Fig 1.

Binding of UL84 and UL84 mutants to UL44 in vitro. (A) Schematic of UL84 and UL84 mutants used in GST pulldown assays. The residues of UL84 at which truncations have been made are noted above the figure. The name of each mutant is noted to the right of the figure. (B and C) GST pulldown assays were performed where GST or GST-UL44ΔC290 fusion proteins were incubated with radiolabeled UL84 or UL84 mutants shown in panel A produced by in vitro transcription/translation and passed over a glutathione column. The radiolabeled and GST proteins used in each reaction are noted above and below the figure, respectively. The input (B) and protein eluted by glutathione (C) from each reaction are shown. (D) Protein products of in vitro transcription/translation in reaction mixtures containing no protein expression vector (lane 1) or an expression vector encoding UL84(1–68) (lane 2) (from panel A). (E) GST pulldown assays were performed where GST, GST-UL44ΔC290, or GST-US11 fusion protein was incubated with radiolabeled UL84(1–68) and passed over a glutathione column. The GST proteins used in each reaction mixture are noted above the figure. Lane 1, input protein; lanes 2 to 4, protein eluted by glutathione from each reaction. The position of the approximately 10-kDa form of UL84(1–68) is indicated with an arrow in panels D and E. The position of molecular mass markers in panels B to E are indicated to the left of the figure.