The infection of cattle with Foot and Mouth Disease Virus (FMDV) causes not only great economic damage but great popular panic, e.g., the 2001 outbreak in Great Britain which is estimated to have cost the UK $16 billion. Future outbreaks arising from either natural causes or as a result of bioterrorism need to be dealt with quickly, effectively and cheaply to prevent rapid spread. There are currently no approved anti- FMDV drugs for the treatment or prevention of FMD. FMDV utilizes a highly unusual form of protein translation termed StopGo to separate its coat proteins from those proteins required for replication. StopGo is dependent on a highly conserved signature sequence, DxExNPG-P, where DxExNPG are the last 7 amino acids of a protein termed 2A and the P is the first amino acid of a protein termed 2B. Ribosomes translate 2A conventionally but then stop, releasing the 2A product, before immediately restarting to make 2B. The viral RNA that encodes the StopGo signature has none of the conventional signals for ribosome starting and stopping at the 2A-2B junction. Instead, the nascent 2A protein sequence growing within the ribosome must interact within the ribosome to prompt the Stop and the Go. The goal of this proposal is to identify inhibitors of FMDV StopGo that specifically target the resumption of translation on P (Go) rather than the termination at G (Stop). We plan to: (1) Identify novel small molecule inhibitors of FMDV StopGo by high throughput screening (HTS) using a dual luciferase enzyme assay against the MLSCN (Molecular Libraries Screening Center Network) compound collection. (2) Characterize and optimize the FMDV StopGo inhibitors identified from HTS. (3) Validate the antiviral activity of these compounds using cells lines infected with FMDV.