FIG. 5.

Effect of a primary or combined primary and late cleavage site mutations within Gag or Gag-Pol on intracellular viral proteins and virion protein profiles of HIV-1. Intracellular viral proteins were standardized by EGFP, and total virion protein levels were standardized by dot blot analysis (data not shown). Cotransfected cell lysates (A) and purified virions (see Materials and Methods) (B) were resolved by SDS–10% PAGE. Resolved proteins were probed using sera from HIV-1-infected individuals, as described in Materials and Methods. Lanes 1, show G/GP cellular viral proteins and virion protein profiles, respectively. Lanes 2 and lanes 3 show cellular viral proteins and virion protein profiles of mutant viruses containing a primary CA2 (p2/NC) or a combined primary and late cleavage site mutation CA6 (p2/NC, CA/p2, and a cryptic site mutation) in Gag-Pol, respectively. Lanes 4 and lanes 5 show cellular viral proteins and virion protein profiles of mutant viruses containing a primary CA2 (p2/NC) or a combined primary and late cleavage site mutation CA6 (p2/NC and CA/p2) in Gag, respectively. Lanes a and lanes b show mock- and WT-transfected controls. The amounts of WT HIV-1 intracellular and virion proteins were independent of the other proteins tested and were used in these analysis only to compare HIV-1 protein patterns probed by HIV-1 sera.