Figure 7. hEndoV cleaved dsRNA as an I-RNase with TSN and ADAR2.

(a) ³²P-labelled 21-mer ssRNA containing inosine was incubated with hEndoV (lanes 2–5: 4 nM; lanes 1 and 6–9: 0 nM) and TSN (lanes 2–5 and 6–9: 0, 2, 10 and 20 nM). Cleavage sites were analysed by electrophoresis. A 21-mer ssRNA (right panel) was cleaved at the indicated positions (arrows, 12 mer). The graph showing yields of 12-mer products with increasing TSN concentration. (b) ³² P-labelled 21-mer ssRNA containing inosine was incubated with hEndoV (4 nM) and pdTp (lanes 2–8: 2.5, 0, 0.1, 0.5, 1, 2.5 and 5 mM). A 21-mer ssRNA (right panel) was cleaved at the indicated positions (arrows, 12-mer). The graph showing yields of 12-mer products with increasing pdTp concentration. (c) ³² P-labelled 49-mer dsRNA (lane 1) was incubated with hEndoV (lanes 2 and 4: 4 nM) and ADAR2 (lanes 3 and 4: 1 nM). Cleavage sites were analysed by electrophoresis. A 49-mer dsRNA (right panel) was cleaved at the indicated positions (arrows). (d) A model for the cleavage of dsRNA with ADARs, TSN and hEndoV.