FIG 5.
Reduced copurification of selected co-opted host proteins with the viral replicase from yeast coexpressing WW-domain protein. The viral replicase was purified through FLAG-tagged p33 from yeast extracts using a FLAG-affinity column. (A to G) For the top panel, Western blot analysis of copurified HA-tagged host protein expressed from the chromosomal location with anti-HA antibody was performed. For the middle panel, Western blot analysis of the purified Flag-with with anti-FLAG antibody was performed. For the bottom panel, Western blot analysis of total protein extract with anti-HA antibody to detect the total amount of the HA-tagged host protein expressed from the chromosomal location in each sample was performed. Lane 1, negative control based on yeast expressing His6-tagged p33 and the HA-tagged host protein; lane 2, yeast coexpressing Flag-p33 and the HA-tagged host protein; lane 3, yeast coexpressing Flag-p33 and the HA-tagged host protein in combination with WW-domain protein. All yeast strains actively replicate the TBSV repRNA (not shown). Each experiment was repeated two to four times.