Classical swine fever virus NS3 is an IRES-binding protein and increases IRES-dependent translation

To get more evidences for understanding the role of NS3 in viral translation, we observed the promotive effect of CSFV NS3 on IRES-mediated translation by using dicistronic and monocistronic systems containing the precise segment comprising CSFV IRES. The results for affinity chromatography and UV-crosslinking assays indicated that NS3 bound CSFV IRES and that CSFV NS5A and NS5B could reduce the IRES-NS3 interaction. Further experiments showed that the NS5A also bound the IRES and that NS3 and NS5A bound the same binding sites of the IRES, suggesting that NS3 and NS5A competitively bind the same sites in IRES RNA sequence, thus hampering the interaction CSFV NS3 and IRES. But, CSFV NS5B was not found to interact with the IRES. The inhibitive effect of NS5B on binding of CSFV NS3 to IRES was supposed to result from the NS3–NS5B interaction which has been documented.