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PMID: 10590138 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

cis expression of the F12 human immunodeficiency virus (HIV) Nef allele transforms the highly productive NL4-3 HIV type 1 to a replication-defective strain: involvement of both Env gp41 and CD4 intracytoplasmic tails.

Journal of virology ·Vol. 74 ·No. 1 ·2000-01-00 ·Pages 483-92

Olivetta E, Pugliese K, Bona R, D'Aloja P, Ferrantelli F, Santarcangelo AC, Mattia G, Verani P, Federico M

Abstract

F12 human immunodeficiency virus type 1 (HIV-1) nef is a naturally occurring nef mutant cloned from the provirus of a nonproductive, nondefective, and interfering HIV-1 variant (F12-HIV). We have already shown that cells stably transfected with a vector expressing the F12-HIV nef allele do not downregulate CD4 receptors and, more peculiarly, become resistant to the replication of wild type (wt) HIV. In order to investigate the mechanism of action of such an HIV inhibition, the F12-HIV nef gene was expressed in the context of the NL4-3 HIV-1 infectious molecular clone by replacing the wt nef gene (NL4-3/chi). Through this experimental approach we established the following. First, NL4-3/chi and nef-defective (Deltanef) NL4-3 viral particles behave very similarly in terms of viral entry and HIV protein production during the first replicative cycle. Second, no viral particles were produced from cells infected with NL4-3/chi virions, whatever the multiplicity of infection used. The viral inhibition apparently occurs at level of viral assembling and/or release. Third, this block could not be relieved by in-trans expression of wt nef. Finally, NL4-3/chi reverts to a producer HIV strain when F12-HIV Nef is deprived of its myristoyl residue. Through a CD4 downregulation competition assay, we demonstrated that F12-HIV Nef protein potently inhibits the CD4 downregulation induced by wt Nef. Moreover, we observed a redistribution of CD4 receptors at the cell margin induced by F12-HIV Nef. These observations strongly suggest that F12-HIV Nef maintains the ability to interact with the intracytoplasmic tail of the CD4 receptor molecule. Remarkably, we distinguished the intracytoplasmic tails of Env gp41 and CD4 as, respectively, viral and cellular targets of the F12-HIV Nef-induced viral retention. For the first time, the inhibition of the viral life cycle by means of in-cis expression of a Nef mutant is here reported. Delineation of the F12-HIV Nef mechanism of action may offer additional approaches to interference with the propagation of HIV infection.

MeSH Terms
Alleles CD4 Antigens/physiology Cell Line Defective Viruses/genetics,physiology Down-Regulation Gene Expression Regulation, Viral Genes, nef HIV Envelope Protein gp41/physiology HIV-1/genetics,physiology Humans Membrane Fusion Virion
Chemicals
CD4 Antigens HIV Envelope Protein gp41
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Olivetta E
Laboratory of Virology, Istituto Superiore di Sanità, Rome, Italy.
Pugliese K
Bona R
D'Aloja P
Ferrantelli F
Santarcangelo A C
Mattia G
Verani P
Federico M
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.1.483-492.2000
Published
2000-01-00
Pages
483-92
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC111560
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.74.1.483-492.2000 PMC Full Text View journal details
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