Modulation of host cell nuclear proteins that bind to HIV-1 trans-activation-responsive element RNA by phorbol ester.
The trans-activation-responsive (TAR) element located within the 5' untranslated region of HIV-1 mRNA is the cis-responsive element for Tat, the viral trans-activator protein. Several TAR RNA binding proteins (TRBPs) have been identified in the nuclear extract from HeLa cells as cellular factors required for a full Tat-mediated trans-activation. In this study, we have tried to identify TRBPs in human T cell line (MOLT-4) persistently infected with HIV-1. Nuclear extract from the infected MOLT-4 cells was analyzed by gel-retardation and uv cross-linking assays with radiolabeled TAR RNA probe. Two major complexes of TAR RNA with some cellular proteins were detected in the gel-retardation assay. As the components of these complexes, at least five TRBPs (p30, p37, p46, p50, and p56) showing specific binding to the TAR RNA were detected in the uv cross-linking assay. We also observed that the detectable levels of p37 and p50 in the infected MOLT-4 cells were greatly reduced after phorbol ester (TPA) treatment under the condition of which HIV-1 gene expression was increased by about fivefold. These results suggest that the modulation of TRBPs by some mitogenic stimuli such as TPA might have a role in the trans-activation of HIV-1 gene expression in vivo.
Masuda
T
T
Department of Biodefence and Medical Virology, Kumamoto University School of Medicine, Japan.
Harada
S
S
D001665:Binding Sites; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D015696:Gene Products, tat / Q000378:metabolism*; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015347:RNA Probes; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D009687:Nuclear Proteins; D015347:RNA Probes; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; D013755:Tetradecanoylphorbol Acetate