Immunodeficiency virus rev trans-activator modulates the expression of the viral regulatory genes

Malim, Michael H.; Hauber, Joachim; Fenrick, Randy; Cullen, Bryan R.

doi:10.1038/335181a0

Cited by 371 publications

(345 citation statements)

References 25 publications

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“…3A). It is reported that Rev, a viral protein, is associated with positive modulation of virion production through up-regulation of mRNA transport into the cytoplasm by interaction with its RNA target, the rev-responsive element (50,51). Thus, this dichotomous effect of HU on HIV activation suggests that HU/IL-6 may enhance the Rev/rev-responsive element (RRE) posttranscriptional pathway, whereas HU/TNF-␣ may interfere with the Rev/RRE elements.…”

Section: Discussionmentioning

confidence: 99%

Hydroxyurea and Interleukin-6 Synergistically Reactivate HIV-1 Replication in a Latently Infected Promonocytic Cell Line via SP1/SP3 Transcription Factors

Oguariri

Brann

Imamichi

2007

Journal of Biological Chemistry

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The existence of viral latency limits the success of highly active antiretroviral therapy. With the therapeutic intention of reactivating latent virus to induce a cure, in this study we assessed the impact of cell synchronizers on HIV gene activation in latently infected U1 cells and investigated the molecular mechanisms responsible for such effect. Latently infected U1 cells were treated with 10 drugs including hydroxyurea (HU) and HIV-1 replication monitored using a p24 antigen capture assay. We found that HU was able to induce HIV-1 replication by 5-fold. HU has been used in the clinical treatment of HIV-1-infected patients in combination with didanosine; therefore, we investigated the impact of HU on HIV-1 activation in the presence of the proinflammatory cytokines, interleukin 6 (IL-6) and tumor necrosis factor-␣ (TNF-␣). IL-6 or TNF-␣ alone induced HIV replication by 18-and ϳ500-fold, respectively. Of interest, in the presence of HU, IL-6-mediated HIV-1 activation was enhanced by >90-fold, whereas TNF-␣-mediated activation was inhibited by >30%. A reporter gene assay showed that HU and IL-6 synergized to activate HIV promoter activity via the Sp1 binding site. Electrophoretic mobility shift and supershift assays revealed increased binding of the Sp1 and Sp3 transcription factors to this region. Western blot analysis showed that HU and IL-6 co-stimulation resulted in increased levels of Sp1 and Sp3 proteins. In contrast, treatment with HU plus TNF-␣ down-regulated the expression of NF-B. These findings suggest that Sp1/Sp3 is involved in controlling the HU/IL-6-induced reactivation of HIV-1 in latently infected cells.

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Section: Discussionmentioning

confidence: 99%

Hydroxyurea and Interleukin-6 Synergistically Reactivate HIV-1 Replication in a Latently Infected Promonocytic Cell Line via SP1/SP3 Transcription Factors

Oguariri

Brann

Imamichi

2007

Journal of Biological Chemistry

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“…The CXCR4-tropic HIV-1 molecular clone pR8 lacking env (pR8⌬Env) was obtained from Dr. D. Trono (University of Geneva, Switzerland). The pcRev plasmid (34) and the pMM310 plasmid expressing BlaM-Vpr (35) were obtained from the AIDS Research and Reference Reagent Program, National Institutes of Health.…”

Section: Methodsmentioning

confidence: 99%

Distinct Requirements for HIV-Cell Fusion and HIV-mediated Cell-Cell Fusion

Kondo

Marin

Kim

et al. 2015

Journal of Biological Chemistry

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“…HIV-1 pseudoviruses bearing HXB2, Bal26, and R3A envelope glycoproteins and the b-lactamase-Vpr chimera (BlaM-Vpr) were produced by cotransfecting HEK293T/17 cells with the HIV-based pR8DEnv packaging vector, pMM310 plasmid expressing BlaM-Vpr, 47 pcRev, 48 and the vector encoding HXB2 Env 49 or Bal26 (a gift from P. Clapham, University of Massachusetts, Worchester, MA) or R3A (a gift from Dr. J. Hoxie, University of Pennsylvania, Philadelphia, PA), using the jetPRIME Ò transfection reagent (VWR, Radnor, PA). The transfection medium was replaced with fresh DMEM/ 10% FBS after 12 h and cells were cultured for 36 h, after which time the virus-containing culture medium was collected, passed through a 0.45-mm filter, aliquoted, and stored at -80°C.…”

Section: Virus Productionmentioning

confidence: 99%

High-Throughput HIV–Cell Fusion Assay for Discovery of Virus Entry Inhibitors

Marin

Giroud

et al. 2015

ASSAY and Drug Development Technologies

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HIV-1 initiates infection by merging its envelope membrane with the target cell membrane, a process that is mediated by the viral Env glycoprotein following its sequential binding to CD4 and coreceptors, CXCR4 or CCR5. Although HIV-1 fusion has been a target for antiviral therapy, the virus has developed resistance to drugs blocking the CCR5 binding or Env refolding steps of this process. This highlights the need for novel inhibitors. Here, we adapted and optimized an enzymatic HIV-cell fusion assay, which reports the transfer of virus-encapsulated b-lactamase into the cytoplasm, to high-throughput screening (HTS) with a 384-well format. The assay was robustly performed in HTS format and was validated by the pilot screen of a small library of pharmacologically active compounds. Several hits identified by screening included a prominent cluster of purinergic receptor antagonists. Functional studies demonstrated that P2X1 receptor antagonists selectively inhibited HIV-1 fusion without affecting the fusion activity of an unrelated virus that enters cells through an endocytic route. The inhibition of HIV-cell fusion by P2X1 antagonists was not through downmodulation of the cell surface expression of CD4 or coreceptors, thus implicating P2X1 receptor in the HIV-1 fusion step. The ability of these antagonists to inhibit viruses regardless of their coreceptor (CXCR4 or CCR5) preference indicates that fusion is blocked at a late step downstream of coreceptor binding. A future large-scale screening campaign for HIV-1 fusion inhibitors, using the above functional readout, will likely reveal novel classes of inhibitors and suggest potential targets for antiviral therapy.

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Immunodeficiency virus rev trans-activator modulates the expression of the viral regulatory genes

Cited by 371 publications

References 25 publications

Hydroxyurea and Interleukin-6 Synergistically Reactivate HIV-1 Replication in a Latently Infected Promonocytic Cell Line via SP1/SP3 Transcription Factors

Hydroxyurea and Interleukin-6 Synergistically Reactivate HIV-1 Replication in a Latently Infected Promonocytic Cell Line via SP1/SP3 Transcription Factors

Distinct Requirements for HIV-Cell Fusion and HIV-mediated Cell-Cell Fusion

High-Throughput HIV–Cell Fusion Assay for Discovery of Virus Entry Inhibitors

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