FIG. 6.

ICP27 interacts with TAP/NXF1 in vitro, and Aly/REF is not required to bridge the interaction. (A) A schematic diagram of ICP27 shows the leucine-rich region (LRR), the NLS, the RGG box RNA binding domain, three predicted KH domains, and a zinc finger-like motif (CCHC). The positions of the deletion mutations that were used in the in vitro binding assays are shown. (B) GST binding assays were performed with GST-TAP and in vitro-translated wild-type (WT) ICP27 and ICP27 mutants ΔLRR, D2ΔS5, S5, R1, ΔNLS, H17, and ΔC. Aly/Ref was included as a positive control, and luciferase (Luc) was included as a negative control. The in vitro binding assays were performed in the presence of RNase to eliminate the possibility of RNA bridging. (C) Input ³⁵S-labeled proteins. (D) GST binding assays were performed with in vitro-translated WT ICP27, GST-TAP, and TAP truncations. A schematic diagram of the TAP/NXF1 protein is shown.