Comparison of enzymatic activities of the HIV-1 and HFV integrases to their U5 LTR substrates.
The human immunodeficiency virus type 1 (HIV-1) and human foamy virus (HFV) integrase proteins were overexpressed in Escherichia coli, and purified to a near homogeneity by one- or two-step purification scheme. The endonucleolytic, integration, and disintegration activities for the HIV-1 and HFV integrases were characterized in vitro. The endonucleolytic activities for the HIV-1 and HFV integrases were found only on their own substrates, respectively, indicating that the cognate U5 LTR sequences in the substrates is critical for specific cleavage. However, the integration and disintegration activities showed less specificity on the substrate usage. Our results suggest that the disintegration activity have more preference for substrates based on Y-shaped structure rather than on viral donor DNA sequence.
D015870:Gene Expression; D019427:HIV Integrase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D019426:Integrases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000235:genetics*; D016092:Spumavirus / Q000201:enzymology*; D013379:Substrate Specificity; D020079:Terminal Repeat Sequences