Integrase (IN) proteins mediate an essential step in retroviral life cycles, the integration of reverse-transcribed viral DNA into the host genome. To create tools for direct comparative investigations, hexahistidinetagged IN proteins of the phylogenetically related lentivirnses caprine arthritis-encephalitis virus (CAEV), maedi-visna virus (MW) and human immunodeficiency virus type 1 (HIV-1) were expressed in Escherichia coll.After purification by affinity chromatography, the active enzymes were compared in vitro for their site-specific cleavage, integration and disintegration activities on cognate and non-cognate oligonucleotide substrates. It was found that CAEV IN and MW IN catalyse both site-specific cleavage and disintegration with high efficiencies, reduced substrate specificities and similar reaction patterns. Comparisons with the respective activities of HIV-1 IN revealed basic functional similarities as well as considerable differences such as more restricted substrate requirements for site-specific cleavage. On the other hand, all three enzymes catalyse disintegration almost independent of the substrate origin. Furthermore, MVV IN was shown to join oligonucleotides as efficiently as HIV-1 IN, albeit with reduced substrate specificity. In contrast, no detectable strand transfer activities occurred with CAEV IN.

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