Repeat arrays in cellular DNA related to the Epstein-Barr virus IR3 repeat

1985 ◽  
Vol 5 (3) ◽  
pp. 457-465
Author(s):  
M Heller ◽  
E Flemington ◽  
E Kieff ◽  
P Deininger
Keyword(s):  
Epstein Barr Virus ◽  
Tandem Repeats ◽  
Protein Domain ◽  
Nuclear Antigen ◽  
Barr Virus ◽  
Epstein Barr ◽  
Cellular Dna ◽  
The Relationship ◽  
Simple Sequence ◽  
Tandem Arrays

We isolated clones and determined the sequence of portions of mouse and human cellular DNA which cross-hybridize strongly with the IR3 repetitive region of Epstein-Barr virus. The sequences were found to be tandem arrays of a simple sequence based on the triplet GGA, very similar to the IR3 repeat. The cellular repeats have distinct differences from the viral repeat region, however, and their sequences do not appear capable of being translated into a purely glycine-plus-alanine protein domain like the portion of the Epstein-Barr nuclear antigen coded by IR3. Although the relationship between IR3 and the cellular repeats is left unclear, the cellular repeats have many interesting features. The tandem arrays are about 1 to several kilobases long, much shorter than satellite tandem repeats and larger than other interspersed, tandem repeats. Each of the repeats is a distinct variation, perhaps diverged from a common sequence, (GGA)n. This family is present in the genomes of all species tested and appears to be a ubiquitous feature of all higher eucaryotic genomes.

scholarly journals Repeat arrays in cellular DNA related to the Epstein-Barr virus IR3 repeat.

1985 ◽  
Vol 5 (3) ◽  
pp. 457-465 ◽  
Author(s):  
M Heller ◽  
E Flemington ◽  
E Kieff ◽  
P Deininger
Keyword(s):  
Epstein Barr Virus ◽  
Tandem Repeats ◽  
Protein Domain ◽  
Nuclear Antigen ◽  
Barr Virus ◽  
Epstein Barr ◽  
Cellular Dna ◽  
The Relationship ◽  
Simple Sequence ◽  
Tandem Arrays

We isolated clones and determined the sequence of portions of mouse and human cellular DNA which cross-hybridize strongly with the IR3 repetitive region of Epstein-Barr virus. The sequences were found to be tandem arrays of a simple sequence based on the triplet GGA, very similar to the IR3 repeat. The cellular repeats have distinct differences from the viral repeat region, however, and their sequences do not appear capable of being translated into a purely glycine-plus-alanine protein domain like the portion of the Epstein-Barr nuclear antigen coded by IR3. Although the relationship between IR3 and the cellular repeats is left unclear, the cellular repeats have many interesting features. The tandem arrays are about 1 to several kilobases long, much shorter than satellite tandem repeats and larger than other interspersed, tandem repeats. Each of the repeats is a distinct variation, perhaps diverged from a common sequence, (GGA)n. This family is present in the genomes of all species tested and appears to be a ubiquitous feature of all higher eucaryotic genomes.

Role of EBNA-1 in arresting replication forks at the Epstein-Barr virus oriP family of tandem repeats

1991 ◽  
Vol 11 (12) ◽  
pp. 6268-6278
Author(s):  
V Dhar ◽  
C L Schildkraut
Keyword(s):  
Replication Fork ◽  
Epstein Barr Virus ◽  
Tandem Repeats ◽  
Simian Virus 40 ◽  
T Antigen ◽  
Nuclear Antigen ◽  
Replication Forks ◽  
Barr Virus ◽  
Epstein Barr ◽  
Sv40 Dna

The 20-member family of 30-bp tandem repeats located within the oriP region of Epstein-Barr virus (EBV) can act as a transcriptional enhancer in the presence of EBV nuclear antigen 1 (EBNA-1). A replication fork barrier and a termination site of plasmid replication in human B cells is also found within or near the EBV tandem repeats. Within each tandem repeat is a consensus binding sequence for the EBNA-1 protein that is required for extrachromosomal maintenance of oriP-containing plasmids. To investigate the factors that contribute to the arrest of replication forks and termination in the region of the family of repeats, we have used an in vitro replication system in which replication of EBV recombinant plasmids is initiated from the simian virus 40 (SV40) DNA replication origin in the presence of SV40 T antigen and soluble extracts prepared from human cells. The system can support bidirectional replication, initiating from the SV40 DNA origin with termination occurring in a region opposite the origin. Using two-dimensional agarose gel electrophoresis, we observed a barrier to replication forks in the presence of EBNA-1 in the region of the EBV repeats. Termination occurs at or near the tandem repeats in a manner similar to that observed in vivo (T.A. Gahn and C.L. Schildkraut, Cell 58:527-535, 1989). Reducing the number of repeats from 20 to 6 had little effect on the strength of the replication fork barrier. In the absence of EBNA-1, replication forks also arrested at the EBV repeats, but at a much lower efficiency. The addition of competitor DNA containing the EBV family of repeats can almost completely abolish the replication barrier produced in the presence of EBNA-1.

