Deletion of DNA encoding the first five transmembrane domains of Epstein-Barr virus latent membrane proteins 2A and 2B.
A recombinant Epstein-Barr virus (EBV) was constructed, with a positive-selection marker inserted at the site of a deletion of a DNA segment which encodes the first five transmembrane domains of LMP2A and LMP2B. Despite the mutation, the mutant recombinant EBV was able to initiate and maintain primary B-lymphocyte growth transformation in vitro. Cells transformed with the mutant recombinant were not different from wild-type virus transformants in initial or long-term outgrowth, sensitivity to limiting cell dilution, or serum requirement. Expression of EBNA1, EBNA2, EBNA3A, EBNA3C, and LMP1 and permissivity for lytic EBV infection were also unaffected by the LMP2 deletion mutation. These results complete the molecular genetic studies proving LMP2 is dispensable for primary B-lymphocyte growth transformation, latent infection, and lytic virus replication in vitro.
Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115.
R
R
Longnecker
C L
CL
Miller
B
B
Tomkinson
X Q
XQ
Miao
E
E
Kieff
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics; D001402:B-Lymphocytes / Q000382:microbiology; D001483:Base Sequence; D015139:Blotting, Southern; D002472:Cell Transformation, Viral; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D015183:Restriction Mapping; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014763:Viral Matrix Proteins / Q000096:biosynthesis* / Q000235:genetics; D014779:Virus Replication
D000956:Antigens, Viral; D004279:DNA, Viral; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D009838:Oligodeoxyribonucleotides; D014763:Viral Matrix Proteins