Figure 5.

N-terminus of eIF4G (eIF4G-N) interacts with RBSDV P8 in yeast and in planta. (A) Yeast two-hybrid assays show that eIF4G-N interacts with the RBSDV P8 protein. The plasmid combinations indicated on the right side of the image panel were cotransformed into yeast strain Y2H Gold, and the yeast cells were spotted on SD/-Trp/-Leu and SD/-Trp/-Leu/-His/-Ade selective medium with X-a-gal and grown at 30 °C for 3–4 d. Positive control, full-length pGADT7–53 and pGBKT7-T7. (B) The luciferase complementation imaging assay indicates that eIF4G-N interacts with the RBSDV P8 protein in planta. The plasmid combinations indicated on the left side of the image panel were coexpression Nicotiana benthamiana leaves by Agrobacterium-mediated infiltration. After 48 h, N. benthamiana leaves were sprayed with Lucifer, and a fluorescence signal appeared where eIF4G-N-nLuc or eIF4G^mu-N-nLuc was coexpressed with cLuc-P8. eIF4G^mu, eIF4G mutant protein generated by CRISPR/Cas9.