Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen 1 Mediates Episome Persistence through cis-Acting Terminal Repeat (TR) Sequence and Specifically Binds TR DNA

doi:10.1128/JVI.75.7.3250-3258.2001

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Journal of Virology, April 2001, p. 3250-3258, Vol. 75, No. 7
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.7.3250-3258.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Kaposi's Sarcoma-Associated Herpesvirus Latency-Associated Nuclear Antigen 1 Mediates Episome Persistence through cis-Acting Terminal Repeat (TR) Sequence and Specifically Binds TR DNA

Mary E. Ballestas and Kenneth M. Kaye^*

Department of Medicine, Channing Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Received 25 August 2000/Accepted 3 January 2001

Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) (also known as human herpesvirus 8) latently infects KS tumors, primaryeffusion lymphomas (PELs), and PEL cell lines. In latently infectedcells, KSHV DNA is maintained as circularized, extrachromosomalepisomes. To persist in proliferating cells, KSHV episomes mustreplicate and efficiently segregate to progeny nuclei. In uninfectedB-lymphoblastoid cells, KSHV latency-associated nuclear antigen(LANA1) is necessary and sufficient for persistence of artificialepisomes containing specific KSHV DNA. In previous work, the cis-actingsequence required for episome persistence contained KSHV terminal-repeat(TR) DNA and unique KSHV sequence. We now show that cis-actingKSHV TR DNA is necessary and sufficient for LANA1-mediated episomepersistence. Furthermore, LANA1 binds TR DNA in mobility shiftassays and a 20-nucleotide LANA1 binding sequence has been identified.Since LANA1 colocalizes with KSHV episomes along metaphase chromosomes,these results are consistent with a model in which LANA1 may bridgeTR DNA to chromosomes during mitosis to efficiently segregateKSHV episomes to progenynuclei.

^* Corresponding author. Mailing address: Department of Medicine, Channing Laboratory, Brigham and Women's Hospital, HarvardMedical School, 181 Longwood Ave., Boston, MA 02115. Phone: (617)525-4256. Fax: (617) 525-4251. E-mail: kkaye{at}rics.bwh.harvard.edu.

Journal of Virology, April 2001, p. 3250-3258, Vol. 75, No. 7
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.7.3250-3258.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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