FIGURE 8.

ICP27 blocks both NLS and M9-mediated nucleocytoplasmic transport pathways. A, left panels, ICP27 accumulates both in the nucleoplasm and the cytoplasm of uninfected cells transfected with a WT ICP27-encoding plasmid. Middle panels, HeLa cells were transfected with a plasmid encoding just a NES-GFP₂-M9 reporter that contains the CRM1-dependent NES of HIV-1 Rev and the transportin-dependent M9-NLS of hnRNPA1. This construct accumulates in the nucleoplasm when transfected alone but in the cytoplasm when co-transfected with ICP27 (right panels). B, NES-GFP₂-M9 reporter construct when co-transfected with ICP27 encoded from pTriEx-ICP27 WT plasmid also accumulates predominantly in the cytoplasm. C, similarly, a different reporter was used (NES-GFP₂-cNLS) that contains the CRM1-dependent REV NES and the importin/karyopherin α/β-dependent (classical) NLS. It accumulates in the nucleoplasm when expressed alone (left panels) and in the cytoplasm when both ICP27 and NES-GFP₂-cNLS are co-transfected (right panels). Scale bar, 10 μm.