Reverse transcription, the process which converts viral genomic RNA into a double strand DNA, is the central defining feature of HIV-1 replication and a major target for anti-retroviral therapy. At the core of the reverse transcription process is a multi-protein complex called the reverse transcription complex composed of reverse transcriptase (RT), viral proteins including IN, CA, and Vpr and cellular proteins including eukaryotic translation elongation factor 1A (eEF1A). We previously showed that eEF1A stabilized the RTC in cells and was important for late steps of reverse transcription. Our experiments show that proteins interacting with the RTC via RT can inhibit reverse transcription by affecting viral uncoating kinetics and RTC stability.
Our recent experiments show a direct interaction between RT and eEF1A that can be down regulated by amino acid substitutions in the RT thumb domain, and which lead to downregulated late reverse transcription. Moreover we show that drugs which bind to eEF1A are potent inhibitors of reverse transcription. Experiments to determine if eEF1A binding drugs negatively affect the RTC in cells will be presented. Recently we also showed that a Tat mutant called Nullbasic inhibits reverse transcription. Our recent data shows that Nullbasic is an RT binding protein that is found in viral particles. In vitro uncoating assays show that virions containing Nullbasic undergo accelerated uncoating kinetics, and analysis of cells infected with HIV-1 containing Nullbasic indicates that the levels of RTCs are reduced; consistent with the uncoating defect. The combined evidence indicates investigations of interaction between RT and viral and cellular proteins could enable new antiviral strategies.