Cell-dependent requirement of human immunodeficiency virus type 1 gp41 cytoplasmic tail for Env incorporation into virions - PubMed (original) (raw)

Cell-dependent requirement of human immunodeficiency virus type 1 gp41 cytoplasmic tail for Env incorporation into virions

H Akari et al. J Virol. 2000 May.

Abstract

Growth kinetics in lymphocytic H9 and M8166 cells of two mutants of human immunodeficiency virus type 1 (HIV-1) with deleted gp41 cytoplasmic tails were examined. While the mutant viruses designated CTdel-44 and CTdel-144 were able to grow in M8166 cells, they were unable to grow in H9 cells. Transfection and single-round infectivity assays demonstrated that they are defective in the early phase of viral replication in H9 cells. Analysis of the mutant virions revealed drastically reduced incorporation of Env gp120 (compared with the incorporation of wild-type virions) in H9 cells but normal incorporation in M8166 cells. These results indicate that the HIV-1 cytoplasmic tail of gp41 determines virus infectivity in a cell-dependent manner by affecting incorporation of Env into virions and suggest the involvement of a host cell factor(s) in the Env incorporation.

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Figures

FIG. 1

Growth kinetics of wt and mutant viruses in H9 and M8166 cells. Viruses were prepared from HeLa cells transfected with pNL432 (WT), pNL-Nd (vif mutant [Nd]), CTdel-44, or CTdel-144. Cells (106) were infected with 5 × 105 RT units of viruses. RT production in the culture supernatants was monitored at intervals.

FIG. 2

Infectivity of wt and mutant viruses. Single-round infectivity of viruses derived from H9 and M8166 cells electroporated with pNL432 (WT), pNL-Nd (vif mutant [Nd]), pNL-Kp (env mutant [Kp]), CTdel-44 (44), or CTdel-144 (144) was monitored by MAGI assay. Infectivity was determined by counting blue foci of X-Gal-treated MAGI cells 2 days after inoculation of viruses. Average and standard deviation of three independent experiments are shown.

FIG. 3

Analysis of virion proteins by Western blotting. The lysates of virions prepared from H9 (A to C) and M8166 (D to F) cells electroporated with pNL432 (WT), pNL-Nd (vif mutant [Nd]), CTdel-44, CTdel-144, or pUC19 (control) were analyzed by Western blotting with a sheep anti-Env gp160 antiserum (A and D), a mouse anti-p24 monoclonal antibody (B and E), and a sheep anti-RT antiserum (C and F).

FIG. 4

Analysis of viral proteins expressed in cells by Western blotting. The lysates of H9 (A and B) and M8166 (C and D) cells electroporated with pNL432 (WT), pNL-Nd (vif mutant [Nd]), CTdel-44, CTdel-144, or pUC19 (control) were analyzed by Western blotting with a sheep anti-Env gp160 antiserum (A and C) and a mouse anti-p24 monoclonal antibody (B and D).

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Cell-dependent requirement of human immunodeficiency virus type 1 gp41 cytoplasmic tail for Env incorporation into virions - PubMed (original) (raw)