Transfer of the Epstein-Barr virus (EBV) DNA fragment coding for EBNA-1, the putative transforming antigen of EBV, into normal human lymphocytes: gene expression without cell transformation.
Fresh human lymphocytes were transfected with cloned EBV DNA fragments containing the coding exon for EBNA-1. DNA-loaded reconstituted Sendai Virus envelopes (RSVE) were used for efficient gene transfer. EBNA was detected by immunofluorescence in 1-5% of cells transfected with either the cloned BamHl K fragment of EBV DNA (5.1 kb) or the recombinant plasmid pSV3neoEBNA1, containing only the 2.0kb EBNA-1 coding exon. EBV-specific mRNA was detected by hybridization up to 14 days after DNA transfer. Quantitation of mRNA by laser densitometry revealed that the transcription level was similar to that obtained after infection with an intact virus. However, no effect on cellular proliferation was observed by (3H)-thymidine incorporation assays, and transformation was not achieved. We conclude that though EBNA-1 may be necessary for cellular immortalization by EBV, it alone is not sufficient.
D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics*; D001483:Base Sequence; D002472:Cell Transformation, Viral; D004247:DNA / Q000096:biosynthesis; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000502:physiology*; D015245:Deoxyribonuclease BamHI; D019309:Epstein-Barr Virus Nuclear Antigens; D005455:Fluorescent Antibody Technique; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D010222:Parainfluenza Virus 1, Human; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D014162:Transfection
D000956:Antigens, Viral; D004279:DNA, Viral; D019309:Epstein-Barr Virus Nuclear Antigens; D012333:RNA, Messenger; D012367:RNA, Viral; D004247:DNA; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI