TABLE 5.
Functional characterization of DNMT3A dimer interface mutants with DNMT3L, showing DNMT3L restores processivity in the R882H mutant
Values were determined as stated in Table 1 but with DNMT3L preincubated with enzyme for 1 h at 37 °C in reaction buffer with AdoMet at a 1:1 ratio of DNMT3A to DNMT3L. Fold stimulation were determined by the fold change in methyl group transferred in 1 h comparing with and without DNMT3L.
| Activation | Fold ↑ | koff | Fold ↑ | Processivity | Fold ↓ | Processive | |
|---|---|---|---|---|---|---|---|
| min−1 | n½ | ||||||
| WT + 3L | 5.4 ± 0.2 | 1 | 0.17 ± 0.01 | 1.0 | 154 ± 45.2 | 1.0 | Yes |
| H873A + 3L | 3.7 ± 0.5 | 0.68 | 2.25 ± 0.25 | 13.4 | 6.4 ± 2.19 | 24.0 | Some |
| H873R + 3L | 3.1 ± 0.6 | 0.58 | |||||
| W860A + 3L | 2.9 ± 0.5 | 0.54 | 2.16 ± 0.34 | 12.9 | |||
| N879A + 3L | 2.2 ± 0.2 | 0.41 | |||||
| R882H + 3L | 8.9 ± 0.8 | 1.65 | 0.43 ± 0.07 | 2.6 | 54 ± 8.6 | 2.9 | Yes |
| D876G + 3L | 3.7 ± 0.2 | 0.68 | |||||
| R885A + 3L | No activity |