Nucleoporin NUP153 Phenylalanine-Glycine Motifs Engage a Common Binding Pocket within the HIV-1 Capsid Protein to Mediate Lentiviral Infectivity (original) (raw)
Figure 1
NUP153C directly binds the HIV-1 CA N-terminal domain.
(A) Schematic of NUP153 protein, with residue numbers of domain boundaries indicated. (B) Full length or truncated fragments of HA-tagged NUP153 extracted from 293T cells were tested for binding to HIV-1 CA-NC. Pelleted proteins were resolved by SDS-PAGE and visualized by western blotting with anti-HA antibody 3F10 (top), or by Coomassie stain (bottom). Input, 20% of binding reaction. CA-NC was included in the binding reactions as indicated. (C) Recombinant, tag-free NUP153C and GST purified from E. coli were similarly tested for binding to CA-NC; proteins were detected with Coomassie stain. (D) Recombinant NUP153C pulled down with full length his-tagged wild-type (WT) or W184A/M185A HIV-1 CA, and detected with Coomassie stain. (E) Recombinant NUP153C pulled down with his-tagged CAN, and detected with Coomassie stain. Each experiment was repeated at least 3 times, with a single representative result shown.