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. 2014 Apr 17;10(4):e1004051. doi: 10.1371/journal.ppat.1004051

Figure 4. AtRH2 and AtRH5 can unwind short partial RNA/RNA duplex within the RIII(−) REN in vitro.

Figure 4

(A) Schematic representation of the partial RNA/RNA duplex used in the strand separation assay. The unlabeled template consists of DI-72 (−)repRNA and a complementary RIII(+) RNA, which anneals to the DI-72 (−)repRNA and forms a 82 nt duplex as shown. (B) Representative native gel of 32P-labeled RNA products after the in vitro strand separation assay. Strand separation assay with a partial RNA/RNA duplex shows the unwinding activity of AtRH2 and AtRH5, Fal1p and Dbp3p (1.0 µg amount) in the presence of ATP. The DDX3-like Ded1p and AtRH20 helicases show only partial unwinding activities under these conditions. Quantification of the RNA/RNA duplex is done with a Phosphorimager.