FIG. 3.
(A) Schematic representation of MBP fusion proteins containing wild-type UL34 and mutant UL34 with alanine substitution(s) at Thr-195 and/or Ser-198. The amino acid sequence of UL34 residues 186 to 208 is shown. Sites with the consensus sequence for phosphorylation by Us3 are underlined. (B) CBB-stained and autoradiographic images of phosphorylated UL34. Purified MBP-UL34 (lanes 1 and 2), MBP-UL34T195A (lanes 3 and 4), MBP-UL34S198A (lanes 5 and 6), and MBP-UL34T195A-S198A (lanes 7 and 8) were incubated in kinase buffer containing [γ-32P]ATP and purified GST-Us3 (lanes 1, 3, 5, and 7) or GST-Us3K220M (lanes 2, 4, 6, and 8), separated on a denaturing gel, and stained with CBB (upper panel). The lower panel shows an autoradiograph of the gel in the upper panel.