Rhesus TRIM5α Disrupts the HIV-1 Capsid at the InterHexamer Interfaces
Figure 5
Cross-linked CA assemblies resist structural damage by TRIM5αrh CC-SPRY.
(A) Amino acids at the pseudo three-fold axis in the molecular model of the tubular CA assemblies [38] were used to guide cysteine mutagenesis (P207C/T216C) for cross-linking of CA tubes. (B) Non-reducing SDS-PAGE analysis of TRIM5αrh CC-SPRY (18 µM) binding to cross-linked P207C/T216C CA tubes (left) and cross-linking of P207C/T216C CA tubes after TRIM5αrh CC-SPRY (18 µM) binding (right), visualized by Coomassie Blue staining. Pellets of non-reduced and reduced samples were analyzed in lanes 5&6 and 7&8, respectively. (C&D) CryoEM analysis of the structural effect of TRIM5αrh CC-SPRY binding to P207C/T216C CA tubes without (C, corresponding sample in panel B, lane2) and with cross-linking (D, corresponding sample in panel B, lane4). Some ice particles inadvertently deposited on the EM grid during cryo-sample preparation are visible in panel C (upper right-hand region). The displayed images are representative examples of three independent experiments. Scale bar, 100 nm.