Marked, transient inhibition of expression of the Epstein-Barr virus latent membrane protein gene in Burkitt's lymphoma cell lines by electroporation.
The Raji, EB1, and EB2 cell lines are derived from Epstein-Barr virus (EBV)-positive Burkitt's lymphomas. EBV gene products associated with viral latency, including latent membrane protein (LMP) and Epstein-Barr nuclear antigen 2 (EBNA-2), are expressed in these cell lines. We have found that transfection of all three of these cell lines by electroporation in both the presence and the absence of foreign DNA resulted in a marked decrease in expression of the endogenous EBV gene encoding LMP. An analysis of this response in Raji cells revealed that the level of RNA of this gene was decreased transiently and returned to normal levels by 7 days after transfection. The level of LMP protein was also reduced after transfection. No difference in survival was detected in electroporated versus unperturbed Raji cells. The level of mRNA encoding a modulator of the LMP gene, EBNA-2, was unchanged by electroporation. However, the level of another EBV transcript, BHLF-I, was reduced. The effect of electroporation could not be attributed to flux of Ca2+, Na+, K+, or Cl- ions across the plasma membrane. Expression of LMP in several lymphoblastoid cell lines was unaffected by electroporation.
Gahn
T A
TA
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Sugden
B
B
D000838:Anions; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D002051:Burkitt Lymphoma / Q000382:microbiology*; D002414:Cations, Monovalent; D004268:DNA-Binding Proteins / Q000235:genetics; D018274:Electroporation; D019309:Epstein-Barr Virus Nuclear Antigens; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000235:genetics; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics
D000838:Anions; D000956:Antigens, Viral; D002414:Cations, Monovalent; D004268:DNA-Binding Proteins; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D019309:Epstein-Barr Virus Nuclear Antigens; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D014763:Viral Matrix Proteins; D015678:Viral Structural Proteins