Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein-Barr virus

Hammerschmidt, Wolfgang; Sugden, Bill

doi:10.1016/0092-8674(88)90028-1

Cited by 362 publications

(390 citation statements)

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“…Thick lines indicate important coding or regulatory sequences within the regions probed. The following probes were used: (i) an EcoRI-BamHI subfragment from the BamHI C fragment (nucleotides 7315 to 13 215) covering oriP, the latent origion of EBV replication (Yates et al, 1984); (ii) the BamHI W fragment containing a B cell-specific'enhancer (W ENH) (Ricksten et al, 1988); (iii) the BamHI H fragment partly covering EBNA 2 coding sequences and containing oriLyt, the lytic origin of EBV replication (Hammerschmidt & Sugden, 1988); (iv) the BamHI M fragment containing coding sequences for early antigens (open reading frames BMRF 1 and BMRF 2); (v) the BamHI E fragment encoding EBNA 3, 4 and 6; (vi) the BamHI K fragment.encoding EBNA l ; (vii) a 2.4 kb MluI subfragment of the EcoDHET fragment covering the coding sequences of LMP (nueleotides 167129 to 169 566); (viii) a 0.8 kb BglI fragment (nucleotides 169 449 to 170290); or (ix) a XhoI-EcoRI fragment (nucleotides 169 424 to 172 284) for the analysis of regulatory sequences (LRS) of the LMP gene (F~thraeus et al, 1990). (ii) Immunoblotting.…”

Section: Methodsmentioning

confidence: 99%

Host cell phenotype-dependent methylation patterns of Epstein--Barr virus DNA

Minárovits

Minarovits-Kormuta

Ehlin‐Henriksson

et al. 1991

Journal of General Virology

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We have shown previously that the EBNA 1 and latent membrane protein encoding regions of the EpsteinBarr virus (EBV) genome are highly methylated at CCGG sequences in the Burkitt's lymphoma (BL)-derived cell line Rael, but are unmethylated in a lymphoblastoid cell line (LCL) harbouring the same virus. To examine whether this is a regular phenomenon, we compared the methylation patterns of selected regions (BamHl C, W, H, M, E, K and N fragments) of EBV DNA in representative EBVcarrying cell types of normal and neoplastic origin. Analysis of HpalI and MspI cleavage patterns showed that all probed regions were highly methylated in all six BL biopsy samples, but hypomethylated in the four LCLs immortalized by the virus. EBV DNA was also highly methylated in the nude mouse-passaged C15 nasopharyngeal carcinoma strain and partially methylated in the C18 strain. Eight BL lines propagated in vitro, ranging from a typical BL group I to a more LCLlike group III phenotype, showed heterogeneous levels of methylation. Rael, the only stable group I cell line, carried highly methylated viral genomes. The other cell lines, which have drifted to an LCL-like blastic phenotype to various degrees, showed more moderate or low viral DNA methylation. Two sublines of the BL cell line Jijoye, which could be classified as groups II and III, respectively, showed a corresponding difference in EBV DNA methylation. To assess the possible influence of hypomethylated linear EBV DNA molecules produced in lytically infected cells, the virus producer P3HR-1 and Jijoye M13 lines were compared for DNA methylation before and after treatment with phosphonoformic acid (PFA), an inhibitor of the viral DNA polymerase. PFA treatment resulted in a shift towards a more methylated pattern in both regions (BamHI W and E) assayed, but had no effect on virus non-producer lines (Rael, CB-M1-RaI-STO and Jijoye p79).

