FIG 3.

Nup205 associates with HIV Rev via the ARM domain and is required for nuclear localization. (A) H1299 cells were transfected with HA-E4orf4 or HA-Rev or mutants that lack the ARM (ΔARM). HA-SV40-LgT is included as a negative control. Immunoprecipitation from cell extracts was performed using anti-HA. IP complexes were analyzed by Western blotting to detect endogenous Nup205 and HA-tagged viral proteins. Anti-Nup205 Western blotting of whole-cell extract was used to confirm expression of Nup205 (lower portion). (B) Schematic showing the ARMs of E4orf4 and HIV Rev. The nuclear localization sequence of SV40 is shown for comparison. Mutated residues in the ΔARM mutants are indicated in red. (C) Epifluorescence microscopy showing localization of EGFP-fused E4orf4, Rev, and ΔARM mutants.