Epstein-Barr Virus (EBV) causes infectious mononucleosis and is carried as a latent infection by 80% of the adult American population. Individuals who are immunologically compromised, such as AIDS victims, and patient undergoing organ or bone-marrow transplantation, frequently develop symptoms of re-activated EBV infection. Incidences of EBV-positive lymphomas in these patients has also been a cause for concern. An understanding of the factors involved in viral replication and maintance of latency would, in the long term, assist in the management of these patients. EBV is also believed to be a causative factor in Africa Burkitt lymphoma and nasopharyngeal carcinoma (which occurs pedominatly in Chinese ethnic groups) since the cells of these tumors contain EBV DNA and express the EBNA-1 antigen. The long range goals fo the proposed research is to define the molecular mechanisms involved in the replication of Epstein-Barr Virus DNA and in the maintenance of viral latency in transformed cells.
The specific aims of this application are to investigate the biological and biochemical processes involving the EBV nuclear antigen EBNA-1 and the regions of EBV DNA with which it interacts and will include: 1. Isolation of the intact EBNA-1 protein. 2. Examination of the parameters of binding of EBNA-1 to its DNA binding sites. 3. Examination of the biological implications of manipulating the numbers and affinities of the EBNA-1 binding sites in ori-P. 4. Determination of the minimal region of EBNA-1 required for specific DNA binding, plasmid maintenance, nuclear localization of EBNA-1, and chromosomal association of EBNA-1. 5. Invesitgation of a possible role for EBNA-1 in regulation of biral gene expression, and 6. Identification of lytic EBV orgins of replication. Specific reagents will be generated using recombinant Dna thechnology and assay procedures will include DNAaseI footprinting, nitrocellulose filterbinding assays, DNA transfection, CAT assays and immunoflorescene.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
Project #
Application #
Study Section
Virology Study Section (VR)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Johns Hopkins University
Schools of Medicine
United States
Zip Code
Lo, Angela Kwok Fung; To, Ka Fai; Lo, Kwok Wai et al. (2007) Modulation of LMP1 protein expression by EBV-encoded microRNAs. Proc Natl Acad Sci U S A 104:16164-9
Lee, Jae Myun; Lee, Kyoung-Ho; Weidner, Magdalena et al. (2002) Epstein-Barr virus EBNA2 blocks Nur77- mediated apoptosis. Proc Natl Acad Sci U S A 99:11878-83
Zhou, S; Hayward, S D (2001) Nuclear localization of CBF1 is regulated by interactions with the SMRT corepressor complex. Mol Cell Biol 21:6222-32
Zhang, J; Chen, H; Weinmaster, G et al. (2001) Epstein-Barr virus BamHi-a rightward transcript-encoded RPMS protein interacts with the CBF1-associated corepressor CIR to negatively regulate the activity of EBNA2 and NotchIC. J Virol 75:2946-56
Zhou, S; Fujimuro, M; Hsieh, J J et al. (2000) SKIP, a CBF1-associated protein, interacts with the ankyrin repeat domain of NotchIC To facilitate NotchIC function. Mol Cell Biol 20:2400-10
Zhou, S; Fujimuro, M; Hsieh, J J et al. (2000) A role for SKIP in EBNA2 activation of CBF1-repressed promoters. J Virol 74:1939-47
Smith, P R; de Jesus, O; Turner, D et al. (2000) Structure and coding content of CST (BART) family RNAs of Epstein-Barr virus. J Virol 74:3082-92
Chen, H; Smith, P; Ambinder, R F et al. (1999) Expression of Epstein-Barr virus BamHI-A rightward transcripts in latently infected B cells from peripheral blood. Blood 93:3026-32
Hsieh, J J; Zhou, S; Chen, L et al. (1999) CIR, a corepressor linking the DNA binding factor CBF1 to the histone deacetylase complex. Proc Natl Acad Sci U S A 96:23-8
Hsieh, J J; Henkel, T; Salmon, P et al. (1996) Truncated mammalian Notch1 activates CBF1/RBPJk-repressed genes by a mechanism resembling that of Epstein-Barr virus EBNA2. Mol Cell Biol 16:952-9

Showing the most recent 10 out of 22 publications