An RIG-I-Like RNA Helicase Mediates Antiviral RNAi Downstream of Viral siRNA Biogenesis in Caenorhabditis elegans (original) (raw)
Figure 2
Molecular and functional characterization of the drh-1 and drh-2 genes.
(A) Accumulation of the FR1gfp replicon RNAs in genetic mutant worm strains carrying the same FR1gfp transgene array. Total RNA was also analyzed from wildtype N2 worms with (N2) and without the FR1gfp transgene (N2*) 48 hours after induction of the replicon replication (h.a.i.). (B) Induction of the RNAi immunity by the replicon in a worm integrant different from that analyzed in (A). (C) Molecular structures and genetic lesions of the drh-1 and drh-2 genes. (D) Accumulation of (-) viral siRNAs in single knockout worm mutants 48 hours after induction of the replicon replication. 45 µg of total small RNAs was loaded in each lane. A combination of 18 32P end-labeled DNA oligos corresponding to eGFP coding sequence in tandem was used as the probe for viral siRNA detection. The same filters were probed for miR-58 after stripping as the loading control. (E) Detection of drh-1 and drh-2 transcripts before and after induction of the replicon replication. Two independent tests were analyzed for each strain. (F) Time course analysis of the accumulation of the replicon RNAs in wildtype and mutant worms 2, 6, and 16 h.a.i. Total RNA extracted from FR1gfp rde-1 worms 48 h.a.i. was loaded as a control (lane rde-1*). Methylene blue staining of total RNA was provided to show equal loading.