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.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA042245-03
Application #
3183263
Study Section
Virology Study Section (VR)
Project Start
1986-04-01
Project End
1989-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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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

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