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Journal of Virology, June 2002, p. 6213-6223, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6213-6223.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Charting Latency Transcripts in Kaposi's Sarcoma-Associated Herpesvirus by Whole-Genome Real-Time Quantitative PCR

Farnaz D. Fakhari and Dirk P. Dittmer*

Department of Microbiology and Immunology, The University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104

Received 6 December 2001/ Accepted 12 March 2002

The division into a latent or lytic life cycle is fundamental to all herpesviridae. In the case of Kaposi's sarcoma-associated herpesvirus (KSHV) (human herpesvirus 8), latent genes have been implicated in cell autonomous transformation, while certain lytic genes procure a tumor friendly milieu through paracrine mechanism. To query KSHV transcription, we devised and validated a high-throughput, high-specificity, high-sensitivity, real-time quantitative reverse transcription-PCR array. This novel methodology is applicable to many human pathogens. Its first use demonstrated that the mRNA levels for KSHV LANA, v-cyclin, and v-FLIP do not increase at any time after viral reactivation. The mRNA for LANA-2/vIRF-3 is similarly resistant to viral reactivation. In contrast, every other latent or lytic message was induced. Hence, LANA, v-FLIP, v-cyclin, and LANA-2 constitute a group of uniquely regulated transcripts in the KSHV genome.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, The University of Oklahoma Health Science Center, 940 Stanton L. Young Blvd., Oklahoma City, OK 73104. Phone: (405) 271-2690. Fax: (405) 271-3117. E-mail: dirk-dittmer{at}ouhsc.edu.

Journal of Virology, June 2002, p. 6213-6223, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6213-6223.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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