A small molecule screen in yeast identifies inhibitors targeting protein-protein interactions within the vaccinia virus replication complex

► We report the use of a forward duplex yeast two-hybrid assay to screen molecules. ► Two molecules interfere with protein interactions within the VACV replication complex. ► Molecules inhibit VACV in vitro and display a similar antiviral activity than CDV. ► Molecules inhibit CPXV but not HSV-1, thus demonstrating a good specificity. ► We showed that, as expected, the two molecules interfere with VACV DNA synthesis.