scholarly journals Epstein-Barr virus nuclear antigen forms a complex that binds with high concentration dependence to a single DNA-binding site.

1987 ◽  
Vol 61 (2) ◽  
pp. 465-471 ◽  
Author(s):  
G Milman ◽  
E S Hwang
Keyword(s):  
Dna Binding ◽  
Binding Site ◽  
Concentration Dependence ◽  
Epstein Barr Virus ◽  
Nuclear Antigen ◽  
High Concentration ◽  
Barr Virus ◽  
Dna Binding Site ◽  
Epstein Barr

scholarly journals Epstein-Barr Virus Nuclear Antigen 3C Activates the Latent Membrane Protein 1 Promoter in the Presence of Epstein-Barr Virus Nuclear Antigen 2 through Sequences Encompassing an Spi-1/Spi-B Binding Site

2000 ◽  
Vol 74 (11) ◽  
pp. 5151-5160 ◽  
Author(s):  
Bo Zhao ◽  
Clare E. Sample
Keyword(s):  
Dna Binding ◽  
Binding Site ◽  
Binding Sites ◽  
Epstein Barr Virus ◽  
Latent Membrane Protein ◽  
Nuclear Antigen ◽  
Latent Membrane Protein 1 ◽  
Barr Virus ◽  
Epstein Barr

ABSTRACT The Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA-3C) protein is a transcriptional regulator of viral and cellular genes that is essential for EBV-mediated immortalization of B lymphocytes in vitro. EBNA-3C can inhibit transcription through an association with the cellular DNA-binding protein Jκ, a function shared by EBNA-3A and EBNA-3B. Here, we report a mechanism by which EBNA-3C can activate transcription from the EBV latent membrane protein 1 (LMP-1) promoter in conjunction with EBNA-2. Jκ DNA-binding sites were not required for this activation, and a mutant EBNA-3C protein unable to bind Jκ activated transcription as efficiently as wild-type EBNA-3C, indicating that EBNA-3C can regulate transcription through a mechanism that is independent of Jκ. Furthermore, activation of the LMP-1 promoter is a unique function of EBNA-3C, not shared by EBNA-3A and EBNA-3B. The DNA element through which EBNA-3C activates the LMP-1 promoter includes a Spi-1/Spi-B binding site, previously characterized as an important EBNA-2 response element. Although this element has considerable homology to mouse immunoglobulin light chain promoter sequences to which the mouse homologue of Spi-1 binds with its dimerization partner IRF4, we demonstrate that the IRF4-like binding sites in the LMP-1 promoter do not play a role in EBNA-3C-mediated activation. Both EBNA-2 and EBNA-3C were required for transcription mediated through a 41-bp region of the LMP-1 promoter encompassing the Spi binding site. However, EBNA-3C had no effect on transcription mediated in conjunction with the EBNA-2 activation domain fused to the GAL4 DNA-binding domain, suggesting that it does not function as an adapter between EBNA-2 and the cellular transcriptional machinery. Like EBNA-2, EBNA-3C bound directly to both Spi-1 and Spi-B in vitro. This interaction was mediated by a region of EBNA-3C encompassing a likely basic leucine zipper (bZIP) domain and the ets domain of Spi-1 or Spi-B, reminiscent of interactions between bZIP and ets domains of other transcription factors that result in their targeting to DNA. There are many examples of regulation of the hematopoietic-specific Spi transcription factors through protein-protein interactions, and a similar regulation by EBNA-3C, in conjunction with EBNA-2, is likely to be an important and unique contribution of EBNA-3C to EBV-mediated immortalization.

scholarly journals Development of a High-Throughput Screen for Inhibitors of Epstein-Barr Virus EBNA1

2010 ◽  
Vol 15 (9) ◽  
pp. 1107-1115 ◽  
Author(s):  
Scott Thompson ◽  
Troy Messick ◽  
David C. Schultz ◽  
Melvin Reichman ◽  
Paul M. Lieberman
Keyword(s):  
Dna Binding ◽  
Binding Site ◽  
Small Molecule ◽  
High Throughput ◽  
Epstein Barr Virus ◽  
Latent Infection ◽  
Dna Binding Protein ◽  
Barr Virus ◽  
Dna Binding Site ◽  
Epstein Barr

Latent infection with Epstein-Barr virus (EBV) is a carcinogenic cofactor in several lymphoid and epithelial cell malignancies. At present, there are no small-molecule inhibitors that specifically target EBV latent infection or latency-associated oncoproteins. EBNA1 is an EBV-encoded sequence-specific DNA binding protein that is consistently expressed in EBV-associated tumors and required for stable maintenance of the viral genome in proliferating cells. EBNA1 is also thought to provide cell survival function in latently infected cells. In this work, the authors describe the development of a biochemical high-throughput screening (HTS) method using a homogeneous fluorescence polarization (FP) assay monitoring EBNA1 binding to its cognate DNA binding site. An FP-based counterscreen was developed using another EBV-encoded DNA binding protein, Zta, and its cognate DNA binding site. The authors demonstrate that EBNA1 binding to a fluorescent-labeled DNA probe provides a robust assay with a Z factor consistently greater than 0.6. A pilot screen of a small-molecule library of ~14,000 compounds identified 3 structurally related molecules that selectively inhibit EBNA1 but not Zta. All 3 compounds had activity in a cell-based assay specific for the disruption of EBNA1 transcription repression function. One of the compounds was effective in reducing EBV genome copy number in Raji Burkitt lymphoma cells. These experiments provide a proof of concept that small-molecule inhibitors of EBNA1 can be identified by biochemical HTS of compound libraries. Further screening in conjunction with medicinal chemistry optimization may provide a selective inhibitor of EBNA1 and EBV latent infection.

scholarly journals High-resolution footprints of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1.