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Section: Methodsmentioning

confidence: 99%

Host cell phenotype-dependent methylation patterns of Epstein--Barr virus DNA

Minárovits

Minarovits-Kormuta

Ehlin‐Henriksson

et al. 1991

Journal of General Virology

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“…The PCR products were digested with DpnI to remove residual template DNA. Approximately 200 ng of the linear PCR product was electroporated into recombinase-induced E. coli DH10B electrocompetent cells (harboring pREDE5 and EBV BAC), which were prepared as described previously (3). Electrotransformation was performed using a Bio-Rad Gene Pulser II electroporation system (0.1-cm cuvette, 1.6 kV, 25 microfarads, 200 ohms, 50 l of cells).…”

Section: Methodsmentioning

confidence: 99%

“…In latently infected cells, only limited numbers of viral genes are expressed (2) with no production of virus particles. Lytic infection differs in that multiple rounds of replication are initiated within the oriLyt region of the EBV genome (3). One of the first detectable changes is expression of the BZLF1 gene product, which is also called Zta or ZEBRA.…”

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confidence: 99%

Epstein-Barr Virus Polymerase Processivity Factor Enhances BALF2 Promoter Transcription as a Coactivator for the BZLF1 Immediate-Early Protein

Nakayama

Murata

Murayama

et al. 2009

Journal of Biological Chemistry

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The Epstein-Barr virus (EBV) BMRF1 protein is an essential replication protein acting at viral replication forks as a viral DNA polymerase processivity factor, whereas the BALF2 protein is a singlestranded DNA-binding protein that also acts at replication forks and is most abundantly expressed during viral productive replication. Here we document that the BMRF1 protein evidently enhances viral BZLF1 transcription factor-mediated transactivation of the BALF2 gene promoter. Mutagenesis and electrophoretic mobility shift assays demonstrated the BALF2 promoter to harbor two BZLF1 protein-binding sites (BZLF1-responsive elements). Direct binding of the BZLF1 protein to BZLF1-responsive elements and physical interaction between BZLF1 and BMRF1 proteins are prerequisite for the BMRF1 protein up-regulation of the BALF2 gene promoter. A monomeric mutant, C95E, which is defective in homodimerization, could still interact and enhance BZLF1-mediated transactivation. Furthermore although EBV protein kinase phosphorylates BMRF1 protein extensively, it turned out that phosphorylation of the protein by the kinase is inhibitory to the enhancement of the BZLF1-mediated transactivation of BALF2 promoter. Exogenous expression of BMRF1 protein augmented BALF2 expression in HEK293 cells harboring the EBV genome but lacking BMRF1 and BALF5 genes, demonstrating functions as a transcriptional regulator in the context of viral infection. Overall the BMRF1 protein is a multifunctional protein that cannot only act as a DNA polymerase processivity factor but also enhances BALF2 promoter transcription as a coactivator for the BZLF1 protein, regulating the expression level of viral singlestranded DNA-binding protein.

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“…26. To obtain virus stocks, the cell lines were transiently transfected with expression plasmids encoding BZLF1 (27) and BALF4 (6) to induce EBV's lytic cycle. Four days p.i., supernatants were harvested and filtered through 1.2-m pore filters.…”

Section: Methodsmentioning

confidence: 99%

Transcriptional activation by EBV nuclear antigen 1 is essential for the expression of EBV's transforming genes

Altmann

Pich

Ruiss

et al. 2006

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

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126

123

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EBV is a paradigm for human tumor viruses because, although it infects most people benignly, it also can cause a variety of cancers. Both in vivo and in vitro, EBV infects B lymphocytes in G0, induces them to become blasts, and can maintain their proliferation in cell culture or in vivo as tumors. How EBV succeeds in these contrasting cellular environments in expressing its genes that control the host has not been explained. We have genetically dissected the EBV nuclear antigen 1 (EBNA1) gene that is required for replication of the viral genome, to elucidate its possible role in the transcription of viral genes. Strikingly, EBNA1 is essential to drive transcription of EBV's transforming genes after infection of primary B lymphocytes.transcriptional regulation ͉ high-mobility group A protein ͉ HMGA1a

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Identification and characterization of oriLyt, a lytic origin of DNA replication of Epstein-Barr virus

Cited by 362 publications

References 23 publications

Host cell phenotype-dependent methylation patterns of Epstein--Barr virus DNA

Host cell phenotype-dependent methylation patterns of Epstein--Barr virus DNA

Epstein-Barr Virus Polymerase Processivity Factor Enhances BALF2 Promoter Transcription as a Coactivator for the BZLF1 Immediate-Early Protein

Transcriptional activation by EBV nuclear antigen 1 is essential for the expression of EBV's transforming genes

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