Expression of a nuclear and a cytoplasmic Epstein-Barr virus early antigen after DNA transfer: cooperation of two distant parts of the genome for expression of the cytoplasmic antigen.

Expression of Epstein-Barr virus (EBV) antigens was studied after transfection of cloned EBV DNA fragments into baby hamster kidney (BHK) cells. A set of seven widely overlapping clones covering the whole genome of the non-defective Epstein-Barr virus strain M-ABA was used for transfection. Transfer of the cosmid clones into BHK cells resulted in expression of two distinct antigens, as revealed by indirect immunofluorescence using human anti-EBV sera. Staining with human sera of different reactivity against EBV-associated antigens revealed that both types of antigens were related to the early antigen complex. The first type of antigen was detected only in the nuclei of BHK cells that had received DNA of a clone containing HindIII-G, -H, -E, -I2, -O, -I1, and -P. The second type of antigen was found in the cytoplasm of cells cotransfected with clones containing Sal-A and HindIII-I2, -O, -I1, -P, and -C, whereas transfection of both individual clones failed to induce the antigen. Further analysis with subclones identified HindIII-G (5 kilobases) and HindIII-I2 (3 kilobases) plus the rightmost 3 kilobases of Sal-A as the sequences responsible for expression of the nuclear and the cytoplasmic antigen, respectively. The fact that two distant regions of the viral genome are required for expression of a viral antigen provides evidence for intergenomic regulation that can be studied in vitro.