Experimental Confirmation of the Role of the α1-α2 Hairpin PVY VPg in eIF4E Interaction.

In recent years, the eIF4E protein family has been actively studied as one of the major susceptibility factors for Solanaceae plants in relation to potyvirus infections. This makes eIF4E an attractive target for genome editing to generate resistant varieties. The viral protein VPg interacts with eIF4E family proteins. Measuring the in vitro affinity of VPg for different eIF4E isoforms has traditionally been difficult. The main reason for this is the high aggregation propensity of VPg. To overcome this, we used protein engineering to generate a chimeric construct based on a fluorescent protein. This approach greatly facilitated the measurement of affinity between two proteins using surface plasmon resonance (SPR). Our results confirm that the α1-α2 hairpin of PVY VPg is involved in complex formation with eIF4E. These findings provide new insights into the molecular mechanism of this protein-protein interaction.