Extended Data Figure 5: Using virus engineering to discern Drosha’s antiviral mechanism.
From: RNase III nucleases from diverse kingdoms serve as antiviral effectors
a, Schematic of the SINV replicon encoding Gaussia luciferase in place of the structural polyprotein used in Fig. 3e–g. b, Schematic of the SINV temperature-sensitive mutant (SIN-RdRpts). Star denotes ts point mutant. c, NoDice and RNaseIII−/− cells were infected with virus depicted in b, at an MOI of 10 and incubated at 40 °C, a temperature at which the mutant viral RdRp is completely inactive. Levels of genomic (g) SINV RNA were determined by qPCR at the indicated times after infection. Data are representative of two independent experiments where each condition was done in triplicate; error bars, s.d. d, Schematic of the SINV encoding firefly luciferase in the nsP3 region and an inactive RdRp (SIN-nsP3Luc) e, Graph depicts levels of in vitro translation of firefly luciferase produced from virus in d, in the presence of membrane fractions from control or Drosha-2A. The data shown are the average of three independent experiments; error bars, s.d.
