Epstein-Barr virus nuclear antigen 2 (EBNA2)-oestrogen receptor fusion proteins complement the EBNA2-deficient Epstein-Barr virus strain P3HR1 in transformation of primary B cells but suppress growth of human B cell lymphoma lines.
To develop a transformation system with a conditional Epstein-Barr virus nuclear antigen 2 (EBNA2) gene, we fused the hormone binding domain of the oestrogen receptor to the N or C terminus of EBNA2. In promoter transactivation as well as primary B cell transformation assays these chimeric EBNA2 proteins are able to substitute for wild-type EBNA2 in the presence of oestrogen. Here we provide evidence that this transformation is the result of double infection of a cell with two virions, the P3HR1 virus genome and a mini-EBV plasmid carrying the chimeric EBNA2 gene. Unexpectedly, expression of the same EBNA2-oestrogen receptor fusion protein in established human B cell lymphoma lines resulted in growth retardation or growth arrest upon the addition of oestrogen. By titrating the oestrogen concentration in these stably transfected cells, the growth retarding and the transactivating function of the chimeric proteins could not be dissociated. We propose that growth inhibition of established B cell lymphoma lines is a novel function of EBNA2 which has not been detected in the absence of an inducible system. It remains open whether the growth retarding property of the EBNA2-oestrogen receptor fusion protein in B cell lymphoma lines is due to unphysiologically high expression of the chimeric protein or to interference with a cellular programme driving proliferation in these cell lines.
Kempkes
B
B
GSF-Forschungszentrum für Umwelt und Gesundheit, Institut für Molekularbiologie und Tumorgenetik, München, Germany.
Zimber-Strobl
U
U
Eissner
G
G
Pawlita
M
M
Falk
M
M
Hammerschmidt
W
W
Bornkamm
G W
GW
D000956:Antigens, Viral / Q000235:genetics / Q000502:physiology*; D001402:B-Lymphocytes / Q000502:physiology*; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D002472:Cell Transformation, Viral; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D019309:Epstein-Barr Virus Nuclear Antigens; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000473:pathology*; D008969:Molecular Sequence Data; D011960:Receptors, Estrogen / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D015533:Transcriptional Activation
D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins