Immature HIV-1 assembles from Gag dimers leaving partial hexamers at lattice edges as substrates for proteolytic maturation.

Tan, A.; Pak, A. J.; Morado, D. R.; Voth, G. A.; Briggs, J. A. G.

doi:10.1101/2020.10.01.322081

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bioRxiv · 10.1101/2020.10.01.322081

Immature HIV-1 assembles from Gag dimers leaving partial hexamers at lattice edges as substrates for proteolytic maturation.

Tan, A.·Pak, A. J.·Morado, D. R.·Voth, G. A.·Briggs, J. A. G.

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Abstract

The CA (capsid) domain of immature HIV-1 Gag and the adjacent spacer peptide 1 (SP1) play a key role in viral assembly by forming a lattice of CA hexamers, which adapts to viral envelope curvature by incorporating small lattice defects and a large gap at the site of budding. This lattice is stabilized by intra- and inter-hexameric CA-CA interactions, which are important in regulating viral assembly and maturation. We applied subtomogram averaging and classification to determine the structure of CA at lattice edges and found that they form partial hexamers. These structures reveal the network of interactions formed by CA-SP1at the lattice edge. We also performed atomistic molecular dynamics simulations of CA-CA interactions stabilizing the immature lattice and of partial CA-SP1 helical bundles. Free energy calculations reveal increased propensity for helix-to-coil transitions in partial hexamers compared to complete six-helix bundles. Taken together, these results suggest that the CA dimer is the basic unit of lattice assembly, that partial hexamers exist at lattice edges, that these are in a helix-coil dynamic equilibrium and that partial helical bundles are more likely to unfold, representing potential sites for HIV-1 maturation initiation. Significance StatementHIV-1 particle assembly is driven by the viral Gag protein, which oligomerizes into an hexameric array on the inner surface of the viral envelope, forming a truncated spherical lattice containing large and small gaps. Gag is then cut by the viral protease, disassembles and rearranges to form the mature, infectious virus. Here, we present structures and molecular dynamics simulations of the edges of the immature Gag lattice. Our analysis shows that Gag dimers are the basic assembly unit of the HIV-1 particle, that lattice edges are partial hexamers, and that partial hexamers are prone to structural changes allowing protease to cut Gag. These findings provide insights into assembly of the immature virus, its structure, and how it disassembles during maturation.

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Tan, A., Pak, A. J., Morado, D. R., Voth, G. A., Briggs, J. A. G.. 2020-10-01. Immature HIV-1 assembles from Gag dimers leaving partial hexamers at lattice edges as substrates for proteolytic maturation.. https://doi.org/10.1101/2020.10.01.322081

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