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. Author manuscript; available in PMC: 2011 Jun 20.
Published in final edited form as: Virology. 2010 Apr 7;402(1):94–101. doi: 10.1016/j.virol.2010.03.001

Figure 2. Enhanced binding of Rep78 to multiple AAV DNA substrates in the presence of E1.

Figure 2

32P-labeled DNA substrates, as indicated, were incubated with the indicated amounts of MBP-Rep78 in the absence or presence of GST-E1 and analyzed by EMSA. Numbers indicate the amount of the added proteins in micrograms. The positions of free probe and probe bound in a Rep complex are indicted as “free” and “bound”. (A) Binding of MBP-Rep78 to complete hairpin ITR DNA was enhanced by GST-E1 in a dose-dependent manner. (B) Binding of MBP-Rep78 to ITR-stem was also enhanced by GST-E1 in a dose-dependent manner. (C) Binding of MBP-Rep78 to p5 promoter DNA was also enhanced by GST-E1 in a dose-dependent manner. (D) Survival of the MBP-Rep78-ITR complex over time was enhanced by GST-E1. (E) Graphical representation of enhanced stability of Rep78-ITR complex in presence of E1 after challenging preformed complex with unlabeled ITR substrate for 20 min.