Application of a cell-based protease assay for testing inhibitors of picornavirus 3C proteases.

ABSTRACT: Proteolytical cleavage of the picornaviral polyprotein is essential for viral replication. Therefore, viral proteases are attractive targets for anti-viral therapy. Most assays available for testing proteolytical activity of proteases are performed in vitro, using heterologously expressed proteases and peptide substrates. To deal with the disadvantages associated with in vitro assays, we modified a cell-based protease assay for picornavirus proteases. The assay is based on the induction of expression of a firefly luciferase reporter by a chimeric transcription factor in which the viral protease and cleavage sites are inserted between the GAL4 binding domain and the VP16 activation domain. Firefly luciferase expression is dependent on cleavage of the transcription factor by the viral protea

SUBMITTER: van der Linden L 

PROVIDER: S-EPMC7113757 | biostudies-literature | 2014 Mar

REPOSITORIES: biostudies-literature