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

Rawlins, D R; Milman, Gregory; Hayward, S. Diane; Hayward, Gail

doi:10.1016/0092-8674(85)90282-x

Cited by 486 publications

(416 citation statements)

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“…The high transient expression by EBV-based episomal vectors may result from the multiple functions of EBNA1, such as nuclear transfer of the plasmid DNA, anchorage of the plasmid DNA to the nuclear matrix, and transcriptional enhancement. [13][14][15][16][17] The episomal replication may also contribute to the high transient transfection/expression by the EBV plasmids. However, the contribution may be small because (a) highly efficient transfection/expression can also be seen in rodent cells, in which the EBV-based plasmids cannot replicate, 32 and (b) even in human cells, the oriP-bearing plasmids replicate only once in each S phase, so unlike the SV40-based plasmid vectors, 33 the copy number of the EBV plasmids does not increase after transfection.…”

Section: Discussionmentioning

confidence: 99%

“…11,12 Through binding to oriP, EBNA1 facilitates the retention, nuclear localization, and replication of the plasmid DNA, [13][14][15][16][17] giving the EBV vector favorable features of an artificial chromosome. We have demonstrated that extremely efficient transfection can be achieved with EBV-based episomal vectors by means of electroporation in lymphoma cell lineage.…”

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confidence: 99%

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Highly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer

Harada¹,

Iwai²,

Tanaka³

et al. 2000

Cancer Gene Ther

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The present study was aimed at devising an efficient nonviral strategy for suicide gene therapy of hepatocellular carcinoma (HCC). To improve the efficiency of DNA delivery and expression, we applied Epstein-Barr virus (EBV)-based plasmid vectors instead of conventional plasmid vectors and combined them with cationic liposome (EBV/lipoplex) or polyamidoamine dendrimer (PAAD) (EBV/polyplex). When the ␤-galactosidase gene was transferred to HuH7, PLC/PRF/5, or HLE cells, Յ50-fold higher ␤-galactosidase activities were demonstrated in the cells transfected with EBV vector compared with those transfected with conventional plasmid vectors. PAAD-mediated transfection of HCC with pSES.Tk (an EBV-based vector carrying the herpes simplex virus-1 thymidine kinase gene) resulted in a marked reduction in viable cell number by the addition of ganciclovir (GCV). The HCC cells transfected with pSES.Tk/PAAD showed 100-to 1000-fold higher susceptibilities to GCV than those transfected with pS.Tk (a conventional plasmid vector carrying herpes simplex virus-1 thymidine kinase gene)/PAAD. The pSES.Tk-transfected HCC cells were effectively killed by day 9 in culture with a clinically feasible concentration of GCV (25 M), whereas the pS.Tk-transfected cells survived the culture. These results demonstrate highly efficient suicide gene transfer into various HCC cells by EBV-based plasmid vectors in vitro, suggesting the possible application of this nonviral vector system to gene therapy of HCC. Cancer Gene Therapy (2000) 7, 27-36

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Highly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer

Harada¹,

Iwai²,

Tanaka³

et al. 2000

Cancer Gene Ther

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“…DS contains four EBNA1-binding sites, albeit with lower affinity than those found in FR (Reisman, Yates et al 1985). The topology of DS is such that the four binding sites are arranged as two pairs of sites, with 21bp centerto-center spacing between each pair and 33bp center-to-center spacing between the two nonpaired internal binding sites (Figure 1.C) (Baer, Bankier et al 1984;Rawlins, Milman et al 1985). As there are 10.5bp per turn of B-form DNA, this arrangement positions two EBNA1 dimers on the same helical face of the DNA when bound to a pair of sites.…”

Section: Cis Elements Of Ebv That Contribute To Dna Replicationmentioning

confidence: 99%

“…Each monomer of EBNA1 contacts portions of DNA extending over eight base pairs and although many 16-mers known to bind EBNA1 dimers are palindromes, they need not be (Baer, Bankier et al 1984), (Rawlins, Milman et al 1985), (Wang, Lindner et al 2006). This complexity has also made it difficult to predict those sites bound by EBNA1 under physiologic conditions.…”

