Figure 5.

The requirement for adjacent single-stranded segments. The 5′-triphosphorylated substrates incubated with T170V and the positions of the resulting cleavages are shown schematically at the top of the panel. The first lacked the 3′ trailer, whereas the second had a complementary oligonucleotide overlapping the E3 site (see Supplementary Figure S4). As Figure 2, lanes 1–5 contain samples taken 0, 5, 15, 30 and 60 min after mixing substrate and enzyme, whereas lanes C₁ and C₂ correspond to substrate incubated without enzyme for 0 and 60 min. Lane M contains the products of cleaving full-length precursor with T170V. These are provided as size markers. Enzyme and initial substrate concentration and product staining as Figure 1.