Genetic Selection of Poliovirus 2A(pro)-Binding Peptides

The yeast two-hybrid system has been used to identify mammalian clones that interact with poliovirus 2A proteinase (2A(pro)). Eight clones which encode previously unidentified human proteins were selected from a HeLa cell cDNA expression library. In addition, five clones encoding short peptides that interact with poliovirus 2A(pro) were also identified. The lengths of these peptides range from 6 to 30 amino acids, but all of them contain the Leu-X-Thr-Z motif (X represents any amino acid; Z represents a hydrophobic residue). This sequence is invariably located just at the carboxy terminus of each peptide. This approach raises the possibility of designing substrate analogue inhibitors of 2A(pro). Thus, two nonhydrolyzable peptides containing the Leu-X-Thr-Z motif prevented cleavage of eukaryotic initiation factor 4G by poliovirus 2A(pro) in vitro. A more general method for identifying peptides with antiproteinase activity is discussed.