1989 ◽  
Vol 9 (6) ◽  
pp. 2738-2742 ◽  
Author(s):  
A S Kimball ◽  
G Milman ◽  
T D Tullius
Keyword(s):  
Dna Binding ◽  
Epstein Barr Virus ◽  
Binding Domain ◽  
Nuclear Antigen ◽  
Dna Binding Domain ◽  
Major Groove ◽  
Barr Virus ◽  
Dna Binding Site ◽  
Epstein Barr ◽  
Hydroxyl Radical Footprinting

The DNA-binding domain of Epstein-Barr virus nuclear antigen 1 was found by hydroxyl radical footprinting to protect backbone positions on one side of its DNA-binding site. The guanines contacted in the major groove by the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 were identified by methylation protection. No difference was found in the interaction of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 with tandemly repeated and overlapping binding sites.

High-resolution footprints of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1

1989 ◽  
Vol 9 (6) ◽  
pp. 2738-2742
Author(s):  
A S Kimball ◽  
G Milman ◽  
T D Tullius
Keyword(s):  
Dna Binding ◽  
Epstein Barr Virus ◽  
Binding Domain ◽  
Nuclear Antigen ◽  
Dna Binding Domain ◽  
Major Groove ◽  
Barr Virus ◽  
Dna Binding Site ◽  
Epstein Barr ◽  
Hydroxyl Radical Footprinting

The DNA-binding domain of Epstein-Barr virus nuclear antigen 1 was found by hydroxyl radical footprinting to protect backbone positions on one side of its DNA-binding site. The guanines contacted in the major groove by the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 were identified by methylation protection. No difference was found in the interaction of the DNA-binding domain of Epstein-Barr virus nuclear antigen 1 with tandemly repeated and overlapping binding sites.

scholarly journals Characterization of the DNA-binding site repertoire for the Epstein - Barr virus transcription factor R

1994 ◽  
Vol 22 (7) ◽  
pp. 1172-1178 ◽  
Author(s):  
Henri Gruffat ◽  
Alain Sergeant
Keyword(s):  
Transcription Factor ◽  
Dna Binding ◽  
Binding Site ◽  
Epstein Barr Virus ◽  
Barr Virus ◽  
Virus Transcription ◽  
Dna Binding Site ◽  
Epstein Barr

scholarly journals Mutational analysis of the Epstein--Barr virus nuclear antigen 2 by far-Western blotting and DNA-binding studies

1996 ◽  
Vol 77 (5) ◽  
pp. 991-996 ◽  
Author(s):  
C. Sauder ◽  
N. Gotzinger ◽  
W. H. Schubach ◽  
G. C. Horvath ◽  
E. Kremmer ◽  
...  
Keyword(s):  
Dna Binding ◽  
Western Blotting ◽  
Epstein Barr Virus ◽  
Mutational Analysis ◽  
Nuclear Antigen ◽  
Binding Studies ◽  
Barr Virus ◽  
Dna Binding Studies ◽  
Epstein Barr

scholarly journals Overexpression, Purification, and Crystallization of the DNA Binding and Dimerization Domains of the Epstein-Barr Virus Nuclear Antigen 1

1995 ◽  
Vol 270 (35) ◽  
pp. 20556-20559 ◽  
Author(s):  
Jean A. Barwell ◽  
Alexey Bochkarev ◽  
Richard A. Pfuetzner ◽  
Harry Tong ◽  
Daniel S. C. Yang ◽  
...  
Keyword(s):  
Dna Binding ◽  
Epstein Barr Virus ◽  
Nuclear Antigen ◽  
Barr Virus ◽  
Epstein Barr

scholarly journals Small molecule inhibition of Epstein–Barr virus nuclear antigen-1 DNA binding activity interferes with replication and persistence of the viral genome

2014 ◽  
Vol 104 ◽  
pp. 73-83 ◽  
Author(s):  
Eun Kyung Lee ◽  
Sun Young Kim ◽  
Ka-Won Noh ◽  
Eun Hye Joo ◽  
Bo Zhao ◽  
...  
Keyword(s):  
Dna Binding ◽  
Small Molecule ◽  
Viral Genome ◽  
Epstein Barr Virus ◽  
Binding Activity ◽  
Nuclear Antigen ◽  
Barr Virus ◽  
Dna Binding Activity ◽  
Small Molecule Inhibition ◽  
Epstein Barr

Sequence-specific DNA binding of the Epstein-Barr virus nuclear antigen (EBNA-1) to clustered sites in the plasmid maintenance region

Cell ◽  
1985 ◽  
Vol 42 (3) ◽  
pp. 859-868 ◽  
Author(s):  
Dan R. Rawlins ◽  
Gregory Milman ◽  
S.Diane Hayward ◽  
Gary S. Hayward
Keyword(s):  
Dna Binding ◽  
Epstein Barr Virus ◽  
Nuclear Antigen ◽  
Plasmid Maintenance ◽  
Barr Virus ◽  
Epstein Barr
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