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 language | English (US) |
---|---|
Pages (from-to) | 133-138 |
Number of pages | 6 |
Journal | Analytical biochemistry |
Volume | 269 |
Issue number | 1 |
DOIs | |
State | Published - Apr 10 1999 |
Keywords
- Cytomegalovirus
- GAL4
- Protease
- Screen
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology