Dialysis Purification of Integrase-DNA Complexes Provides High-Resolution Atomic Force Microscopy Images: Dimeric Recombinant HIV-1 Integrase Binding and Specific Looping on DNA

Abstract

It remains difficult to obtain high-resolution atomic force microscopy images of HIV-1 integrase bound to DNA in a dimeric or tetrameric fashion. We therefore constructed specific target DNAs to assess HIV-1 integrase binding and purified the complex by dialysis prior to analysis. Our resulting atomic force microscopy analyses indicated precise size of binding human immunodeficiency virus type 1 (HIV-1) recombinant integrase in a tetrameric manner, inducing formation of a loop-like or figure-eight-like secondary structure in the target DNA. Our findings regarding the target DNA secondary structure provide new insights into the intermediate states of retroviral integration. © 2013 Tsuruyama et al.