Gauliard Nicolas, Billecocq Agnès, Flick Ramon, Bouloy Michèle
Institut Pasteur, Unité de Génétique Moléculaire des Bunyaviridés, 25 rue du Dr Roux 75724 Paris cedex 15, France.
Virology. 2006 Jul 20;351(1):170-9. doi: 10.1016/j.virol.2006.03.018. Epub 2006 Apr 21.
Rift Valley fever virus (RVFV) (Phlebovirus, Bunyaviridae) possesses a genome composed of three negative-stranded RNA molecules. Each segment contains 3' and 5' noncoding regions with terminal complementary sequences forming a panhandle structure. We showed that transcription-replication of the L, M and S segments is regulated, and we established a minigenome rescue system expressing a CAT reporter to investigate the role of the noncoding regions in this process. The L, M and S segment-based minigenomes were shown to drive bona fide transcription and replication and to express variable levels of CAT reporter, indicating differential promoter activities within the noncoding sequences. In addition, we found a good correlation between the relative promoter strength and the abundance of viral RNA species in RVFV-infected cells. Altogether, these results show that RVFV minigenomes are powerful tools to study transcription and replication and constitute a valuable basis to rescue infectious virus from cDNAs.
裂谷热病毒(RVFV)(白蛉病毒属,布尼亚病毒科)拥有一个由三个负链RNA分子组成的基因组。每个片段都包含3'和5'非编码区,其末端互补序列形成一个柄状结构。我们发现L、M和S片段的转录-复制是受调控的,并且我们建立了一个表达CAT报告基因的微型基因组拯救系统,以研究非编码区在此过程中的作用。基于L、M和S片段的微型基因组被证明可驱动真正的转录和复制,并表达不同水平的CAT报告基因,这表明非编码序列内存在不同的启动子活性。此外,我们发现相对启动子强度与RVFV感染细胞中病毒RNA种类的丰度之间存在良好的相关性。总之,这些结果表明,RVFV微型基因组是研究转录和复制的有力工具,并且构成了从cDNA拯救感染性病毒的宝贵基础。