Viral protease assay based on GAL4 inactivation is applicable to high- throughput screening in mammalian cells

Joseph F. Lawler, Solomon H. Snyder

Research output: Contribution to journalArticlepeer-review

Abstract

We present an assay for viral proteases that relies on the proteolytic cleavage of substrate leading to the dissociation of the yeast transcription factor GAL4. A consensus substrate for the cytomegalovirus protease is fused between the DNA binding and transactivating domains of GAL4. Proteolysis inactivates the transcription factor which drives a luciferase reporter system. The assay is performed in mammalian cells, has a robust signal-to- noise ratio, and assesses proteolysis in a physiologic context. A unique feature of the assay is its ability to detect inhibitors of viral replication that act on vital targets other than the protease.

Original languageEnglish (US)
Pages (from-to)133-138
Number of pages6
JournalAnalytical biochemistry
Volume269
Issue number1
DOIs
StatePublished - Apr 10 1999

Keywords

  • Cytomegalovirus
  • GAL4
  • Protease
  • Screen

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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