scholarly journals Epstein-Barr Virus (EBV) Nuclear Antigen 1 Colocalizes with Cellular Replication Foci in the Absence of EBV Plasmids

2003 ◽  
Vol 77 (6) ◽  
pp. 3824-3831 ◽  
Author(s):  
Sayuri Ito ◽  
Kazuo Yanagi
Keyword(s):  
Epstein Barr Virus ◽  
Short Pulse ◽  
Protein A ◽  
Nuclear Antigen ◽  
Metaphase Chromosomes ◽  
Barr Virus ◽  
Replication Foci ◽  
Epstein Barr ◽  
Cellular Replication ◽  
Cellular Dna

ABSTRACT Epstein-Barr virus (EBV) EBNA-1 is the only EBV-encoded protein that is essential for the once-per-cell-cycle replication and maintenance of EBV plasmids in latently infected cells. EBNA-1 binds to the oriP region of latent EBV plasmids and cellular metaphase chromosomes. In the absence of oriP-containing plasmids, EBNA-1 was highly colocalized with cellular DNA replication foci that were identified by immunostaining S-phase cells for proliferating cell nuclear antigen and replication protein A (RP-A) in combination with DNA short pulse-labeling. For the association of EBNA-1 with the cellular replication focus areas, the EBNA-1 regions of amino acids (aa) 8 to 94 and/or aa 315 to 410, but not the RP-A-interacting carboxy-terminal region, were necessary. These results suggest a new aspect of latent virus-cell interactions.

scholarly journals Differential Hyperacetylation of Histones H3 and H4 upon Promoter-Specific Recruitment of EBNA2 in Epstein-Barr Virus Chromatin

2003 ◽  
Vol 77 (14) ◽  
pp. 8166-8172 ◽  
Author(s):  
Nathalie Alazard ◽  
Henri Gruffat ◽  
Edwige Hiriart ◽  
Alain Sergeant ◽  
Evelyne Manet
Keyword(s):  
B Lymphocytes ◽  
Chromatin Immunoprecipitation ◽  
Epstein Barr Virus ◽  
Nuclear Antigen ◽  
Barr Virus ◽  
Viral Promoters ◽  
Epstein Barr ◽  
Cellular Dna ◽  
First Time

ABSTRACT Epstein-Barr virus nuclear antigen 2 (EBNA2) is a transcriptional activator involved in the immortalization of B lymphocytes by the virus. EBNA2 is targeted to the promoters of its responsive genes, via interaction with cellular DNA-binding proteins. Using chromatin immunoprecipitation assays, we show for the first time the conditional recruitment of EBNA2 on two specific viral promoters in vivo and demonstrate a correlation between this recruitment and a local change in the acetylation of histones H3 and H4, which is promoter dependent.

scholarly journals Role of EBNA-1 in arresting replication forks at the Epstein-Barr virus oriP family of tandem repeats.

1991 ◽  
Vol 11 (12) ◽  
pp. 6268-6278 ◽  
Author(s):  
V Dhar ◽  
C L Schildkraut
Keyword(s):  
Replication Fork ◽  
Epstein Barr Virus ◽  
Tandem Repeats ◽  
Simian Virus 40 ◽  
T Antigen ◽  
Nuclear Antigen ◽  
Replication Forks ◽  
Barr Virus ◽  
Epstein Barr ◽  
Sv40 Dna

The 20-member family of 30-bp tandem repeats located within the oriP region of Epstein-Barr virus (EBV) can act as a transcriptional enhancer in the presence of EBV nuclear antigen 1 (EBNA-1). A replication fork barrier and a termination site of plasmid replication in human B cells is also found within or near the EBV tandem repeats. Within each tandem repeat is a consensus binding sequence for the EBNA-1 protein that is required for extrachromosomal maintenance of oriP-containing plasmids. To investigate the factors that contribute to the arrest of replication forks and termination in the region of the family of repeats, we have used an in vitro replication system in which replication of EBV recombinant plasmids is initiated from the simian virus 40 (SV40) DNA replication origin in the presence of SV40 T antigen and soluble extracts prepared from human cells. The system can support bidirectional replication, initiating from the SV40 DNA origin with termination occurring in a region opposite the origin. Using two-dimensional agarose gel electrophoresis, we observed a barrier to replication forks in the presence of EBNA-1 in the region of the EBV repeats. Termination occurs at or near the tandem repeats in a manner similar to that observed in vivo (T.A. Gahn and C.L. Schildkraut, Cell 58:527-535, 1989). Reducing the number of repeats from 20 to 6 had little effect on the strength of the replication fork barrier. In the absence of EBNA-1, replication forks also arrested at the EBV repeats, but at a much lower efficiency. The addition of competitor DNA containing the EBV family of repeats can almost completely abolish the replication barrier produced in the presence of EBNA-1.

