Isolation and analysis of HIV-1 preintegration complexes.

A discerning feature of the retrovirus lifecycle is the covalent integration of the viral reverse transcript into a chromosome within the infected cell. Integration is required for productive infection and therefore defines the viral integrase protein of human immunodeficiency virus type 1 (HIV-1) as a bona fide target for the development of antiviral drugs in the fight against HIV/AIDS. Integrase works in the context of the viral preintegration complex (PIC), a high molecular weight nucleoprotein complex that supports the integration of its endogenous viral DNA copy made during reverse transcription into an exogenous target DNA in the test tube. PIC analyses are central to understanding the molecular mechanisms of HIV-1 integration as well as investigating the pharmacological properties of integrase inhibitors. This chapter describes techniques for isolating HIV-1 PICs from cells as well as quantifying their level of integration activity in vitro.