FIG. 6.

Changing Z residue 186 from serine to threonine affects the ability of Z to induce lytic EBV infection in a methylation-dependent manner. (A) 293/EBV-ZKO cells (20) were treated with 5-aza-2′-dc or not treated, as shown, prior to transfection with either empty vector (Vector) or a vector expressing wild-type Z (Wt Z) or Z(S186T). The methylation status of the EBV genome after the 5-aza-2′-dc treatment is indicated. Immunoblots were performed on cell extracts 2 days after transfection with monoclonal antibody directed against Z, BMRF1 (an early lytic EBV protein), R, or β-actin, as indicated. (B) 293/EBV-ZKO cells pretreated with 5-aza-2′-dc were transfected with empty vector or a vector expressing R, Z(S186T), or both R and Z(S186T). The methylation status of the EBV genome is indicated. Immunoblotting was performed for Z, BMRF1, or β-actin, as indicated. +, present; −, absent.