Nucleoporin NUP153 Phenylalanine-Glycine Motifs Engage a Common Binding Pocket within the HIV-1 Capsid Protein to Mediate Lentiviral Infectivity

• DOI: 10.1371/journal.ppat.1003693
• Publication Date: 2013-10-11T16:24:17Z
• Authors: Kenneth A. Matreyek; Sara S. Yücel; Xiang Li; Alan Engelman
• Keywords: Nucleoporin; Infectivity; HEK 293 cells; Equine infectious anemia; Viral protein

Lentiviruses can infect non-dividing cells, and various cellular transport proteins provide crucial functions for lentiviral nuclear entry integration. We previously showed that the viral capsid (CA) protein mediated dependency on nucleoporin (NUP) 153 during HIV-1 infection, now demonstrate a direct interaction between CA N-terminal domain phenylalanine-glycine (FG)-repeat enriched NUP153 C-terminal (NUP153C). NUP153C fused to effector domains of rhesus Trim5α restriction factor (Trim-NUP153C) potently restricted HIV-1, providing an intracellular readout NUP153C-CA retroviral infection. Primate lentiviruses equine infectious anemia virus (EIAV) bound under these conditions, results correlated with binding purified in vitro. These phenotypes moreover requirement endogenous Mutagenesis experiments concordantly identified residues important infectivity. Different FG motifs within versus EIAV capsids. mapped line common alpha helix 3/4 hydrophobic pocket also mediates small molecule PF-3450074 (PF74) inhibitor cleavage polyadenylation specific 6 (CPSF6) protein, Asn57 (Asp58 EIAV) playing particularly role. PF74 CPSF6 accordingly each competed pocket, significantly higher concentrations were needed inhibit infection face Trim-NUP153C expression or knockdown. Correlation mutant cell cycle dependencies indicates underlies ability cells. Our highlight similar mechanisms disparate host factors same region ingress. conclude subset directly engage FG-motifs present affect import.