Epstein-Barr virus latent membrane protein 2B (LMP2B) co-localizes with LMP2A in perinuclear regions in transiently transfected cells.
Epstein-Barr virus is a human gammaherpesvirus that infects and establishes latency in B lymphocytes in vivo. The latent membrane protein 2 (LMP2) gene is expressed in latently infected B cells and encodes two protein isoforms, LMP2A and LMP2B, that are identical except for an additional N-terminal 119 aa cytoplasmic domain which is present in the LMP2A isoform. A panel of fusion proteins was constructed in which the fluorescent enhanced green fluorescent protein and DsRed protein domains were fused to the N- and C-termini of LMP2A and LMP2B. By fluorescence microscopy, LMP2B localized to perinuclear regions of both live and fixed transiently transfected cells. Co-localization was detected with markers for the endoplasmic reticulum and the trans-Golgi network. No evidence of co-localization of LMP2B with endosomes or surface expression was obtained. Transiently expressed LMP2B co-localized with transiently or constitutively expressed LMP2A. Confocal microscopy confirmed that LMP2A proteins localized to intracellular perinuclear compartments with markers for the trans-Golgi network. Only LMP2A proteins with C-terminal truncations were detected in the plasma membrane with extracellular loop1 epitope tags. These results indicate that the transmembrane domain of LMP2 proteins possess intracellular retention signals and suggest that LMP2A-mediated signalling effects are likely to be ectopic, originating from sites inside the cell close to the nucleus.
Lynch
David T
DT
Department of Infectious Diseases and Microbiology, Graduate School of Public Health, 130 Desoto Street, Pittsburgh, PA 15213, USA1.
Zimmerman
Jeffrey S
JS
Department of Infectious Diseases and Microbiology, Graduate School of Public Health, 130 Desoto Street, Pittsburgh, PA 15213, USA1.
Rowe
David T
DT
Department of Infectious Diseases and Microbiology, Graduate School of Public Health, 130 Desoto Street, Pittsburgh, PA 15213, USA1.
D018941:Antigens, CD19 / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry*; D006801:Humans; D018613:Microscopy, Confocal; D011993:Recombinant Fusion Proteins / Q000032:analysis; D014162:Transfection; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000032:analysis*
D018941:Antigens, CD19; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins