Binding of USF1/2 and TFII-I (RBF-2) at conserved sites flanking the HIV-1 LTR enhancer is essential for reactivation from latency in T cells, with knockdown rendering provirus insensitive to cell signaling. We identified an interaction tripartite motif protein TRIM24, these factors were found be constitutively associated LTR. Similar effect depletion, loss TRIM24 impaired response deficiency did not affect recruitment RNA Pol II promoter, but inhibited transcriptional elongation, that was...

Expression of the HIV-1 genome by RNA Polymerase II is regulated at multiple steps, as are most cellular genes, including recruitment general transcription factors and control transcriptional elongation from core promoter. We recently discovered that tripartite motif protein TRIM24 recruited to Long Terminal Repeat (LTR) interaction with TFII-I causes stimulating association PTEF-b/ CDK9. Because required for stimulation LTR, we were surprised find IACS-9571, a specific inhibitor C-terminal...

Latent HIV-1 provirus represents the barrier toward a cure for infection and is dependent upon host RNA Polymerase (Pol) II machinery reemergence. Here, we find that inhibitors of Pol mediator kinases CDK8/19, Senexin A BRD6989, inhibit induction expression in response to latency-reversing agents T cell signaling agonists. These were found impair recruitment LTR. Furthermore, several latency reversal was impaired disruption CDK8 by shRNA or gene knockout. However, effects depletion did not...

ABSTRACT Current antiretroviral therapy for HIV-1 infection does not represent a cure as viral rebound inevitably occurs following discontinuation of treatment. The “block and lock” therapeutic strategy is intended to enforce proviral latency durably suppress viremic reemergence in the absence other intervention. transcription-associated cyclin-dependent protein kinases (tCDKs) are required expression from 5´ long-terminal repeat, but potential inhibiting these enforcing has been...

Abstract Expression of HIV-1 in response to T cell signaling requires TFII-I bound conserved sites flanking the LTR enhancer. Here we demonstrate that recruits tripartite motif protein TRIM24 by direct interaction. Constitutive interaction with was dependent upon TFII-I, and knockout impaired reactivation expression signaling. Loss did not affect recruitment RNA Pol II promoter, but inhibited transcriptional elongation, an effect associated decreased CTD S2 phosphorylation CDK9. Furthermore,...

Abstract Latent HIV-1 provirus represents a barrier towards cure for infection, but is dependent upon the host RNA Pol II machinery expression. We find that inhibitors of mediator kinases CDK8/19, Senexin A and BRD6989, inhibit induction expression in response to latency reversing agents T cell signaling agonists. These were found impair recruitment LTR. several reversal was impaired disruption CDK8 by shRNA or gene knockout. However, effects depletion did not entirely mimic CDK8/19 kinase...

HIV-1 provirus expression is controlled by signaling pathways that are responsive to T cell receptor engagement, including those involving Ras and downstream protein kinases. The induction of transcription from the LTR in response requires binding Ras-responsive element factor (RBF-2) conserved cis elements flanking enhancer region, designated RBE3 RBE1. RBF-2 composed minimally USF1, USF2, TFII-I factors. We recently determined regulates transcriptional elongation through recruitment...

Abstract HIV-1 latency is regulated by chromatin modifying enzymes, and histone deacetylase inhibitors (HDACi) were previously found to reactivate provirus expression. We report that of CBP/p300 acetyltransferases also cause reversal in T-cells. synergize with mechanistically diverse reversing agents reactivation. In contrast, inhibition impaired the HDACi SAHA, indicating contribute acetylation on LTR associated HDACi-mediated reversal. caused loss H3K27ac H3K4me3 from LTR, but did not...

Cdk8 of the RNA polymerase II mediator kinase complex regulates gene expression by phosphorylating sequence-specific transcription factors. This function is conserved amongst eukaryotes, but signals and mechanisms regulating activity phosphorylation its substrates are unknown. Full induction GAL genes in yeast requires transcriptional activator Gal4 Cdk8. We used a screen to identify regulators Cdk8-dependent on Gal4, from which we identified multiple mutants with defects TORC1 signaling....

Abstract Expression of the HIV-1 genome by RNA Polymerase II is regulated at multiple steps, as are most cellular genes, including recruitment general transcription factors and control transcriptional elongation from core promoter. We recently discovered that tripartite motif protein TRIM24 recruited to Long Terminal Repeat (LTR) interaction with TFII-I causes stimulating association PTEF-b/ CDK9. Because required for stimulation LTR, we were surprised find IACS-9571, a specific inhibitor...

Latent HIV-1 provirus represents a barrier towards cure for infection, but is dependent upon the host RNA Pol II machinery expression. We find that inhibitors of mediator kinases CDK8/19, Senexin A and BRD6989, inhibit induction expression in response to latency reversing agents T cell signaling agonists. These were found impair recruitment LTR. Knockdown CDK8 by shRNA, or gene knockout, also impaired HIV-1. Treatment CD4+ cells from infected people on ART with inhibited virus replication...

Abstract RBF-2, composed minimally of the factors USF1, USF2, and TFII-I, is cognate binding complex stringently conserved cis -elements on HIV-1 LTR, designated RBE3 RBE1. Mutations these elements prevent induction provirus in response to T cell signaling. However, function USF1 USF2 for this effect are relatively uncharacterized. Here, we find that deletion but not gene cells abrogates expression. Loss caused a reduction expression protein, an was associated with decreased mRNA abundance....

Abstract Antiretroviral therapy is not a cure for HIV-1 as viral rebound ensues immediately following discontinuation. The block and lock therapeutic strategy seeks to enforce proviral latency durably suppress viremic reemergence in the absence of antiretroviral therapy. Transcriptional Cyclin Dependent Kinase activity regulates LTR transcription, however, effect potential inhibiting these kinases enforcing remains unrecognized. Using newly developed small molecule inhibitors that are highly...

Abstract Cdk8 of the RNA Polymerase II mediator complex regulates genes by phosphorylating sequence specific transcription factors. Despite conserved importance for eukaryotic transcriptional regulation, signals regulating are unknown. Full induction yeast GAL requires phosphorylation Gal4 Cdk8, and we exploited this requirement growth gal3 on galactose to identify mutants affecting activity. Several from screen produced defects in TOR signaling. A mutant designated g al f our throttle ( gft...

Abstract Expression of the HIV-1 genome by RNA Polymerase II is regulated at multiple steps, as are most cellular genes, including recruitment general transcription factors and control transcriptional elongation from core promoter. We discovered that tripartite motif protein TRIM24 recruited to Long Terminal Repeat (LTR) interaction with TFII-I causes stimulating association PTEF-b/ CDK9. Because required for stimulation LTR, we were surprised find IACS-9571, a specific inhibitor C-terminal...

Abstract The conserved HIV-1 LTR cis elements RBE1/3 bind the factor RBF2, consisting of USF1/2 and TFII-I, are essential for reactivation by T cell signaling. We determined that TFII-I recruits tripartite motif protein TRIM24 to LTR, this interaction is required efficient expression in response signaling, similar effect depletion. Knockout did not affect recruitment RNA Pol II promoter, but inhibited transcriptional elongation, an was associated with decreased CTD S2 phosphorylation...