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Efficient replication of Epstein-Barr virus-derived plasmids requires tethering by EBNA1 to host chromosomes.
J Virol. 2013 Dec; 87(23):13020-8.JV

Abstract

The EBNA1 protein of Epstein-Barr virus enables plasmids carrying oriP both to duplicate and to segregate efficiently in proliferating cells. EBNA1 recruits the origin recognition complex (ORC) to establish a replication origin at one element of oriP, DS (dyad symmetry); at another element, FR (family of repeats), EBNA1 binds to an array of sites from which it tethers plasmids to host chromosomes for mitotic stability. We report experiments leading to the conclusion that tethering by EBNA1 to host chromosomes is also needed within interphase nuclei in order for plasmids to be replicated efficiently from oriP. The DNA-binding domain of EBNA1, which lacks chromosome-binding ability, was found to support weak, DS-specific replication in HEK293 cells after transient transfection, being 17% as active as wild-type EBNA1. The low efficiency of replication was not due to the failure of the DNA-binding domain to retain plasmids within nuclei, because plasmids were recovered in similar amounts and entirely from the nuclear fraction of these transiently transfected cells. A derivative of EBNA1 with its chromosome-tethering domains replaced by a 22-amino-acid nucleosome-binding domain was fully active in supporting oriP functions. The implication is that EBNA1's DNA-binding domain is able to recruit ORC to DS, but either this step or subsequent replication is only efficient if the plasmid is tethered to a host chromosome. Finally, with some cell lines, DS can hardly support even transient plasmid replication without FR. A loss of plasmids lacking FR from nuclei cannot account for this requirement, suggesting that the stronger tethering to chromosomes by FR is needed for plasmid replication within the nuclei of such cells.

Authors+Show Affiliations

Roswell Park Cancer Institute, Department of Cancer Genetics, Buffalo, New York, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

24067969

Citation

Hodin, Theresa L., et al. "Efficient Replication of Epstein-Barr Virus-derived Plasmids Requires Tethering By EBNA1 to Host Chromosomes." Journal of Virology, vol. 87, no. 23, 2013, pp. 13020-8.
Hodin TL, Najrana T, Yates JL. Efficient replication of Epstein-Barr virus-derived plasmids requires tethering by EBNA1 to host chromosomes. J Virol. 2013;87(23):13020-8.
Hodin, T. L., Najrana, T., & Yates, J. L. (2013). Efficient replication of Epstein-Barr virus-derived plasmids requires tethering by EBNA1 to host chromosomes. Journal of Virology, 87(23), 13020-8. https://doi.org/10.1128/JVI.01606-13
Hodin TL, Najrana T, Yates JL. Efficient Replication of Epstein-Barr Virus-derived Plasmids Requires Tethering By EBNA1 to Host Chromosomes. J Virol. 2013;87(23):13020-8. PubMed PMID: 24067969.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Efficient replication of Epstein-Barr virus-derived plasmids requires tethering by EBNA1 to host chromosomes. AU - Hodin,Theresa L, AU - Najrana,Tanbir, AU - Yates,John L, Y1 - 2013/09/25/ PY - 2013/9/27/entrez PY - 2013/9/27/pubmed PY - 2014/1/9/medline SP - 13020 EP - 8 JF - Journal of virology JO - J Virol VL - 87 IS - 23 N2 - The EBNA1 protein of Epstein-Barr virus enables plasmids carrying oriP both to duplicate and to segregate efficiently in proliferating cells. EBNA1 recruits the origin recognition complex (ORC) to establish a replication origin at one element of oriP, DS (dyad symmetry); at another element, FR (family of repeats), EBNA1 binds to an array of sites from which it tethers plasmids to host chromosomes for mitotic stability. We report experiments leading to the conclusion that tethering by EBNA1 to host chromosomes is also needed within interphase nuclei in order for plasmids to be replicated efficiently from oriP. The DNA-binding domain of EBNA1, which lacks chromosome-binding ability, was found to support weak, DS-specific replication in HEK293 cells after transient transfection, being 17% as active as wild-type EBNA1. The low efficiency of replication was not due to the failure of the DNA-binding domain to retain plasmids within nuclei, because plasmids were recovered in similar amounts and entirely from the nuclear fraction of these transiently transfected cells. A derivative of EBNA1 with its chromosome-tethering domains replaced by a 22-amino-acid nucleosome-binding domain was fully active in supporting oriP functions. The implication is that EBNA1's DNA-binding domain is able to recruit ORC to DS, but either this step or subsequent replication is only efficient if the plasmid is tethered to a host chromosome. Finally, with some cell lines, DS can hardly support even transient plasmid replication without FR. A loss of plasmids lacking FR from nuclei cannot account for this requirement, suggesting that the stronger tethering to chromosomes by FR is needed for plasmid replication within the nuclei of such cells. SN - 1098-5514 UR - https://www.unboundmedicine.com/medline/citation/24067969/Efficient_replication_of_Epstein_Barr_virus_derived_plasmids_requires_tethering_by_EBNA1_to_host_chromosomes_ L2 - http://jvi.asm.org/cgi/pmidlookup?view=long&pmid=24067969 DB - PRIME DP - Unbound Medicine ER -