Journal: Cell

Article Title: Cryo-ET of Env on intact HIV virions reveals structural variation and positioning on the Gag lattice

doi: 10.1016/j.cell.2022.01.013

Figure Lengend Snippet: Structural analyses of Env sub-tomogram averaged maps from mature and immature hVLPs, related to , , , and (A) Perpendicular slices through the C3-symmetrized hVLP-Env map. (B) Gold-standard Fourier shell correlation (FSC) curves for C3-symmetrized map (blue line) and C1-symmetry map (gray line). Dotted line indicated the 0.143 FSC cutoff. (C–F) Local resolution estimation using Resmap software ( Kucukelbir et al., 2014 ) for C3-symmetrized map (C and D) and asymmetric map (E and F). (G) Fourier shell correlation (FSC) curves for C3 (gray line) and C1 (blue line) symmetrized maps of immature hVLP-Env. (H) Top and side surface views of immature hVLP-Env structure with C1 symmetry, C3 symmetry and the C3 map relaxed to C1 symmetry. (I) Top view of Gag layer from corresponding averages in panel B. (J) Left: Previously published high resolution density map of Gag-CA lattice, low pass filtered to ∼10 Å ( Mattei et al., 2018 ). Right: Gag-CA structure derived after focused refinement of Gag layer from the relaxed C1 symmetry immature Env map. (K) HIV protease inhibitor darunavir blocks the processing of immature pr55 Gag precursor into the mature p24 capsid protein. ADA.CM Env with 102 residues truncation in CTD from hVLPs and full-length Env-CTD from pseudovirus, produced in the presence (+) or absence (-) of darunavir, were separated by SDS-PAGE and probed by western blot staining with an anti-p24 antibody.

Article Snippet: For the non-AT-2 treated immature ADA.CM.v4 hVLPs and immature full-length ADA.CM pseudovirus, frozen grids were imaged on a 300 kV Titan Krios equipped with a Gatan K3 direct electron detector and a GIF energy filter with a slit width of 20 eV.

Techniques: Software, Derivative Assay, Protease Inhibitor, Produced, SDS Page, Western Blot, Staining