Abstract

We have proposed to identify restriction fragments of EBV DNA that contain the origin of lytic viral DNA replication or of latent viral DNA replication. D98/HRI cells that carry P3HRI EBV genomes and grow in monolayer will be induced for viral DNA replication by IUDR and transfected with bacterial plasmid containing Amp(r), TK and viral DNA fragment but lacking sequence inhibitory to mammalian DNA replication. The plasmid with EBV fragment that can replicate together with P3HR viral DNA will be identified. D98/Raji TK- cells will be transfected with the same plasmid DNA with EBV DNA fragmetn and the fragments that can persist in D98/Raji cells will be identified. The region for the origin of viral DNA replication will be reduced to minimum necessary size by restriction enzymes, Bal 31 and/or DNAase 1 pratial digestion. If the plasmid containing the origin of latent viral DNA replication cannot persist in D98 cells that do not carry viral genome, then viral DNA fragments that provide functions for the plasmid DNA to persist in D98 cells will be identified. Finally celular DNA sequences hybridizable to the origin of latent viral DNA replication will be examined, based on an assumption that latent viral plasmid DNA and cellular DNA might share similar sequence for the origin of DNA replication.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA036895-02
Application #
3174505
Study Section
Experimental Virology Study Section (EVR)
Project Start
1984-12-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Tampa Bay Research Institute
Department
Type
DUNS #
City
Saint Petersburg
State
FL
Country
United States
Zip Code
33716

  Publications

Nonoyama, M; Wen, L T; Tanaka, A et al. (1990) Detection of 12-o-tetradecanoylphorbol-13-acetate-induced cellular proteins that compete with the Epstein-Barr virus nuclear antigen 1 (EBNA-1) for binding to a site within the Epstein-Barr virus oriP. Adv Exp Med Biol 278:125-36
Wen, L T; Lai, P K; Bradley, G et al. (1990) Interaction of Epstein-Barr viral (EBV) origin of replication (oriP) with EBNA-1 and cellular anti-EBNA-1 proteins. Virology 178:293-6
Wen, L T; Tanaka, A; Nonoyama, M (1989) Induction of anti-EBNA-1 protein by 12-O-tetradecanoylphorbol-13-acetate treatment of human lymphoblastoid cells. J Virol 63:3315-22
Bradley, G; Lancz, G; Tanaka, A et al. (1989) Loss of Marek's disease virus tumorigenicity is associated with truncation of RNAs transcribed within BamHI-H. J Virol 63:4129-35
Bradley, G; Hayashi, M; Lancz, G et al. (1989) Structure of the Marek's disease virus BamHI-H gene family: genes of putative importance for tumor induction. J Virol 63:2534-42
Yamamoto, M; Tabata, T; Smith, M et al. (1989) Cycloheximide-resistant gene of Epstein-Barr virus in freshly infected B lymphocytes. Virology 170:307-10
Sawada, K; Yamamoto, M; Tabata, T et al. (1989) Expression of EBNA-3 family in fresh B lymphocytes infected with Epstein-Barr virus. Virology 168:22-30
Wen, L T; Tanaka, A; Nonoyama, M (1988) Identification of Marek's disease virus nuclear antigen in latently infected lymphoblastoid cells. J Virol 62:3764-71
Nonoyama, M; Wen, L T; Tabata, T et al. (1988) A novel EBNA-1 titration method and putative anti-EBNA-1 protein. J Virol Methods 21:161-70
Harada, H; Sawada, K; Kudo, S et al. (1987) Development of cell systems to study viral gene transcription at the initial phase of Epstein-Barr virus infection. Virus Genes 1:73-82

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