Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator
Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator
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DOI：
10.1128/jvi.77.7.4205-4220.2003
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发表时间：
2003-04-01
影响因子：
5.4
通讯作者：
Jung, JU
中科院分区：
文献类型：
--
作者：
Nakamura, H;Lu, M;Jung, JU
疱疹病毒生命周期的一个重要步骤是从潜伏期到裂解性再激活的转换。为了研究卡波西肉瘤相关疱疹病毒（KSHV）的生命周期，我们在KSHV感染的原发积液淋巴瘤细胞中建立了基因表达系统。该系统采用flp介导的高效重组和四环素诱导表达。Rta转录激活因子作为KSHV裂解再激活的分子开关，有效地整合在Flp重组靶点下游，其表达受到四环素的严格控制。与TPA刺激一样，Rta的异位表达有效地诱导了病毒复制的完整周期，包括有序的KSHV基因表达程序和传染性病毒子代的产生。Rta介导的裂解基因表达的一个显著特征是，Rta诱导KSHV基因表达的能力比TPA刺激更强、更有效，这表明Rta在KSHV裂解基因表达中起着核心、主导作用。因此，我们的流线型基因表达系统不仅为研究病毒基因产物对KSHV整体基因表达和复制的影响提供了一种新的手段，而且还为了解病毒的自然再激活过程提供了一种新的方法。
An important step in the herpesvirus life cycle is the switch from latency to lytic reactivation. In order to study the life cycle of Kaposi's sarcoma-associated herpesvirus (KSHV), we developed a gene expression system in KSHV-infected primary effusion lymphoma cells. This system uses Flp-mediated efficient recombination and tetracycline-inducible expression. The Rta transcriptional activator, which acts as a molecular switch for lytic reactivation of KSHV, was efficiently integrated downstream of the Flp recombination target site, and its expression was tightly controlled by tetracycline. Like stimulation with tetradecanoyl phorbol acetate (TPA), the ectopic expression of Rta efficiently induced a complete cycle of viral replication, including a well-ordered program of KSHV gene expression and production of infectious viral progeny. A striking feature of Rta-mediated lytic gene expression was that Rta induced KSHV gene expression in a more powerful and efficient manner than TPA stimulation, indicating that Rta plays a central, leading role in KSHV lytic gene expression. Thus, our streamlined gene expression system provides a novel means not only to study the effects of viral gene products on overall KSHV gene expression and replication, but also to understand the natural viral reactivation process.