Analysis of viral and cellular proteins in HIV-1 reverse transcription complexes by co-immunoprecipitation

Abstract

Molecular details and temporal organization of the early (preintegration) phase of HIV life cycle remain among the least investigated and most controversial problems in the biology of HIV. To accomplish reverse transcription and intracellular transport of the viral genetic material, HIV forms multi-molecular complexes termed reverse transcription complexes (RTCs). Analysis of the kinetics of reverse transcription and nuclear import of RTCs, as well as assessment of the changes in their protein content in the course of reverse transcription and nuclear translocation is a necessary step in understanding the mechanisms of cytoplasmic maturation and nuclear import of HIV-1 RTCs. Here, we review methods that allow quantitative assessment of the dynamics of the maturation of HIV-1 RTCs and transformations of RTC protein composition associated with nuclear import of the complexes. © 2009 Humana Press.