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The EMBO Journal (2004) 23, 897–907, doi:10.1038/sj.emboj.7600077 Published online 5 February 2004 Figure 2 DNA topology, not DNA sequence, is a critical determinant for Drosophila ORC–DNA binding Dirk Remus, Eileen L Beall and Michael R Botchan

Figure 2 rDmORC binds with similar affinity to fragments of the third chromosome chorion gene cluster. ATP-dependent binding of rDmORC to a radiolabeled ACE-3 fragment was assayed by EMSA in the presence of competitor fragment DNA indicated in the upper right corner of each plot. % binding is relative to the fraction of ACE-3 bound in the absence of competitor. (A) Relative binding efficiencies of rDmORC (left two panels) and embryonic (0–12 h) DmORC to ACE-3 in the presence of increasing amounts of ACE-3 and ACE-D competitor. Dashed lines indicate [C]1/2. (B) Binding plots of all fragments tested. The relative position of each fragment in the chorion gene cluster is schematized in the center. (C) Summary plot depicting [C]1/2 for each fragment normalized to ACE-3. The average and standard deviation for two to three independent experiments are shown.

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