TABLE 2.
IntDOT mutants isolated in this study
| Domain | IntDOT mutanta | Base changeb | Mutant activityc |
|||
|---|---|---|---|---|---|---|
| E. coli in vivo integration | Binding | Cleavage | Ligation | |||
| None | WT | ++ | ++ | ++ | ++ | |
| pBAD18 | − | − | − | − | ||
| N | R13C | CGT→TGT | − | + | ++ | ++ |
| S38N | AGC→AAC | + | + | ++ | ++ | |
| V95 M | GTG→ATG | − | + | ++ | ++ | |
| G101R | GGA→AGA | + | + | ++ | ++ | |
| CB | T184I (2 isolates) | ACC→ATC | − | + | + | + |
| P209L | CCG→CTG | + | + | + | + | |
| CAT | T256I | ACC→ATC | − | − | − | − |
| W280 | TGG→TAG | − | − | − | − | |
| R285H | CGC→CAC | − | +/++ | −/+ | − | |
| R285A (site directed) | CGC→GCC | − | ++ | − | −/+ | |
| R285D (site directed) | CGC→GAC | − | ++ | − | − | |
| R285K (site directed) | CGC→AAA | − | ++ | −/+ | − | |
| R295H | CGC→CAC | − | ++ | ++ | ++ | |
| V319I (2 isolates) | GTC→ATC | + | + | + | + | |
| C325Y (2 isolates) | TGC→TAC | − | − | − | − | |
| L326F | CTT→TTT | − | − | − | − | |
| R330H | CGT→CAT | − | − | − | − | |
| S347N | AGC→AAC | − | + | − | − | |
| R348H | CGC→CAC | − | + | − | − | |
| A352T | GCC→ACC | − | ++ | −/+ | ++ | |
| T354I | ACC→ATC | + | ++ | ++ | ++ | |
| T365 M (2 isolates) | ACG→ATG | − | ++ | ++ | ++ | |
| S367N | AGC→AAC | + | ++ | −/+ | + | |
| S368F | TCC→TTT | + | ++ | ++ | ++ | |
| G371E | GGA→GAA | − | −/+ | − | − | |
| H372Y | CAC→TAC | − | ++ | − | − | |
| A382V | GCG→GTG | − | ++ | ++ | ++ | |
| L389F (2 isolates) | CTC→TTC | + | ++ | ++ | ++ | |
| Y381F (site directed) | TAC→TTC | − | ++ | − | ++ | |
a
The first letter is the amino acid in wild-type IntDOT, followed by the position and the amino acid substituted for the wild-type amino acid. If only the first letter is given, the mutation creates a nonsense codon.
b
Base pair changes are shown in boldface.
c
++, +, and − are explained in Materials and Methods.