FIG. 1.

Interaction of ICP0 with members of the Src kinase family. (A) SH3 domains of the Src family protein kinases pull down ICP0. Equal amounts of infected-cell lysates were reacted for 4 h at 4°C with GST-tagged SH3 domains of SFK members Fgr, Src, neuronal Src, Fyn, and Yes. The proteins adhering to the glutathione-agarose beads were solubilized, electrophoretically separated in denaturing gels, and reacted with anti-ICP0 monoclonal antibody (top) or stained with Coomassie bright blue to show the GST-SH3 chimeric proteins used in each pull-down assay. (B) Src interacts only with the polypeptide comprising residues 245 to 510 of ICP0. GST-ICP0 chimeric proteins bound to agarose beads as described in Materials and Methods were reacted with equal amounts of uninfected HEK293 cell lysates. The proteins bound to the beads were solubilized, separated in denaturing gels, transferred to a nitrocellulose membrane, and reacted with antibodies against CIN85 (top), Src (middle), or GST (bottom). The lower gel shows the amounts of chimeric proteins loaded on the beads.