title: T-cell receptor signaling enhances transcriptional elongation from latent HIV proviruses by activating P-TEFb through an ERK-dependent pathway.
abstract: Latent human immunodeficiency virus (HIV) proviruses are thought to be primarily reactivated in vivo through stimulation of the T-cell receptor (TCR). Activation of the TCR induces multiple signal transduction pathways, leading to the ordered nuclear migration of the HIV transcription initiation factors NF-κB (nuclear factor κB) and NFAT (nuclear factor of activated T-cells), as well as potential effects on HIV transcriptional elongation. We have monitored the kinetics of proviral reactivation using chromatin immunoprecipitation assays to measure changes in the distribution of RNA polymerase II in the HIV provirus. Surprisingly, in contrast to TNF-α (tumor necrosis factor α) activation, where early transcription elongation is highly restricted due to rate-limiting concentrations of Tat, efficient and sustained HIV elongation and positive transcription elongation factor b (P-TEFb) recruitment are detected immediately after the activation of latent proviruses through the TCR. Inhibition of NFAT activation by cyclosporine had no effect on either HIV transcription initiation or elongation. However, examination of P-TEFb complexes by gel-filtration chromatography showed that TCR signaling led to the rapid dissociation of the large inactive P-TEFb:7SK RNP (small nuclear RNA 7SK ribonucleoprotein) complex and the release of active low-molecular-weight P-TEFb complexes. Both P-TEFb recruitment to the HIV long terminal repeat and enhanced HIV processivity were blocked by the ERK (extracellular-signal-regulated kinase) inhibitor U0126, but not by AKT (serine/threonine protein kinase Akt) and PI3K (phosphatidylinositol 3-kinase) inhibitors. In contrast to treatment with HMBA (hexamethylene bisacetamide) and DRB (5,6-dichlorobenzimidazole 1-β-ribofuranoside), which disrupt the large 7SK RNP complex but do not stimulate early HIV elongation, TCR signaling provides the first example of a physiological pathway that can shift the balance between the inactive P-TEFb pool and the active P-TEFb pool and thereby stimulate proviral reactivation.
authors.affiliation: Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, 10900 Euclid Avenue, Room W200, Cleveland, OH 44106-4960, USA.
authors.forename: Young Kyeung
authors.initials: YK
authors.lastname: Kim
authors.forename: Uri
authors.initials: U
authors.lastname: Mbonye
authors.forename: Joseph
authors.initials: J
authors.lastname: Hokello
authors.forename: Jonathan
authors.initials: J
authors.lastname: Karn
mesh_terms: D002070:Butadienes / Q000494:pharmacology; D002843:Chromatin / Q000378:metabolism; D002867:Chromones / Q000494:pharmacology; D056906:Cyclin T / Q000378:metabolism; D042863:Cyclin-Dependent Kinase 9 / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006678:HIV / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D009025:Morpholines / Q000494:pharmacology; D050778:NFATC Transcription Factors / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D042925:Positive Transcriptional Elongation Factor B / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011533:Proviruses / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014775:Virus Activation / Q000187:drug effects; D017735:Virus Latency / Q000187:drug effects; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D002070:Butadienes; C503684:CCNT1 protein, human; D002843:Chromatin; D002867:Chromones; D056906:Cyclin T; D009025:Morpholines; D050778:NFATC Transcription Factors; D009570:Nitriles; D011948:Receptors, Antigen, T-Cell; D012261:Ribonucleoproteins; C113580:U 0126; D054322:tat Gene Products, Human Immunodeficiency Virus; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D042925:Positive Transcriptional Elongation Factor B; C467969:CDK9 protein, human; D042863:Cyclin-Dependent Kinase 9; D048049:Extracellular Signal-Regulated MAP Kinases; D013755:Tetradecanoylphorbol Acetate