Expression of the Epstein-Barr virus encoded EBNA-1 gene in stably transfected human and murine cell lines.
Five murine and 3 human tumor cell lines were transfected with a retroviral vector that carries the EBV encoded EBNA-1 gene. All cell lines expressed intranuclear EBNA-1 as detected by anticomplement immunofluorescence and Western blot assays. The cell lines differed in the level of EBNA-1 expression and the size of the protein. The internal major late promoter of adenovirus was efficient in directing the transcription of EBNA-1 in the human lymphoma line BJAB, the murine T-cell lymphoma Tikaut, RBL-5, EL-4 and in the mouse sarcoma line MSWBS but was less efficient in Ramos, an EBV negative Burkitt lymphoma line, the human T-cell leukemia line 1301TK and the P815-X2 mouse mastocytoma line. All transfected lines except MSWBS contained EBNA-1 in a truncated form. The truncated EBNA-1 polypeptide reacted with the conventional human antibody reagents in an EBNA specific fashion but failed to bind rabbit or human antibody directed against the glycine-alanine repeat sequence. MSWBS contained a truncated as well as a full size EBNA-1 polypeptide. It also reacted with antibody directed against the glycine-alanine repeat. This indicates that the repeat sequence is regularly affected by the truncation.
Patel
G V
GV
Department of Tumor Biology, Karolinska Institutet, Stockholm, Sweden.
Masucci
M G
MG
Winberg
G
G
Klein
G
G
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics*; D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens; D015321:Gene Rearrangement; D004854:Herpesvirus 4, Human / Q000302:isolation & purification; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection
D000956:Antigens, Viral; D019309:Epstein-Barr Virus Nuclear Antigens