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Journal of Virology, May 1999, p. 4385-4392, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Mapping EBNA-1 Domains Involved in Binding to Metaphase Chromosomes

Vincent Marechal,1,* Axelle Dehee,1 Roxane Chikhi-Brachet,1 Tristan Piolot,1 Maité Coppey-Moisan,2 and Jean-Claude Nicolas1

Service de Microbiologie, Hôpital Rothschild, 75571 Paris Cedex 12,1 and Laboratoire de Biochimie des Acides Nucléiques, Institut Curie, Section de recherche, 75231 Paris Cedex 05,2 France

Received 27 October 1998/Accepted 11 February 1999

The Epstein-Barr virus (EBV) genome can persist in dividing human B cells as multicopy circular episomes. Viral episomes replicate in synchrony with host cell DNA and are maintained at a relatively constant copy number for a long time. Only two viral elements, the replication origin OriP and the EBNA-1 protein, are required for the persistence of viral genomes during latency. EBNA-1 activates OriP during the S phase and may also contribute to the partition and/or retention of viral genomes during mitosis. Indeed, EBNA-1 has been shown to interact with mitotic chromatin. Moreover, viral genomes are noncovalently associated with metaphase chromosomes. This suggests that EBNA-1 may facilitate the anchorage of viral genomes on cellular chromosomes, thus ensuring proper partition and retention. In the present paper, we have investigated the chromosome-binding activity of EBV EBNA-1, herpesvirus papio (HVP) EBNA-1, and various derivatives of EBV EBNA-1, fused to a variant of the green fluorescent protein. The results show that binding to metaphase chromosomes is a common property of EBV and HVP EBNA-1. Further studies indicated that at least three independent domains (CBS-1, -2, and -3) mediate EBNA-1 binding to metaphase chromosomes. In agreement with the anchorage model, two of these domains mapped to a region that has been previously demonstrated to be required for the long-term persistence of OriP-containing plasmids.

* Corresponding author. Mailing address: Service de Microbiologie, Hôpital Rothschild, 33 Boulevard de Picpus, 75571 Paris Cedex 12, France. Phone: (33) 1 40 19 34 33. Fax: (33) 1 40 19 33 35. E-mail: vmarecha{at}infobiogen.fr.

Journal of Virology, May 1999, p. 4385-4392, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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