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Journal of Virology, December 2005, p. 15099-15106, Vol. 79, No. 24
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.24.15099-15106.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Transcriptional Analysis of Latent and Inducible Kaposi's Sarcoma-Associated Herpesvirus Transcripts in the K4 to K7 Region{dagger}

Jennifer L. Taylor, Heather N. Bennett, Beth A. Snyder, Patrick S. Moore, and Yuan Chang*

Molecular Virology Program, University of Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, Pennsylvania 15213

Received 22 April 2005/ Accepted 23 June 2005

Kaposi's sarcoma-associated herpesvirus (KSHV) is a gamma-2 herpesvirus with a genome containing a long unique coding region (LUR) flanked by GC-rich terminal repeat sequences. The LUR encodes approximately 90 annotated open reading frames (ORFs) with complex patterns of gene expression during viral latency, reactivation, and de novo infection. To identify unannotated KSHV genes, we examined the region between 21,500 and 30,000 bp of the KSHV LUR, representing approximately 8.5 kb of sequence. This region encodes seven known single-exon ORFs (K4, K4.1, K4.2, K5, K6, K7, and PAN), but previous computer analyses have failed to identify additional likely genes in the remaining 5.2 kb. We identified four novel transcripts using Northern blotting, phage library screening, and 5' rapid amplification of cDNA ends analysis in the region between ORFs K4.2 and K7. In vitro analysis of KSHV-infected primary effusion lymphoma cell lines in the presence of 12-O-tetradecanoylphorbol-13-acetate and phosphonoformic acid suggests that one latent transcript is coterminal with the previously annotated K3 gene encoding an ubiquitin-ligase known to downregulate major histocompatibility complex class I expression. This alternatively spliced transcript may contribute to KSHV adaptive immune evasion during latent infection. Other transcripts are inducible, including a 6.1-kb transcript that is the largest transcript found in the KSHV genome to date.

* Corresponding author. Mailing address: Hillman Cancer Center, Molecular Virology Program, University of Pittsburgh Cancer Institute, 5117 Centre Avenue, Suite 1.8, Pittsburgh, PA 15213. Phone: (412) 623-7721. Fax: (412) 623-7715. E-mail: yc70{at}pitt.edu.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, December 2005, p. 15099-15106, Vol. 79, No. 24
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.24.15099-15106.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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