RNAs induced by Epstein-Barr virus nuclear antigen 2 in lymphoblastoid cell lines.
Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA2) association with RBP-Jkappa is essential for regulation of virus and cell gene transcription and B lymphocyte transformation into infinitely proliferating lymphoblastoid cells (LCLs). To identify EBNA2-regulated cell genes in LCLs, an EBV recombinant that expresses EBNA2 with its C terminus fused in frame to a 4-hydroxytamoxifen (4HT)-dependent mutant estrogen receptor (E2HTF) was used to transform primary B lymphocytes to LCLs. In the presence of 4HT, E2HTF expression level and effects on the LMP1 promoter in transfected BJAB lymphoblasts were similar to EBNA2. In 4HT-supplemented medium, E2HTF EBV recombinant infected LCLs were also similar to EBNA2 LCLs in outgrowth but required higher serum and a restricted range of cell concentrations for consistent growth. In medium without 4HT, E2HTF localized to the cytoplasm, c-myc levels substantially decreased within 6 h, cells stopped growing, and levels of other EBNAs and LMP1 remained stable for 24 h. Over this 24-h period, 30 cell RNAs decreased 2-fold, and 51 other RNAs decreased 1.5-fold. These RNAs encode proteins important in cell adhesion or signaling, transcription, RNA processing, cell-cycle regulation, and survival. Real-time RT-PCR confirmed EBNA2-dependent expression of eight RNAs.
Zhao
Bo
B
Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
Maruo
Seiji
S
Cooper
Andrew
A
R Chase
Michael
M
Johannsen
Eric
E
Kieff
Elliott
E
Cahir-McFarland
Ellen
E
D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003201:Computers; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000473:pathology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis* / Q000235:genetics*; D051881:Receptor, Notch1 / Q000378:metabolism; D017455:Receptors, IgE / Q000378:metabolism; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology
D019309:Epstein-Barr Virus Nuclear Antigens; D016271:Proto-Oncogene Proteins c-myc; D051881:Receptor, Notch1; D017455:Receptors, IgE; D013629:Tamoxifen; C016601:afimoxifene; D012313:RNA