Replication of latent Epstein‐Barr virus genomes in normal and malignant lymphoid cells

Adams, Alice; Pozos, Tamara C.; Purvey, Helen V.

doi:10.1002/ijc.2910440331

Cited by 65 publications

(70 citation statements)

References 29 publications

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“…A second origin of DNA synthesis has been identified in the EBV genome, which also functions extrachromosomally and is not discrete but does support licensed DNA synthesis (Fig. 1A) (12)(13)(14)(15). These latter properties make this origin, which we term ''Raji ori,'' similar to well studied mammalian chromosomal origins such as the DHFR (16) and ␤-globin (17,18).…”

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confidence: 90%

Identifying a property of origins of DNA synthesis required to support plasmids stably in human cells

Wang

Sugden

2008

Proc. Natl. Acad. Sci. U.S.A.

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The plasmid origin of replication, oriP, of Epstein-Barr Virus (EBV) was identified in an assay to detect autonomously replicating sequences (ARSs) in human cells. Raji ori, a second origin in EBV, functions in vivo but fails in long-term ARS assays. We examined the initiating element, DS, within oriP and Raji ori to resolve this paradox. DS, but not Raji ori, binds EBNA1; whereas both act as ARSs in short-term assays, with DS being more efficient, only DS can act as an ARS in long-term assays. Surprisingly, we found that DS supported the establishment of a plasmid with Raji ori in cis and that after deletion of DS, Raji ori could now act as an ARS in the long term. This finding explains the frequent failure of ARS assays in mammalian cells. More origins can initially act as ARSs than can be established. We identified one requirement for ARSs to be established: They must function efficiently enough initially to generate a wide distribution of numbers of plasmids per cell. Only the cells that have more than a threshold number of plasmids can survive selections imposed on the cells to retain these replicons.autonomously replicating sequence ͉ DNA replication ͉ Epstein-Barr virus T he study of DNA replication in the budding yeast Saccharomyces cerevisiae has provided much of the foundation of our understanding of DNA replication in eukaryotic cells. In S. cerevisiae, assays for autonomously replicating sequences (ARS) were used to identify the origins of DNA synthesis after their introduction into plasmids having a functional centromere (CEN) (1-5). Unlike ARSs in yeast, origins of DNA synthesis in mammalian cells are poorly understood. Parallel experiments in mammalian cells have not been the standard initially used to identify chromosomal origins of DNA synthesis. One difficulty in developing a mammalian ARS assay, for example, is imposed by the size of mammalian centromeres making their use difficult.One mammalian origin of DNA synthesis that was identified in an ARS assay in human cells is the Dyad symmetry (DS) of Epstein-Barr virus (EBV) (6, 7). DS is a discrete, licensed origin that functions in a variety of mammalian hosts in the presence of the viral trans-acting element EBNA1 (Epstein-Barr nuclear antigen 1) (8). It requires the family of repeats (FR) from EBV in cis and EBNA1 in trans to provide its segregation mechanism to the replicon (9-11). FR and EBNA1 thus functionally replace the requirement in yeast for a CEN element in ARS assays. A second origin of DNA synthesis has been identified in the EBV genome, which also functions extrachromosomally and is not discrete but does support licensed DNA synthesis (Fig. 1A) (12-15). These latter properties make this origin, which we term ''Raji ori,'' similar to well studied mammalian chromosomal origins such as the DHFR (16) and ␤-globin (17, 18). Raji ori was identified by two-dimensional gel electrophoresis and in an assay termed ''single molecule analysis of replicated DNA'' (SMARD) and was found both to be the predominant origin of DNA synthesis used in t...

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confidence: 90%

Identifying a property of origins of DNA synthesis required to support plasmids stably in human cells

Wang

Sugden

2008

Proc. Natl. Acad. Sci. U.S.A.

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“…[1][2][3] Once the linear viral DNA enters the cell it recircularizes and subsequently establishes a latent pattern of replication in the nucleus of the infected cell for the life of the individual. The viral genome is then replicated only once per cell division cycle [4][5][6] by the host's DNA replication machinery in synchrony with the host's genome. The large viral episome has been found to maintain a stable copy number of between five and 50 per cell nucleus in Burkitt's lymphoma (BL) and lymphoblastoid cell lines (LCL).…”

Section: Correspondence: J-mh Vosmentioning

confidence: 99%

An enhanced EBNA1 variant with reduced IR3 domain for long-term episomal maintenance and transgene expression of oriP-based plasmids in human cells

Wendelburg

Vos

1998

Gene Ther

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The latent replication of oriP-based plasmids in human IR3-deleted, form, as well as a new EBNA1 isoform cloned cells depends on the viral oriP-binding transactivator from Raji. The results of a 6-month study indicate that the EBNA1. In this report, the effect of the internal repeat 3 isoforms of EBNA1 differ with respect to their efficiency of (IR3 or GlyAla repeat) domain of EBNA1 on long-term plasmid maintenance. While the EBNA-1 Raji encoding maintenance and transgene expression of OriP-based plasmid was the most stable, the oriP-based vector plasmids was examined in dividing human cells. To assess expressing the truncated EBNA1 (IR3del) gene was lost at the potential contribution of different isoforms of EBNA1 a much higher rate than those transducing full size specifically, the long-term stability of oriP-based plasmids EBNA1s. In parallel, long-term reporter gene expression in was determined after stable transfection of various CMVvarious human B cell lines was shown to persist at the driven EBNA1 genes in EBV-negative human B cells. Epihighest level with the oriP-based Raji EBNA-1 construct. some copy number was quantified using a novel sensitive These results show that the GlyAla domain can positively assay based on human mitochondrial DNA as an internal influence long-term plasmid stability and episomal transextrachromosomal control. Using this assay, the standard gene expression. B95.8-derived EBNA1 was compared with its truncated,

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“…16 Upon reactivation, EBV can productively infect oropharyngeal epithelium, leading to infectious virus production and transmission. 19 In latent infection, EBV genomic DNA exists as an episome, replicating only once during S phase 20 and partitioning accurately into daughter cells during the mitotic phase. 21 In lytic state, the EBV genomic DNA is linear.…”

Section: Introductionmentioning

confidence: 99%

Regulation of cellular and viral protein expression by the Epstein-Barr virus transcriptional regulator Zta: Implications for therapy of EBV associated tumors

Chen¹,

Li²,

Guo³

2009

Cancer Biology & Therapy

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Replication of latent Epstein‐Barr virus genomes in normal and malignant lymphoid cells

Cited by 65 publications

References 29 publications

Identifying a property of origins of DNA synthesis required to support plasmids stably in human cells

Identifying a property of origins of DNA synthesis required to support plasmids stably in human cells

An enhanced EBNA1 variant with reduced IR3 domain for long-term episomal maintenance and transgene expression of oriP-based plasmids in human cells

Regulation of cellular and viral protein expression by the Epstein-Barr virus transcriptional regulator Zta: Implications for therapy of EBV associated tumors

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