The only domain which distinguishes Epstein-Barr virus latent membrane protein 2A (LMP2A) from LMP2B is dispensable for lymphocyte infection and growth transformation in vitro; LMP2A is therefore nonessential.
Using second-site homologous recombination, Epstein-Barr virus (EBV) recombinants were constructed which carry an LMP2A mutation terminating translation at codon 19. Despite the absence of LMP2A or LMP2A cross-reactive protein, the recombinants were able to initiate and maintain primary B-lymphocyte growth transformation in vitro. EBNA1, EBNA2, and LMP1 expression was unaffected by the LMP2A mutation. The LMP2A mutant recombinant EBV-infected lymphoblastoid cell lines (LCLs) were identical to wild-type recombinant EBV-infected control LCLs with respect to initial outgrowth, subsequent growth, sensitivity to limiting cell dilution, sensitivity to low serum, and growth in soft agarose. The permissivity of LCLs for lytic EBV infection and virus replication was also unaffected by the LMP2A mutation.
Longnecker
R
R
Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115.
Miller
C L
CL
Miao
X Q
XQ
Marchini
A
A
Kieff
E
E
D000956:Antigens, Viral / Q000235:genetics; D001402:B-Lymphocytes / Q000382:microbiology*; D001483:Base Sequence; D002051:Burkitt Lymphoma; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D003062:Codon; D003429:Cross Reactions; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D011995:Recombination, Genetic; D014763:Viral Matrix Proteins / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication
D000956:Antigens, Viral; D003062:Codon; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D008565:Membrane Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins