Stability and conformation of the dimeric HIV-1 genomic RNA 5’UTR

Abstract During HIV-1 assembly the viral Gag polyprotein specifically selects the dimeric RNA genome for packaging into new virions. The 5' untranslated region (5'UTR) of the dimeric genome may adopt a conformation that is optimal for recognition by Gag. Further conformational rearrangement of the 5'UTR, promoted by the NC domain of Gag, is predicted during virus maturation. Two 5'UTR dimer conformations, the kissing dimer and the extended dimer, have been identified in vitro, which differ in the extent of intermolecular base pairing. Whether 5'UTRs from different HIV-1 strains with distinct sequences have access to the same dimer conformations has not been determined. Here, we applied fluorescence cross-correlation spectroscopy (FCCS) and single-molecule Forster resonance energy transfer (smFRET) imaging to demonstrate that 5'UTRs from two different HIV-1 subtypes form kissing dimers with divergent stabilities. We further show that both 5'UTRs convert to a stable dimer in the presence of the viral NC protein, adopting a conformation consistent with extensive intermolecular contacts. These results support a unified model in which the genomes of diverse HIV-1 strains adopt an extended dimer conformation.