Figure 1

Inhibition of HIV replication by drugs that block eIF5A modification. A. Hypusination of eIF5A (gray) occurs in two steps: the transfer, catalyzed by DHS, of an aminobutyl moiety (blue) from spermidine onto the side chain of eIF5A lysine-50, yielding deoxyhypusine (Dhp); and its subsequent hydroxylation, catalyzed by DOHH, yielding hypusine (Hpu). DHS is inhibited by GC7 and DOHH by CPX and DEF, as indicated. B. Structures of CPX, Agent P2, DEF and DFOX. C. CPX and DEF inhibit HIV replication in infected PBMCs. Infected PBMCs that were isolated from a single donor were co-cultured with uninfected PBMCs. CPX (30 μM), P2 (30 μM), or DEF (250 μM) were added 48 hr later. Amount of released p24 protein per million viable cells was determined every 24 hr. D. CPX and DEF inhibit gene expression from an HIV molecular clone in a dose dependant manner. The molecular clone pNL4-3-LucE^- and pCMV-Ren were transfected into 293T cells and drugs were added to the concentrations shown. Dual luciferase assays were conducted at 12 hr post-transfection. Firefly (FF) luciferase expression was normalized to Renilla luciferase (Ren) from pCMV-Ren (mean of 2 experiments in duplicate, ± SD). Inset shows CPX and DEF effects on apoptosis and cell viability in untransfected 293T cultures as measured by staining with annexin V (AnnV) and 7-amino-actinomycin D (7AAD). Data are means of three time points (12, 18 and 24 hr) presented as percentages.