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Journal of Virology, July 2000, p. 6207-6212, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Kaposi's Sarcoma-Associated Herpesvirus Open Reading Frame 50/Rta Protein Activates the Entire Viral Lytic Cycle in the HH-B2 Primary Effusion Lymphoma Cell Linedagger

Lyndle Gradoville,1 Jennifer Gerlach,2 Elizabeth Grogan,1 Duane Shedd,1 Sarah Nikiforow,3 Craig Metroka,4 and George Miller1,5,6,*

Departments of Pediatrics,1 Molecular Biophysics and Biochemistry,5 Epidemiology and Public Health,6 Cell Biology,2 and Immunobiology,3 Yale University School of Medicine, New Haven, Connecticut 06520, and St. Luke's-Roosevelt Hospital Center, New York, New York4

Received 18 January 2000/Accepted 11 April 2000

Rta, the gene product of Kaposi's sarcoma-associated herpesvirus (KSHV) encoded mainly in open reading frame 50 (ORF50), is capable of activating expression of viral lytic cycle genes. What was not demonstrated in previous studies was whether KSHV Rta was competent to initiate the entire viral lytic life cycle including lytic viral DNA replication, late-gene expression with appropriate kinetics, and virus release. In HH-B2, a newly established primary effusion lymphoma (PEL) cell line, KSHV ORF50 behaved as an immediate-early gene and autostimulated its own expression. Expression of late genes, ORF65, and K8.1 induced by KSHV Rta was eliminated by phosphonoacetic acid, an inhibitor of viral DNA polymerase. Transfection of KSHV Rta increased the production of encapsidated DNase-resistant viral DNA from HH-B2 cells. Thus, introduction of an ORF50 expression plasmid is sufficient to drive the lytic cycle to completion in cultured PEL cells.

* Corresponding author. Mailing address: Department of Pediatrics, Yale University School of Medicine, 333 Cedar St., New Haven, CT 06520. Phone: (203) 785-4758. Fax: (203) 785-6961. E-mail: George.Miller{at}yale.edu.

dagger This paper is dedicated to the memory of Elizabeth Grogan, our close colleague and collaborator for many years, who isolated the HH-B2 cell line.

Journal of Virology, July 2000, p. 6207-6212, Vol. 74, No. 13
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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