Association of p56lck with the cytoplasmic domain of CD4 modulates HIV-1 expression.
To investigate the role played by the cytoplasmic domain of the CD4 glycoprotein in the process of HIV infection, we have transfected two CD4-negative human T cell lines with cDNAs encoding the full-length CD4 and a truncated form of the molecule, lacking most of the cytoplasmic domain. Levels of viral replication were significantly higher in cells carrying the truncated version of CD4, in comparison with cells expressing the full-length CD4, as measured by the percentage of cells expressing viral p24 protein and the number of infectious particles released into culture supernatants. The extent of viral entry and reverse transcription was similar in each case, as monitored by an enzymatic test and quantitative PCR. Quantitative differences at RNA and protein levels were responsible for changes in viral production. To further characterize the mechanisms responsible for decreased rates of HIV replication in CD4-expressing cells we have treated the different cell lines, very early after HIV infection, with azidothymidine and soluble CD4, two antiviral agents that inhibit replication of HIV at different stages in the virus replicative cycle. Results from these experiments indicate that a cellular signal is mediated by the CD4 molecule, which negatively regulates the expression of viral DNA already present in such cells. This signal would be initiated following oligomerization of the CD4 molecule by the virus itself. Results from experiments with a CD4 construct containing mutations of the cysteine residues which are responsible for association of CD4 with p56lck demonstrate that p56lck is implicated in the transduction of the signal negatively regulating HIV replication.
Tremblay
M
M
Département de Microbiologie, Université Laval, Ste-Foy, Québec, Canada.
Meloche
S
S
Gratton
S
S
Wainberg
M A
MA
Sékaly
R P
RP
D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D008969:Molecular Sequence Data; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology / Q000382:microbiology; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000378:metabolism; D016662:Virus Integration; D014779:Virus Replication
D015704:CD4 Antigens; D012367:RNA, Viral; D014764:Viral Proteins; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)