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

Malgorzata Sudol, Jennifer L. Fritz, Melissa Tran, Gavin P. Robertson, Julie B. Ealy, Michael Katzman

Research output: Contribution to journalArticle

2 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)67-74
Number of pages8
JournalAntiviral Chemistry and Chemotherapy
Volume22
Issue number2
DOIs
StatePublished - Nov 1 2011

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Drug Discovery
  • Virology

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