FIG 4.

Identification of ORF44p as the binding partner of ORF49p by proteomic analysis and their in vitro binding assay. (A) pOka-infected MeWo cells expanded by cell-to-cell spread with full CPE at 2 to 3 days postinfection were lysed with RIPA buffer, and the binding molecules were coimmunoprecipitated with ORF49p using an anti-ORF49 Ab (lane 1). Mock-infected MeWo cells were used as a negative control (lane 2). The immunoprecipitates (ip) were electrophoretically separated and visualized by silver staining. (B) ORF49p expressed in and purified from MeWo cells was incubated with purified GST-ORF44 (lane 1), GST-ORF44F129A (lane 2), or GST-ORF44P (lane 3). Bound proteins were electrophoretically separated and visualized by anti-GST Abs (upper panel) and anti-ORF49 Abs (lower panel).