qHTS Assay for Inhibitors of the HIV-1 protein Vpr
Human immunodeficiency virus type 1 (HIV-1), the causative agent of AIDS, contains a protein called viral protein R (Vpr) that plays an important role in the viral life cycle and the pathogenesis of HIV-1. ..more
BioActive Compounds: 750
Human immunodeficiency virus type 1 (HIV-1), the causative agent of AIDS, contains a protein called viral protein R (Vpr) that plays an important role in the viral life cycle and the pathogenesis of HIV-1.
Vpr plays multiple roles during the viral life cycle and displays several distinct activities in host cells. These Vpr specific activities include cytoplasmic-nuclear shuttling, induction of cell cycle G2 arrest and cell killing. The cytoplasmic-nuclear shuttling is believed to participate in nuclear transport of the proviral integration complex (PIC). The cell cycle G2 arrest induced by Vpr is thought to suppress human immune functions by preventing T cell clonal expansion and to provide an optimized cellular environment for maximal levels of viral replication. In addition, Vpr induces apoptosis, which may contribute to the depletion of CD4 T cells in HIV-infected patients. Since the Vpr-specific activities have been linked to such clinical manifestation of AIDS as activation of viral replication, suppression of host immune responses and depletion of CD4+ T-lymphocytes, identification of new molecular probes that can inhibit the Vpr activities could potentially provide a new approach to reduce Vpr-mediated detrimental effects in HIV-infected patients, and thus prolong patient's lives. Currently, there are no anti-Vpr drugs.
A fission yeast (Schizosaccharomyces pombe) model system for the Vpr studies has been developed which measures HIV-1 Vpr-induced cell death. This assay will be used for to screen the NIH Molecular Libraries Small Molecule Repository (MLSMR) identify small molecule HIV-1 Vpr inhibitors.
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]
MLPCN Grant: NS063880
Assay Submitter (PI): Richard Zhao, University of Maryland
The fission yeast RE007 strain was used, which carries a single integrated copy of HIV-1 vpr gene in the yeast chromosome. The starting culture was frozen stock 100ul of yeast RE007 in 150ml of *PMG+AL medium, Incubation in 30 degree plus 99% humidity for 18 hours. Next day dispensed 2.5ul of PMG+AL medium in each well to 1536 well white solid plate, then 23nL compound were delivered to each well using a pin tool. Check OD for overnight culture and add cell density at 0.01 for each well, incubation in 30 degree plus 99% humidity for 24 hours. After add 5 ul of *BacTiter/Glo detect reagent to each well at room temperature for 5 minutes then to red out results with Viewlux CCD camera.
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
* Activity Concentration. ** Test Concentration.
Data Table (Concise)