Figure 1

Interactions among the bacterial GroEL, aspartate aminotransferase (AST), and viral VP371 proteins. (A) Interaction between AST and GroEL. The cultures of GVE2-infected or non-infected thermophilic Geobacillus sp. E263 (wild-type, WT) were used for co-immunoprecipitation (Co-IP) with antibodies against GST, GST-MreB or GST-AST and used for GST pull down with GST, GST-MreB or GST-AST. The mutant of AST (∆ ast) was also included in the Co-IP assays. The antibodies used for IP were indicated at the top. The resulting Co-IP solutions were subsequently subjected to sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE; Coomassie staining) (up) and Western blot (down), respectively. The proteins used for GST pull down were presented at the top. For Western blot, the antibodies used were shown on the left. The arrow showed the protein identified using mass spectrometry. M, protein marker. (B) Interaction between VP371 and GroEL. The cultures of GVE2-infected or non-infected thermophilic Geobacillus sp. E263 were used for Co-IP with the VP371-specific, GST-MreB-specific or GST-specific antibodies and used for GST pull down. The resultant Co-IP solutions were subjected to SDS-PAGE (up) and Western blot (down), respectively. The mass spectrometric identification of protein was shown with an arrow. The proteins used for GST pull down were indicated at the top. M, protein marker. (C) Bacterial two-hybrid analysis of interactions among GroEL, aspartate aminotransferase and VP371 proteins. E. coli cells were co-transfected with recombinant plasmids as indicated at the top. The transformants were grown in agar plates containing the selective antibiotics TCK (tetracycline+chloramphenicol+ kanamycin) or CTCK (carbenicillin+tetracycline+ chloramphenicol+kanamycin). (D) Model of the linear interactions in the GroEL-aspartate aminotransferase-VP371 complex.