This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Norio, P.
Right arrow Articles by Yates, J. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Norio, P.
Right arrow Articles by Yates, J. L.

 Previous Article  |  Next Article 

Journal of Virology, September 2000, p. 8563-8574, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Initiation of DNA Replication within oriP Is Dispensable for Stable Replication of the Latent Epstein-Barr Virus Chromosome after Infection of Established Cell Lines

Paolo Norio,1 Carl L. Schildkraut,1,* and John L. Yates2,*

Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461,1 and Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 142632

Received 14 February 2000/Accepted 20 June 2000

The 165-kb circularized chromosome of Epstein-Barr virus (EBV) is replicated in latently infected cells once per cell cycle by host proteins during S phase. Replication initiates at multiple sites on latent EBV chromosomes, including within a 1.8-kb region called oriP, which can provide both replication and stabilization for recombinant plasmids in the presence of the EBV-encoded protein, EBNA-1. Replication initiates at or near the dyad symmetry component (DS) of oriP, which depends on multiple EBNA-1 binding sites for activity. To test the importance of the replication function of oriP, the DS was deleted from the viral genome. EBV mutants lacking the DS and carrying a selectable gene could establish latent infections in BL30 cells, in which circular, mutant viral chromosomes were stably maintained. Analysis of replication fork movement using two-dimensional gel electrophoresis showed that the deletion of the DS reduced the initiation events to an undetectable level within the oriP region so that this segment was replicated exclusively by forks entering the region from either direction. A significant slowing or stalling of replication forks that occurs normally at the approximate position of the DS was also eliminated by deletion of the DS. The results confirm the DS as both a replication origin and a place where replication forks pause. Since the replication function of oriP is dispensable at least in certain cell lines, the essential role of EBNA-1 for infection of these cell lines is likely to be that of stabilizing the EBV chromosome by associating with the 30-bp repeats of oriP. The results also imply that in established cell lines, the EBV chromosome can be efficiently replicated entirely from origins that are activated by cellular factors. Presumably, initiation of replication at the DS, mediated by EBNA-1, is important for the natural life cycle of EBV, perhaps in establishing latent infections of normal B cells.

* Corresponding author. Mailing address for Carl L. Schildkraut: Dept. of Cell Biology (CH416), Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-2097. Fax: (718) 430-8574. E-mail: schildkr{at}aecom.yu.edu. Mailing address for John L. Yates: Department of Genetics, Roswell Park Cancer Institute, Elm and Carlton St., Buffalo, NY 14263. Phone: (716) 845-8964. Fax: (716) 845-8449. E-mail: Yates{at}sc3101.med.buffalo.edu.

Journal of Virology, September 2000, p. 8563-8574, Vol. 74, No. 18
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Wang, C.-Y., Sugden, B. (2008). Identifying a property of origins of DNA synthesis required to support plasmids stably in human cells. Proc. Natl. Acad. Sci. USA 105: 9639-9644 [Abstract] [Full Text]  
  • Verma, S. C., Choudhuri, T., Robertson, E. S. (2007). The Minimal Replicator Element of the Kaposi's Sarcoma-Associated Herpesvirus Terminal Repeat Supports Replication in a Semiconservative and Cell-Cycle-Dependent Manner. J. Virol. 81: 3402-3413 [Abstract] [Full Text]  
  • Deng, Z., Atanasiu, C., Burg, J. S., Broccoli, D., Lieberman, P. M. (2003). Telomere Repeat Binding Factors TRF1, TRF2, and hRAP1 Modulate Replication of Epstein-Barr Virus OriP. J. Virol. 77: 11992-12001 [Abstract] [Full Text]  
  • Ritzi, M., Tillack, K., Gerhardt, J., Ott, E., Humme, S., Kremmer, E., Hammerschmidt, W., Schepers, A. (2003). Complex protein-DNA dynamics at the latent origin of DNA replication of Epstein-Barr virus. J. Cell Sci. 116: 3971-3984 [Abstract] [Full Text]  
  • Collins, C. M., Medveczky, M. M., Lund, T., Medveczky, P. G. (2002). The terminal repeats and latency-associated nuclear antigen of herpesvirus saimiri are essential for episomal persistence of the viral genome. J. Gen. Virol. 83: 2269-2278 [Abstract] [Full Text]  
  • Rose, C., Green, M., Webber, S., Kingsley, L., Day, R., Watkins, S., Reyes, J., Rowe, D. (2002). Detection of Epstein-Barr Virus Genomes in Peripheral Blood B Cells from Solid-Organ Transplant Recipients by Fluorescence In Situ Hybridization. J. Clin. Microbiol. 40: 2533-2544 [Abstract] [Full Text]  
  • Koons, M. D., Van Scoy, S., Hearing, J. (2001). The Replicator of the Epstein-Barr Virus Latent Cycle Origin of DNA Replication, oriP, Is Composed of Multiple Functional Elements. J. Virol. 75: 10582-10592 [Abstract] [Full Text]  
  • White, R. E., Wade-Martins, R., James, M. R. (2001). Sequences Adjacent to oriP Improve the Persistence of Epstein-Barr Virus-Based Episomes in B Cells. J. Virol. 75: 11249-11252 [Abstract] [Full Text]  
  • Shirakata, M., Imadome, K.-I., Okazaki, K., Hirai, K. (2001). Activation of TRAF5 and TRAF6 Signal Cascades Negatively Regulates the Latent Replication Origin of Epstein-Barr Virus through p38 Mitogen-Activated Protein Kinase. J. Virol. 75: 5059-5068 [Abstract] [Full Text]  
  • Kanda, T., Otter, M., Wahl, G. M. (2001). Coupling of Mitotic Chromosome Tethering and Replication Competence in Epstein-Barr Virus-Based Plasmids. Mol. Cell. Biol. 21: 3576-3588 [Abstract] [Full Text]  
  • Haan, K. M., Aiyar, A., Longnecker, R. (2001). Establishment of Latent Epstein-Barr Virus Infection and Stable Episomal Maintenance in Murine B-Cell Lines. J. Virol. 75: 3016-3020 [Abstract] [Full Text]  
  • Chaudhuri, B., Xu, H., Todorov, I., Dutta, A., Yates, J. L. (2001). Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus. Proc. Natl. Acad. Sci. USA 98: 10085-10089 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»