Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2000 May;74(10):4891-3.
doi: 10.1128/jvi.74.10.4891-4893.2000.

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

H Akari  1 , T Fukumori, A Adachi
Affiliations

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.

PubMed Disclaimer

Figures

FIG. 1
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
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
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
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).
See this image and copyright information in PMC

References

    1. Adachi A, Gendelman H E, Koenig S, Folks T, Willey R, Rabson A, Martin M A. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone. J Virol. 1986;59:284–291. - PMC - PubMed
    1. Adachi A, Ono N, Sakai H, Ogawa K, Shibata R, Kiyomasu T, Masuike H, Ueda S. Generation and characterization of the human immunodeficiency virus type 1 mutants. Arch Virol. 1991;117:45–58. - PubMed
    1. Akari H, Uchiyama T, Fukumori T, Iida S, Koyama A H, Adachi A. Pseudotyping HIV-1 by vesicular stomatitis virus G protein does not reduce the cell-dependent requirement of Vif for optimal infectivity: functional difference between Vif and Nef. J Gen Virol. 1999;80:2945–2949. - PubMed
    1. Boge M, Wyss S, Bonifacino J S, Thali M. A membrane proximal tyrosine-based signal mediated internalization of the HIV-1 envelope glycoprotein via interaction with the AP-2 clathrin adaptor. J Biol Chem. 1998;273:15773–15778. - PubMed
    1. Dubay J W, Roberts S J, Hahn B H, Hunter E. Truncation of the human immunodeficiency virus type 1 transmembrane glycoprotein cytoplasmic domain blocks virus infectivity. J Virol. 1992;66:6616–6625. - PMC - PubMed

Publication types

Substances

LinkOut - more resources