1. Effect of phospholipids on the in vitro RdRp activity of the p92-Δ167N RdRp protein.
Effect of phospholipids on the in vitro RdRp activity of the p92-Δ167N RdRp protein. (A) PAGE analysis of the 32P-labeled RNA products obtained in an in vitro assay with purified p92-Δ167N RdRp protein in the presence of purified Ssa1 Hsp70 (13 pmol) and DI-mini (+)RNA template. In addition, samples also contained various phospholipids as shown (70 μM). See further details in Fig. 1. Each experiment was repeated two times. (B) Denaturing PAGE analysis of the 32P-labeled RNA products obtained in an in vitro assay with FLAG-affinity-purified CNV replicase in the presence of the buffer, DMSO, or the shown phospholipids (70 μM). The template used was DI-72 (−)RNA, which leads to the production of de novo-initiated full-length (top band, marked with an arrowhead and quantified) and internally initiated shorter products. Note that samples contained 0.1% detergent required for the purification of the replicase preparation.
Judit Pogany, and Peter D. Nagy J. Virol. 2015;89: