 3H-labeled (-) supercoiled pBS/ori- in the presence of increasing amounts of unlabeled (-) supercoiled pBS/ori- (), pBS/ori- relaxed with TopoI (), pBS/ori- linearized with BamHI (), or (-) supercoiled pBluescript (). Inset panel: Ethidium-bromide-stained agarose gel showing (-) supercoiled, linearized, and relaxed pBS/ori-.

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