As the major target for therapy, Reverse Transcriptase (RT), and in particular the RT from the HIV virus, has been the subject of extensive research. However, despite an ever-increasing amount of information from traditional “bulk” studies, the detailed mechanisms of polymerization are still not completely understood. This is due, partially, to the stochastic nature of individual enzymes, which makes it impossible to follow more than one or two individual steps of polymerization using ensemble methods. I propose here to conduct a single-molecule, mechanistic study of HIV RT, by developing a high-resolution optical-tweezers, capable of resolving the individual polymerization and translocation steps of RT at a broad range of forces and nucleotides concentrations. Characterizing the dwell-time distribution at different experimental condition will allow us to study the complete mechano-chemical cycle of RT, its regulation by the template’s backbone-composition, structure and sequence, its interaction with additional viral and host factors, the molecular basis of inhibition by known inhibitors and the mechanism of resistance acquired through different mutations.
Field of science
- /medical and health sciences/health sciences/infectious disease/RNA virus/hiv
Call for proposal
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