1. Rpn11p mutation affects the recruitment of some other host factors into the tombusvirus replicase in yeast.
Rpn11p mutation affects the recruitment of some other host factors into the tombusvirus replicase in yeast. The Ded1p helicase was copurified with the p33 replication protein from wt or rpn11-14ts yeast. The FLAG-tagged p33 and FLAG-p92 were copurified from solubilized membranous fraction of yeast extracts using a FLAG affinity column (lanes 2 and 3 and lanes 5 and 6). The 6×His-tagged p33 and His6-p92 copurified using a FLAG-affinity column (lanes 1 and 4) were used as a negative control. (Top) Western blot analysis of 6×His-tagged Ded1p with anti-His antibody in the affinity-purified preparations. (Middle) Western blot analysis of the same samples as in the top portion but using anti-FLAG antibody. (Bottom) Western blot analysis of 6×His-tagged p33 and 6×His-Ded1 with anti-His antibody in the total protein extract from yeast expressing the shown proteins. Each experiment was repeated three times.
K. Reddisiva Prasanth et al. J. Virol. 2015;89: