Raji cells, a CR2-positive Burkitt lymphoma cell line, incubated in normal human serum, activate C3 and fix C3-derived fragments. The presence of these molecules on the cell surface does not affect subsequent Epstein-Barr virus (EBV) binding but it prevents superinfection. On the other hand, EBV superinfection is enhanced if Raji cells were incubated with heat-inactivated serum whose C3 fragments may bind only through receptor-binding sites. These results indicate that the region on cell surface offering the covalent site to C3 fragments would be essential for EBV superinfection. Incubation of Raji cells for 1 min with EBV results in the phosphorylation of CR2 and of a high-molecular-weight protein followed by their dephosphorylation, completed already after 20 min. This finding ascribes to EBV a prompt action through its receptor, different from that of other compounds causing a prolonged CR2 phosphorylation. Our data suggest that at least two binding sites are required for EBV superinfection of Raji cells or that specific patterns of CR2 phosphorylation may modulate Raji superinfection by EBV.