TABLE 2.
RNA binding assay of catalytic mutantsa
| Strain or catalytic mutant | Fluorescence polarization GpppG-SUDV12 (Kd) (μM) |
|---|---|
| WT | 0.740 (±0.108) |
| K1813A | 4.6 (±0.943) |
| D1924A | 0.581 (±0.05) |
| K1959A | ND |
| E1994A | 1.3 (±0.206) |
a
RNA-binding assays were performed with fluorescence polarization using a 13-mer SUDV-specific capped RNA (GpppG-SUDV12) labeled at the 3′ end with pCp-Cy5 and wild-type (WT) SUDV MTase+CTD and the indicated catalytic mutant MTase+CTD proteins. The dissociation coefficient (Kd) was calculated by site-specific binding regression with Hill slope. Data are the mean (± standard deviation) (n = 3). ND, not determined.