title: Evaluation of a system to screen for stimulators of non-specific DNA nicking by HIV-1 integrase: application to a library of 50,000 compounds.
abstract: BACKGROUND
  In addition to activities needed to catalyse integration, retroviral integrases exhibit non-specific endonuclease activity that is enhanced by certain small compounds, suggesting that integrase could be stimulated to damage viral DNA before integration occurs.
  
  
  METHODS
  A non-radioactive, plate-based, solution phase, fluorescence assay was used to screen a library of 50,080 drug-like chemicals for stimulation of non-specific DNA nicking by HIV-1 integrase.
  
  
  RESULTS
  A semi-automated workflow was established and primary hits were readily identified from a graphic output. Overall, 0.6% of the chemicals caused a large increase in fluorescence (the primary hit rate) without also having visible colour that could have artifactually caused this result. None of the potential stimulators from this moderate-size library, however, passed a secondary test that included an inactive integrase mutant that assessed whether the increased fluorescence depended on the endonuclease activity of integrase.
  
  
  CONCLUSIONS
  This first attempt at identifying integrase stimulator compounds establishes the necessary logistics and workflow required. The results from this study should encourage larger scale high-throughput screening to advance the novel antiviral strategy of stimulating integrase to damage retroviral DNA.
authors: Department of Medicine, Penn State College of Medicine, Milton S Hershey Medical Center, Hershey, PA, USA.
authors: Malgorzata
authors: M
authors: Sudol
authors: Jennifer L
authors: JL
authors: Fritz
authors: Melissa
authors: M
authors: Tran
authors: Gavin P
authors: GP
authors: Robertson
authors: Julie B
authors: JB
authors: Ealy
authors: Michael
authors: M
authors: Katzman
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology*; D053904:DNA Breaks, Single-Stranded; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005453:Fluorescence; D019427:HIV Integrase / Q000235:genetics*; D054852:Small Molecule Libraries; D016662:Virus Integration / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents; D054852:Small Molecule Libraries; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1