Biochemical characterization of two Epstein-Barr virus early antigen-associated phosphopolypeptides

Epstein-Barr virus (EBV) nonproducer cells NC37 induced to viral early antigen (EA) synthesis by the tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA) were labeled at Day 4 after induction with 32P, and were analyzed by immunoprecipitation with human EA-positive sera. By employing this method the appearance of two phosphopolypeptides of 50 and 58K (pp50 and pp58) was well correlated with EA complex. Partial V8 protease digestion and two-dimensional peptide analysis revealed that the polypeptides pp50 and pp58 are related. The analysis of phosphoamino acids indicated that pp58 contains phosphoserine and phosphothreonine to the same percentage, whereas in pp50 only phosphoserine was found. The analysis of the subcellular distribution revealed that pp50 and pp58 are located in the chromatin. Both phosphopolypeptides exhibit DNA-binding activity, and are recognized by two monoclonal antibodies described recently (R3 and 1108-1).