Three Epstein-Barr virus (EBV)-determined nuclear antigens induced by the BamHI E region of EBV DNA

In our previous study (Takada et al., 1986a), we showed that the BamHI E fragment of Epstein-Barr virus (EBV) DNA induces a nuclear antigen that is detected by human antisera against EBV-determined nuclear antigen (EBNA), when transfected into baby hamster kidney (BHK) cells. The present study shows that the sub-fragment containing the central open reading frame BERF2b of the BamHI E fragment (Baer et al., 1984) is responsible for nuclear antigen induction. In addition, 2 fragments corresponding to 2 other open reading frames of the BamHI E, BERFI and BERF4 also induce nuclear antigens upon transfection into BHK cells. These 3 antigens, designated RF2b, RFI and RF4 antigens, were serologically classified as EBNA and antigenically distinct. In immunoblotting analysis of latently EBV-infected BJ-B95-8 cells, 3 high-molecular-weight polypeptides (136, 142 and 147 kDa) were identified by anti-EBNA sera. Immunoblotting analysis of transfected BHK cells indicated that the RF2b antigen is 145 kDa in its native form and antigenically related to the 147-kDa protein of BJ-B95-8 cells. Although RFI and RF4 antigens were not detected by immunoblotting, reactivities of sera with RFI and RF4 antigens in the immunofluorescence test were correlated with those of sera with the 136- and 142-kDa polypeptides of BJ-B95-8 cells, respectively. The results suggest that 3 high-molecular-weight proteins of latently EBV-infected cells are encoded by 3 open reading frames of the BamHI E DNA fragment.