Summary
Investigate HIV-1 reverse transcriptase (RT) structure and function, its roles in the reverse transcription complex (RTC), and viral maturation to advance understanding of HIV replication and guide novel therapeutic strategies.
HIV reverse transcriptase structure, function, inhibition, and roles in viral assembly and maturation
US · IL
NIH
grant
awarded
#nih-2R01AI027690-36A1
What they want
The project proposes to address three critical knowledge gaps: (1) further definition of specific conformational states and transitions during first- and second-strand DNA synthesis initiation and elongation; (2) the molecular interaction between RT and its partner host protein eEF1A and its functional role within the RTC; and (3) the role of the RT portion of Gag-Pol in HIV virion assembly and maturation. This involves determining structures of RT complexes using cutting-edge cryo-EM and X-ray crystallography, complemented by biochemical and virological studies. Efforts also include improving current processing strategies for recovering multiple conformations in cryo-EM data.
Deliverables
- High-resolution structures detailing reverse transcription initiation phases and transition to elongation
- High-resolution structures detailing RT's interactions with eEF1A and rationale for its role in promoting reverse transcription
- High-resolution structures detailing how RT participates in virion assembly within Pol and Gag-Pol polyprotein intermediates
- Insights into how targeted activators of cell kill (TACK) non-nucleoside RT inhibitors enhance PR dimerization and viral maturation, triggering HIV-specific cell death, with potential application to HIV cure strategies
- Identification of new druggable interfaces
- Further characterization of established (non-nucleoside inhibitors) and innovative inhibitor classes (TACK inhibitor molecules)
Technical requirements
- Cryo-EM (cryo-electron microscopy)
- X-ray crystallography
- Biochemical studies
- Virological studies
- Engineering of polyprotein constructs
- Optimization of protein expression and structure determination
- Improvement of current processing strategies for recovering multiple conformations in cryo-EM data
