Secretion of Epstein-Barr virus-encoded BARF1 oncoprotein from latently infected B cells.
Epstein-Barr virus (EBV) encodes two oncogenes, LMP1(Latent Membrane Protein-1) and BARF1 (BamH1-A Reading Frame-1). LMP1 belongs to latent gene family and BARF1 is considered so far as one of early gene family. However BARF1 oncogene was expressed highly in Nasopharyngeal (NPC) and gastric (GC) carcinoma as a type II latency, and in EBV-positive Akata cell and primary epithelial cell infected in vitro by EBV as type I latency. Its expression was also reported in Burkitt's lymphoma's biopsy frequent in Malawi in Africa as well as in nasal NK/T-cell lymphoma. We recently observed a massive secretion of BARF1 protein in serum and saliva of NPC patients. NPC-derived c666-1 epithelial cells also expressed and secreted BARF1 protein without other lytic genes expression. We asked whether this oncogene belongs to latent gene family. To investigate, we examined its transcriptional and translational expression in IB4 and Akata B cells where both cell lines belong to latent cell family. Transcriptional expression was analyzed by RT-PCR. As BARF1 protein is one of secreted proteins, its translational expression was analyzed by immunoblot after concentration of culture medium. Secreted BARF1 protein was futher purified by concanavalin A affinity column. BARF1 was transcribed in both EBV-positive AKATA and IB4 cells, and BARF1 protein was secreted from these latently infected human B cells. Its secretion does not depend EBV genome form in infected cells. Both episomal and integrated form of EBV genome were capable of expressing BARF1 gene. These results suggests that BARF1 is expressed in latent stage and increases its expression during lytic stage.
Laboratoire de Virologie Moléculaire, Virologie et Pathogenèse Humaine, FRE 3011, CNRS, Faculté de Médecine Laennec, Université Claude Bernard Lyon-1, Rue Guillaume Paradin, 69372, Lyon Cedex 08, France. sylvie.fiorini@sante.univ-lyon1.fr
Sylvie
S
Fiorini
Tadamasa
T
Ooka
D001402:B-Lymphocytes / Q000821:virology*; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D003470:Culture Media / Q000737:chemistry; D020869:Gene Expression Profiling; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D008295:Malawi; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
C086495:BARF1 protein, Human herpesvirus 4; D003470:Culture Media; D012333:RNA, Messenger; D014764:Viral Proteins