Fig. 4
From: Soybean antiviral immunity conferred by dsRNase targets the viral replication complex
Rsv4 is a Mn2+-dependent dsRNase that associates with SMV replication machinery. a Activity of Rsv4. Purified Rsv4 protein or Rsv4 with a D99N substitution was incubated with 32P-labeled ssRNA (top), ssRNA hybridized with oligo DNA (middle), or dsRNA (bottom). Divalent cation was supplied as indicated. As controls, E. coli RNase H and S1 nuclease were used. RNA was detected by urea-PAGE followed by autoradiography. Positions of the substrate RNAs are indicated to the right by arrowheads. The bands marked by an asterisk migrated at the same speed as the ssRNA. b SMV P3-Myc is co-immunoprecipitated with Rsv4-FLAG. Proteins were detected by anti-FLAG and anti-Myc antibodies. c P3-Myc from SMV-L(Q1033K) is co-immunoprecipitated with Rsv4-FLAG. C(wt) indicates strain SMV-C. d Rsv4(D99N) associates with SMV RdRp in infected leaves. Membrane fractions of leaf homogenates (Membrane) and immunoprecipitates using anti-FLAG antibody after solubilization (FLAG-IP) were analyzed by 32P incorporation (RdRp activity) and western blotting (P3 and NIb). The source data of Fig. 4a-d are provided as a Source Data file
