Charting latency transcripts in Kaposi's sarcoma-associated herpesvirus by whole-genome real-time quantitative PCR

FD Fakhari, DP Dittmer - Journal of virology, 2002 - Am Soc Microbiol
FD Fakhari, DP Dittmer
Journal of virology, 2002Am Soc Microbiol
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 …
Abstract
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.
American Society for Microbiology