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HIV-1 Packaging Visualised by In-Gel SHAPE.

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D'Souza, Aaron R 
Jayaraman, Dhivya 
Prestwood, Liam J 


HIV-1 packages two copies of its gRNA into virions via an interaction with the viral structural protein Gag. Both copies and their native RNA structure are essential for virion infectivity. The precise stepwise nature of the packaging process has not been resolved. This is largely due to a prior lack of structural techniques that follow RNA structural changes within an RNA-protein complex. Here, we apply the in-gel SHAPE (selective 2'OH acylation analysed by primer extension) technique to study the initiation of HIV-1 packaging, examining the interaction between the packaging signal RNA and the Gag polyprotein, and compare it with that of the NC domain of Gag alone. Our results imply interactions between Gag and monomeric packaging signal RNA in switching the RNA conformation into a dimerisation-competent structure, and show that the Gag-dimer complex then continues to stabilise. These data provide a novel insight into how HIV-1 regulates the translation and packaging of its genome.



HIV-1, packaging, in-gel SHAPE, Gag, NC, RNA structure, dimerisation

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Medical Research Council (MR/N022939/1)
This work was funded by an Eric Reid fund for Methodology grant from the Biochemical Society (to JCK), UK Medical Research Council, Grant/Award Number: MR/N022939/1 (to AML and JCK). AMLL is supported by the Clinical Academic Reserve and his laboratory by the NIHR Cambridge BRC (Grant RCAG/18). Research in the Kappei laboratory was supported by the National Research Foundation Singapore and the Singapore Ministry of Education under its Re-search Centres of Excellence initiative.