Components of the Vps34p PI3K complex are essential host factors for tombusvirus replication in yeast.

(A) Deletion of selected yeast genes, which are parts of Vps34 complexes, inhibits TBSV replication in yeast. Top panel: Northern blot analysis of TBSV repRNA using a 3’ end specific probe shows the reduced accumulation of repRNA in the shown yeast strains in comparison with the wt yeast strain. Viral proteins His6-p33 and His6-p92pol were expressed from plasmids from the GAL1 promoter, while DI-72(+) repRNA was expressed from the GAL10 promoter. Middle panel: Northern blot with 18S ribosomal RNA specific probe was used as a loading control. Bottom images: Western blot analysis of the level of His6-p33 and His6-p92pol with anti-His antibody. (B) Deletion of selected yeast genes, which are parts of Vps34 complexes, inhibits CIRV replication in yeast. See further details in panel A. Each experiment was repeated three times.