HIV-1 capsid stability enables inositol phosphate-independent infection of target cells and promotes integration into genes
Fig 3
CA T200I suppresses IP5/6 dependence by HIV-1 CA mutants.
(A) Passaging of HIV-1R9 CA Q219A to select for adaptive mutations. Virus spread was monitored by exogenous RTN using the cell-free culture medium. Virus from the HIV-1 CA Q219A 25 ng p24 infection at ~ 36 days post infection was used to infect CEM cells to create passage 2 (P2) spread assays. P2-A and P2-B are independent P2 spread assays, in P2-B the virus was spin inoculated onto the cells. Passage 3-A (P3) and P3-B spread assays were inoculations of P2-A and P2-B, respectively, onto new CEM cells. Asterisks indicate that the indicated genotype was not the primary HIV-1 variant by Sanger sequencing. For genes with multiple genotypes (i.e. CA), mutations lacking an asterisk indicate the primary genotype. (B and C) Target cell infection of CEM WT cells (B) or the indicated cell lines (C) with the indicated HIV-1GFP reporter virus as determined by flow cytometry. (D) Relative amounts of early (FST) and late (full length minus (FLM)) RTN products. For asterisk(s) without comparison brackets, statistical significance within a given CA mutant is color coded and compares IPMK KOVector to the corresponding (color-coded) control cell line. In (D), HIV-1 CA T200I statistics compare early and late products produced in the same cell line. All bars represent the average of 3 or more independent experiments. Significance levels: p < 0.05 *, p < 0.01 **, p < 0.001 ***, and p < 0.0001 ****.