Fakhari Farnaz D, Dittmer Dirk P
Department of Microbiology and Immunology, The University of Oklahoma Health Science Center, Oklahoma City, Oklahoma 73104, USA.
J Virol. 2002 Jun;76(12):6213-23. doi: 10.1128/jvi.76.12.6213-6223.2002.
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.
分为潜伏或裂解生命周期是所有疱疹病毒科的基本特征。就卡波西肉瘤相关疱疹病毒（KSHV，即人类疱疹病毒8型）而言，潜伏基因与细胞自主转化有关，而某些裂解基因则通过旁分泌机制营造有利于肿瘤生长的环境。为了探究KSHV转录情况，我们设计并验证了一种高通量、高特异性、高灵敏度的实时定量逆转录PCR阵列。这种新方法适用于多种人类病原体。其首次应用表明，病毒重新激活后，KSHV的LANA、v-细胞周期蛋白和v-FLIP的mRNA水平在任何时候都不会增加。LANA-2/vIRF-3的mRNA同样对病毒重新激活有抗性。相比之下，其他所有潜伏或裂解信息都被诱导。因此，LANA、v-FLIP、v-细胞周期蛋白和LANA-2构成了KSHV基因组中一组独特调控的转录本。
