Development of a robust cell-based high-throughput screening assay to identify targets of HIV-1 viral protein R dimerization

Drug Des Devel Ther. 2013 May 24;7:403-12. doi: 10.2147/DDDT.S44139. Print 2013.


Targeting protein-protein interactions (PPI) is an emerging field in drug discovery. Dimerization and PPI are essential properties of human immunodeficiency virus (HIV)-1 proteins, their mediated functions, and virus biology. Additionally, dimerization is required for the functional interaction of HIV-1 proteins with many host cellular components. In this study, a bimolecular fluorescence complementation (BiFC)-based screening assay was developed that can quantify changes in dimerization, using HIV-1 viral protein R (Vpr) dimerization as a "proof of concept." Results demonstrated that Venus Vpr (generated by BiFC Vpr constructs) could be competed off in a dose-dependent manner using untagged, full-length Vpr as a competitor molecule. The change in signal intensity was measured quantitatively through flow cytometry and fluorescence microscopy in a high content screening assay. High content imaging was used to screen a library of small molecules for an effect on Vpr dimerization. Among the tested molecules, a few of the small molecules demonstrate an effect on Vpr dimerization in a dose-dependent manner.

Keywords: BiFC; HIV-1 Vpr; dimerization; drug targets; protein-protein interaction.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Gene Products, vpr / chemistry*
  • HeLa Cells
  • High-Throughput Screening Assays*
  • Humans
  • Kinetics
  • Peptide Library
  • Protein Multimerization*


  • Gene Products, vpr
  • Peptide Library