Figure 6
From: Highly conserved serine residue 40 in HIV-1 p6 regulates capsid processing and virus core assembly
S40F mutation has no effect on ALIX mediated virus release. (A) Rescue of budding of the Ser-40 mutant by ALIX. Virus release, Gag processing, and exogenous expression of ALIX were analyzed by Western blot (upper panel). Ribosomal P0 antigen (RP0) was used as loading control. The amounts of infectious units released from the cells were analyzed by β-galactosidase quantification after infection of TZM-bl cells (lower panel). Shown are virus infectivities relative to the HIV-1ΔPTAP control ± SD. Lane 1 shows HIV-1ΔPTAP mutant cotransfected with the empty control vector, lane 2 shows HIV-1ΔPTAP mutant cotransfected with a FLAG-ALIX expression plasmid, lanes 3 and 4 show the HIV-1ΔPTAP/ΔYP double mutant cotransfected with the empty control plasmid and the vector expressing V5-POSH, respectively, lanes 5 and 6 show the HIV-1ΔPTAP/ΔYP double mutant cotransfected with the empty control plasmid and the vector expressing V5-POSH, respectively. (B) Overexpression of ALIX does not rescue infectivity of the S40F mutant. The amounts of infectious units released from the cells analyzed were by β-galactosidase quantification after infection of TZM-bl cells. Shown are virus infectivities relative to the HIV-1 control ± SD. 1: HIV-1 contransfected with the empty control vector, 2: HIV-1 cotransfected with a FLAG-ALIX expression plasmid, 3: HIV-1S40F cotransfected with the empty control vector, 2: HIV-1S40F cotransfected with a FLAG-ALIX expression plasmid.