Human p32: a coactivator for Epstein-Barr virus nuclear antigen-1-mediated transcriptional activation and possible role in viral latent cycle DNA replication.
The Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) is required for the maintenance of the viral chromosome in latently infected, proliferating cells and plays a role in latent cycle DNA replication. EBNA-1 also functions as a positive and negative regulator of EBV gene expression. We have investigated the interaction of EBNA-1 with p32, a host mitochondrial protein that associates with EBNA-1 in EBV-positive Burkitt's lymphoma cells. Using a chromatin immunoprecipitation assay, we found that a fraction of p32 localizes to the viral latent cycle origin of DNA replication oriP in vivo. p32 binds EBNA-1 independently of other proteins or DNA. EBNA-1 variants lacking one of two p32 binding elements did not interact stably with p32 in cultured cells and were defective for both transcriptional activation of a reporter gene linked to oriP FR and replication and/or maintenance of a plasmid bearing oriP. These results support a role for p32 in transcriptional activation by EBNA-1 and suggest that p32 plays a role in EBV latent cycle DNA replication.
Van Scoy
S
S
Department of Molecular Genetics and Microbiology, State University of New York, Stony Brook, New York 11794, USA.
Watakabe
I
I
Krainer
A R
AR
Hearing
J
J
D000818:Animals; D001665:Binding Sites; D002051:Burkitt Lymphoma / Q000378:metabolism / Q000473:pathology; D002352:Carrier Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000502:physiology; D006801:Humans; D018960:Hyaluronan Receptors; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011233:Precipitin Tests; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D018741:Replication Origin / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics; D013379:Substrate Specificity; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured; D017735:Virus Latency / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
C087299:C1QBP protein, human; D002352:Carrier Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D018960:Hyaluronan Receptors; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D011506:Proteins; D004247:DNA