Figure 4
Mapping the regions of VP1 and GM130 that are required for their interaction.
(a) The regions of VP1 interacting with GM130 were mapped by a Y2H assay. Truncation/deletion mutants of VP1 in pGBKT7 vector were used as the bait plasmids. These were tested for interaction with GM130 prey cloned into pGADT7 vector by α-Galactosidase assays performed on yeast colonies cotransformed with the prey and bait plasmids and selected on plates lacking adenine, histidine, leucine and tryptophan with X-α-Gal. “+” or gray sticks indicate VP1 prey constructs that interacted with GM130, while “–” or hollow bars indicate constructs that did not interact with GM130. (b) Molecular Modeling of CVB3 VP1 protein (GenBank accession number: M88483) on the basis of the published crystal structure of CVB3, as predicted by SWISS-Model server and Discovery Studio Visualizer software. The region of VP1 that binds to GM130 is colored in red. (c) The regions of GM130 interacting with VP1 were mapped by an Y2H assay. The prey proteins, the truncation/deletion mutants of GM130, were tested against the bait VP1. Shown are the results of Y2H α-Galactosidase assays performed on yeast colonies cotransformed with the prey and bait plasmids and selected on plates lacking adenine, histidine, leucine and tryptophan with X-α-Gal. “+” or gray sticks indicate GM130 prey constructs that interacted with VP1, while “–” or hollow bars indicate constructs that did not interact with VP1.
