A protein kinase activity in lymphocytic choriomeningitis virus and identification of the phosphorylated product using monoclonal antibody.

A cyclic AMP-independent protein kinase activity was found in purified preparations of the Armstrong CA 1371 strain of lymphocytic choriomeningitis virus (LCMV). Using the exquisite sensitivity of monoclonal antibodies to LCMV polypeptides, the internal nucleocapsid N protein was identified as the major virus-specific phosphorylated product of the endogenous protein kinase activity. This was accompanied by an increase in the electrophoretic mobility of N protein as detected by SDS-PAGE. After solubilization of the virus with 1% Nonidet P40 approximately 81% of the endogenous protein kinase activity remained associated with LCMV nucleocapsids recovered by equilibrium centrifugation at a density of 1.25 g/cm-3 in a linear renograffin gradient. Specific phosphorylation of N protein was reconfirmed in the purified nucleocapsid fraction and both phosphoserine and phosphothreonine found to be the phosphorylated products of the kinase reaction. Although the significance of this enzyme remains unclear, the presence of a protein kinase within LCMV may allow the regulation of LCMV replication and maturation by phosphorylation of virus-specific polypeptides. These events may in turn play a key role in determining the nature and outcome of LCMV infection.