FIG. 6.

CA-G225S compensates for the replication defect imposed by SP1-A3V. (A) Jurkat T cells were transfected with the indicated WT or mutant molecular clones. Cultures were maintained either without or with 1.0 μg/ml PA-457. Cells were split every 2 days, and supernatants were reserved at each time point for RT analysis. (B and C) Effect of the CA-G225S mutation on the extent of cleavage of CA-SP1 to CA. HeLa cells were transfected with WT pNL4-3 or derivatives containing the indicated mutations and pulse-labeled for 15 min with [³⁵S]Met/Cys. Cells were chased for the indicated times, cell lysates were immunoprecipitated with HIV-Ig, and the extent of processing of CA-SP1 to CA was analyzed by SDS-PAGE and fluorography (B) followed by phosphorimager analysis to quantify the percentage of CA-SP1 relative to total CA-SP1 plus CA (C). Error bars indicate standard errors of the means (n = 3).