FIGURE 4.

MCV LT.W209A mutant is defective in interacting with and relocalizing hVam6p. A, mutation at Trp-209 on full-length LT completely abolished LT-hVam6p binding. Lysates from U2OS cells transiently co-expressing HA-hVam6p with empty vector, LT-V5, or LT.W209A-V5 were immunoprecipitated (IP) for LT with anti-V5 antibody and immunoblotted (IB) for hVam6p by anti-HA antibody. B, LT.W209A fails to relocalize hVam6p to the nucleus. U2OS cells were co-transfected with Myc-hVam6p and LT-DsRed or LT.W209A-DsRed. hVam6p is visualized with anti-Myc antibody and Alexa Fluor 488-conjugated anti-mouse antibody (FITC), whereas the wild-type and mutant LTs are visualized by DsRed fluorescence.