Section: Nih-pa Author Manuscriptmentioning

confidence: 99%

“…Initial genetic dissections of EBV identified one viral protein, Epstein-Barr Nuclear Antigen 1 (EBNA1), and one region of the viral genome, termed oriP, as being necessary and sufficient for replication of the viral plasmid (Yates, Warren et al 1984) (Lupton and Levine 1985;Reisman, Yates et al 1985), (Reisman and Sugden 1986). oriP is composed of two separable cis elements, the Family of Repeats (FR) and the Dyad Symmetry element (DS) (Baer, Bankier et al 1984;Reisman, Yates et al 1985), which both contain binding sites for EBNA1 in vitro (Rawlins, Milman et al 1985) (Harrison, Fisenne et al 1994), and in vivo (Hsieh, Camiolo et al 1993), (Niller, Glaser et al 1995) (Figure 1). FR is composed of 21 imperfect copies of a 30bp repeat and contains 20 high affinity EBNA1-binding sites (Figure 1).…”

Section: Cis Elements Of Ebv That Contribute To Dna Replicationmentioning

confidence: 99%

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The plasmid replicon of Epstein–Barr virus: Mechanistic insights into efficient, licensed, extrachromosomal replication in human cells

Lindner

Sugden

2007

Plasmid

103

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The genome of Epstein-Barr Virus (EBV) and plasmid derivatives of it are among the most efficient extrachromosomal replicons in mammalian cells. The latent origin of plasmid replication (oriP), when supplied with the viral Epstein-Barr Nuclear Antigen 1 (EBNA1) in trans, provides efficient duplication, partitioning and maintenance of plasmids bearing it. In this review, we detail what is known about the viral cis and trans elements required for plasmid replication. In addition, we describe how the cellular factors that EBV usurps are used to complement the functions of the viral constituents. Finally, we propose a model for the sequential assembly of an EBNA1-dependent origin of DNA synthesis into a pre-Replicative Complex (pre-RC), which functions by making use only of cellular enzymatic activities to carry out the replication of the viral plasmid. KeywordsoriP; EBNA1; EBV; origin; DS; Rep* General BackgroundEpstein-Barr Virus (EBV), a member of the herpesvirus family, is a surprisingly successful parasite, as are other human members of this family of viruses. It establishes a persistent, lifelong infection in greater than 90% of the world's population (Fields, Knipe et al. 2006). Primary infection by EBV causes infectious mononucleosis in some, usually adolescent people (Evans, Niederman et al. 1968). The latent cycle of the virus that follows infection is usually asymptomatic in the host, however in a small fraction of infected people, EBV contributes to several cancers including Burkitt's lymphoma, some T-cell lymphomas, Hodgkin's disease, post-transplant lymphoproliferative disease, nasopharyngeal carcinoma, and gastric carcinoma. EBV was the first human virus to be classified as a tumor virus (reviewed in Chapter 75 of (Fields, Knipe et al. 2006)) and has been studied because of its association with these diseases. EBV, in addition, has also served as a model to study DNA replication in human cells because of it's ability both to maintain its genome as an extrachromosomal replicon in infected B-cells, and to appropriate the cellular DNA replication machinery needed for its licensed replication.* To whom correspondence should be addressed: 1400 University Ave, Madison, WI 53706, Phone: 608.262.6697, Fax: 608.262.2824, Email: sugden@oncology.wisc.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Cis Elements of EBV that Contribute to DNA ReplicationThe genome of EBV is approximately 165kbp (Bloss and Sugden 1994) and resides as a nucleosome-coated nuclear plasmid in infected cells (Wensing and Farrell 2000), (Shaw, Levinger et al. 1979)...

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Circular YAC vectors containing a small mammalian origin sequence can associate with the nuclear matrix

et al. 1997

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Three different mammalian origins of DNA replication, 343, S3, and X24, have been cloned into a 15.8 kb circular yeast vector pYACneo. Subsequent transfection into HeLa cells resulted in the isolation of several stably maintained clones. Two cell lines, C343e2 and CS3e1, were found to have sequences maintained as episomes in long-term culture with a stability per generation of approximately 80%. Both episomes also contain matrix attachment region (MAR) sequences which mediate the binding of DNA to the nuclear skeleton and are thought to play a role in DNA replication. Using high salt extraction of the nucleus and fluorescent in situ hybridization, we were able to demonstrate an association of the 343 episome with the nuclear matrix, most probably through functional MAR sequences that allow an association with the nuclear matrix and associated regions containing essential replication proteins. The presence of functional MARs in small episomal sequences may facilitate the replication and maintenance of transfected DNA as an episome and improve their utility as small episomal constructs, potential microchromosomes.

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

Cited by 486 publications

References 21 publications

Highly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer

Highly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer

The plasmid replicon of Epstein–Barr virus: Mechanistic insights into efficient, licensed, extrachromosomal replication in human cells

Circular YAC vectors containing a small mammalian origin sequence can associate with the nuclear matrix

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