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PMID: 8445720 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Epstein-Barr virus nuclear proteins EBNA-3A and EBNA-3C are essential for B-lymphocyte growth transformation.

Journal of virology ·Vol. 67 ·No. 4 ·1993-04-00 ·Pages 2014-25

Tomkinson B, Robertson E, Kieff E

Abstract

Recombinant Epstein-Barr viruses (EBV) with a translation termination codon mutation inserted into the nuclear protein 3A (EBNA-3A) or 3C (EBNA-3C) open reading frame were generated by second-site homologous recombination. These mutant viruses were used to infect primary B lymphocytes to assess the requirement of EBNA-3A or -3C for growth transformation. The frequency of obtaining transformants infected with a wild-type EBNA-3A recombinant EBV was 10 to 15%. In contrast, the frequency of obtaining transformants infected with a mutant EBNA-3A recombinant EBV was only 1.4% (9 mutants in 627 transformants analyzed). Transformants infected with mutant EBNA-3A recombinant virus could be obtained only by coinfection with another transformation-defective EBV which provided wild-type EBNA-3A in trans. Cells infected with mutant EBNA-3A recombinant virus lost the EBNA-3A mutation with expansion of the culture. The decreased frequency of recovery of the EBNA-3A mutation, the requirement for transformation-defective EBV coinfection, and the inability to maintain the EBNA-3A mutation indicate that EBNA-3A is essential or critical for lymphocyte growth transformation and that the EBNA-3A mutation has a partial dominant negative effect. Five transformants infected with mutant EBNA-3C recombinant virus EBV were also identified and expanded. All five also required wild-type EBNA-3C in trans. Serial passage of the mutant recombinant virus into primary B lymphocytes resulted in transformants only when wild-type EBNA-3C was provided in trans by coinfection with a transformation-defective EBV carrying a wild-type EBNA-3C gene. A secondary recombinant virus in which the mutated EBNA-3C gene was replaced by wild-type EBNA-3C was able to transform B lymphocytes. Thus, EBNA-3C is also essential or critical for primary B-lymphocyte growth transformation.

MeSH Terms
Antigens, Viral/physiology B-Lymphocytes/microbiology Base Sequence Cell Transformation, Viral Cells, Cultured DNA-Binding Proteins/physiology Epstein-Barr Virus Nuclear Antigens Humans In Vitro Techniques Molecular Sequence Data Oligodeoxyribonucleotides/chemistry Polymerase Chain Reaction Restriction Mapping
Chemicals
Antigens, Viral DNA-Binding Proteins Epstein-Barr Virus Nuclear Antigens Oligodeoxyribonucleotides
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tomkinson B
Department of Microbiology and Molecular Genetics and Medicine, Harvard University, Boston, Massachusetts 02115.
Robertson E
Kieff E
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.4.2014-2025.1993
Published
1993-04-00
Pages
2014-25
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240270
Subset
IM
Grants
NCI NIH HHS · CA47006 · United States
External Links
PubMed source DOI: 10.1128/JVI.67.4.2014-2025.1993 PMC Full Text View journal details
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