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. 2018 May;24(5):721–738. doi: 10.1261/rna.065037.117

FIGURE 2.

FIGURE 2.

Recombinant Grc3/Las1 requires a γ-phosphate for RNA phosphorylation. (A) RNA phosphorylation activity of Grc3/Las1 (4 µM) with ssRNA (5 µM) in the absence (x) and presence of ATP, ADP, and ADPnP at 1 mM. (B) ATP/ADP competition assay. Grc3/Las1 (4 µM) was incubated with 10 µM fluorescently labeled ssRNA with constant ATP (30 µM) and a concentration series of ADP (0–5 mM). (C) ATP/ADPnP competition assay. Grc3/Las1 (4 µM) was incubated with 10 µM fluorescently labeled ssRNA with constant ATP (30 µM) and a concentration series of ADPnP (0–5 mM). The average and standard deviation were measured from three independent replicates. (*) P < 0.02; (**) P < 0.005; (***) P < 2 × 10−4; (****) P < 2 × 10−5 were calculated from two-tailed Student's t-tests.