scholarly journals High Avidity Binding to DNA Protects Ubiquitylated Substrates from Proteasomal Degradation

2011 ◽  
Vol 286 (22) ◽  
pp. 19565-19575 ◽  
Author(s):  
Giuseppe Coppotelli ◽  
Nouman Mughal ◽  
Diego Marescotti ◽  
Maria G. Masucci
Keyword(s):  
Epstein Barr Virus ◽  
Protein Domains ◽  
Proteasomal Degradation ◽  
Nuclear Antigen ◽  
High Avidity ◽  
Dna Protection ◽  
Barr Virus ◽  
Detergent Extraction ◽  
Epstein Barr ◽  
Cellular Dna

Protein domains that act as degradation and stabilization signals regulate the rate of turnover of proteasomal substrates. Here we report that the bipartite Gly-Arg repeat of the Epstein-Barr virus (EBV) nuclear antigen (EBNA)-1 acts as a stabilization signal that inhibits proteasomal degradation in the nucleus by promoting binding to cellular DNA. Protection can be transferred by grafting the domain to unrelated proteasomal substrates and does not involve changes of ubiquitylation. Protection is also afforded by other protein domains that, similar to the Gly-Arg repeat, mediate high avidity binding to DNA, as exemplified by resistance to detergent extraction. Our findings identify high avidity binding to DNA as a portable inhibitory signal that counteracts proteasomal degradation.

scholarly journals Epstein-Barr Virus Nuclear Antigen 3C Putative Repression Domain Mediates Coactivation of the LMP1 Promoter with EBNA-2

2002 ◽  
Vol 76 (1) ◽  
pp. 232-242 ◽  
Author(s):  
Jeffrey Lin ◽  
Eric Johannsen ◽  
Erle Robertson ◽  
Elliott Kieff
Keyword(s):  
Epstein Barr Virus ◽  
Nuclear Antigen ◽  
Raji Cells ◽  
Barr Virus ◽  
Epstein Barr ◽  
Cellular Dna ◽  
High Level ◽  
Necessary And Sufficient ◽  
Two Hybrid ◽  
Repression Domain

ABSTRACT The Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA-3C) regulates virus and cell genes and is essential for EBV-mediated transformation of primary B lymphocytes. EBNA-3C associates with the cellular DNA sequence-specific transcription factors RBP-Jκ and PU.1 and coactivates the EBV LMP1 promoter with EBNA-2 in BL2 and Raji cells under conditions of restrictive growth. We now find that EBNA-3C is similar to EBNA-LP in coactivating the LMP1 promoter with EBNA-2 in non-EBV-infected Burkitt lymphoma cells under conditions of maximal cell growth, whereas the EBV Cp promoter is repressed under the same conditions. EBNA-3A and EBNA-3B coactivation are at most 40% that of EBNA-3C. The RBP-Jκ binding sites of EBNA-2 and the LMP1 promoter are not required for EBNA-3C coactivation, whereas the PU.1 site in the LMP1 promoter is required for EBNA-2-mediated activation and EBNA-3C coactivation. EBNA-3C amino acids (aa) 365 to 545, including most of the previously identified repression domain (M. Bain, R. J. Watson, P. J. Farrell, and M. J. Allday, J. Virol. 70:2481–2489, 1996), are necessary and sufficient for coactivation with wild-type EBNA-2. EBNA-3C can also coactivate with the EBNA-2 acidic activating domain; this activation does not require aa 343 to 545. These data indicate that there are at least two mechanisms by which EBNA-3C coactivates the LMP1 promoter with EBNA-2. Of the proteins that interact with EBNA-3C in a yeast two-hybrid screen, only the ubiquitin-like proteins SUMO-1 and SUMO-3/hSMT3B map to aa 365 to 545, implicating these molecules in EBNA-3C coactivation. In addition, SUMO-1 associates at a high level with EBNA-3C in lymphoblasts. Promoter coactivation by EBNA-3C is likely to be important in ensuring adequate levels of LMP1, while inhibition of the EBNA-Cp promoter under the same conditions prevents uncontrolled up-regulation of EBNA expression from a positive-feedback loop.

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