Inactivation of the Host Lipin Gene Accelerates RNA Virus Replication through Viral Exploitation of the Expanded Endoplasmic Reticulum Membrane
Figure 7
Enhanced activity of the affinity-purified tombusvirus replicase assembled in CFE from pah1Δ nem1Δ yeast.
(A) Scheme of the tombusvirus replicase assay. CFEs were prepared from wt and pah1Δ nem1Δ yeasts, followed by the addition of purified recombinant tombusvirus p33 and p92pol replication proteins and DI-72 (+)repRNA in the presence of rATP and rGTP. The in vitro assays were programmed with RI/III (−)RNA in the presence of rATP/rCTP/rGTP and 32P-rUTP. (B) Top panel: Representative denaturing gel of 32P-labeled RNA products synthesized by the purified tombusvirus replicase in vitro. Each experiment was repeated. Bottom panel: Western blot analysis of p33 in the shown replicase samples.