Figure 5.

Role of NP late-domain motifs in the enhancement of TCRV Z-induced budding. (A) TCRV NP and Z both interact with the ESCRT-binding component Alix/AIP1. Co-immunoprecipitation studies using HA-agarose were performed 48 h after transfection of 293T cells with pCAGGS Alix-flag alone or in combination with pCAGGS TCRV NP-HA or TCRV Z-HA. (B) Schematic representation of TCRV NP and its putative late-domains. Mutants in which individual late domains are knocked-out (NP_1AxxA, NP_2AxxA, NP_θAxxA) by exchanges to alanine at the indicated positions are also shown. (C) TCRV Z-directed budding and the incorporation of NP late-domain mutants. VLPs were generated as previously described [80]. The release of Z in combination with NP was set at 100% and the relative budding efficiencies of Z alone or together with the various NP mutants shown in (B) were calculated. Release of Z in VLPs is shown in black while release of NP is shown in grey. Data represent the mean value and standard deviation of three independent experiments. The statistical significance was determined using Student’s t test. Asterisks indicate statistically significant differences (*p<0.05, **p<0.01, ***p<0.001).