Table 2.
Summarized characterization of the kinase-specific parameters of mutant versions of pUL97
++, positive reaction, very strong signal; +, positive reaction, clearly detectable signal; ±, weak signal; −, negative.
| Ectopic expression of fragments or replacement mutants | Kinase-specific parameters |
Phenotype of pUL97 | ||
|---|---|---|---|---|
| Self-interactiona | Kinase activityb | Cyclin B1 bindingc | ||
| 1–707 | ++ | ++ | + | pUL97 full-length, wildtype activity |
| 1–706 | ++ | + | + | C-terminal truncation, catalytically active |
| 1–702 | + | − | ± | C-terminal truncation, catalytically inactive |
| R702A/L704A | ++ | ++ | + | Mutation of putative cyclin-docking motif |
| K355M | ++ | − | − | Catalytically inactive mutant (ATP-binding site) |
| 1–707 + MBV | ++ | − | ± | pUL97 full-length, activity inhibited |
| S483A/S485A | ++ | − | − | Mutation of serine residues in putative T-loop |
| S483A | ++ | − | + | Mutation of serine residues in putative T-loop |
| S485A | ++ | + | + | Mutation of serine residues in putative T-loop |
| S483D/S485D | ++ | − | − | Mutation of serine residues in putative T-loop |
| S483E/S485E | + | − | − | Mutation of serine residues in putative T-loop |
| Q382A/H406A/R451A | ± | − | − | Mutation of putative cyclin-binding interface |
| Q382A/H406A/R645A | ± | − | − | Mutation of putative cyclin-binding interface |
| Q382A/H451A/R645A | ± | + | + | Mutation of putative cyclin-binding interface |
| Gln-382/His-406/His-448/Arg-451/Asp-490/Ser-643/Arg-645 | + | − | − | Mutation of putative cyclin-binding interface |
a Determined by CoIP analysis using two different tagged versions of pUL97.
b Determined by in vitro kinase assay measuring autophosphorylation and histone phosphorylation.
c Determined by CoIP analysis (interaction profiles of N-terminal truncations published in Steingruber et al. (16).