AIDS Innovation Project

National Institute Of Arthritis And Musculoskeletal And Skin Diseases

Investigators

Abstract

We have determined the structures of Rev–RRE complexes across various native RRE lengths and conformations using cryo-EM. Determined structures include reconstituted complexes of Rev with wild-type short (~234nt) and long (~332nt) RREs, and sequence variants favoring 4SL versus 5SL architectures. The structures reveal a Rev-mediated bridge between stem IIB and stem IA, demonstrating multi-site engagement across spatially separated RNA elements. The results offers a concrete architectural rationale for cooperativity and long-range coupling within the RRE; pinpoints structural “hotspots” for variant mapping and functional tests. We have used longitudinal patient sequence data to identify natural Rev and RRE variants. Future work will solve or model these variants using cryo-EM data and map onto existing Rev–RRE structures. Variants are also being mapped onto solved structures to generate interface-centric heat maps and to prioritize patient- relevant constructs for targeted reconstructions. The successful acquisition and structural analysis of RRE constructs lay a foundation for understanding the molecular consequences of clinically relevant mutations. These findings offer new insights into the diversity of Rev-RRE interactions and provide a structural basis for the design of future therapeutic interventions targeting HIV-1 RNA export.