Groseth Allison, Feldmann Heinz, Theriault Steven, Mehmetoglu Gülsah, Flick Ramon
National Laboratory for Zoonotic Diseases and Special Pathogens, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB R3E 3R2, Canada.
J Virol. 2005 Apr;79(7):4425-33. doi: 10.1128/JVI.79.7.4425-4433.2005.
In general, Ebola viruses are well known for their ability to cause severe hemorrhagic fever in both human and nonhuman primates. However, despite substantial sequence homology to other members of the family Filoviridae, Reston ebolavirus displays reduced pathogenicity for nonhuman primates and has never been demonstrated to cause clinical disease in humans, despite its ability to cause infection. In order to develop a tool to explore potential roles for transcription and replication in the reduced pathogenicity of Reston ebolavirus, we developed an RNA polymerase I (Pol I)-driven minigenome system. Here we demonstrate successful Reston ebolavirus minigenome rescue, including encapsidation, transcription, and replication, as well as the packaging of minigenome transcripts into progeny particles. The Pol I-driven Reston ebolavirus minigenome system provides a higher signal intensity with less background (higher signal-to-noise ratio) than a comparable T7-driven Reston ebolavirus minigenome system which was developed simultaneously. Successful Reston ebolavirus minigenome rescue was also achieved by the use of helper plasmids derived from the closely related Zaire ebolavirus or the more distantly related Lake Victoria marburgvirus. The use of heterologous helper plasmids in the Reston ebolavirus minigenome system yielded levels of reporter expression which far exceeded the level produced by the homologous helper plasmids. This comparison between minigenomes and helper plasmids from different filovirus species and genera indicates that inherent differences in the transcription and/or replication capacities of the ribonucleoprotein complexes of pathogenic and apathogenic filoviruses may exist, as these observations were confirmed in a Lake Victoria marburgvirus minigenome system.
一般来说，埃博拉病毒以其在人类和非人类灵长类动物中引发严重出血热的能力而闻名。然而，尽管与丝状病毒科的其他成员有大量序列同源性，但雷斯顿埃博拉病毒对非人类灵长类动物的致病性较低，并且尽管它有感染能力，但从未被证明会在人类中引起临床疾病。为了开发一种工具来探索转录和复制在雷斯顿埃博拉病毒致病性降低中的潜在作用，我们开发了一种由RNA聚合酶I（Pol I）驱动的微型基因组系统。在这里，我们展示了成功的雷斯顿埃博拉病毒微型基因组拯救，包括衣壳化、转录和复制，以及微型基因组转录本包装到子代颗粒中。与同时开发的类似的由T7驱动的雷斯顿埃博拉病毒微型基因组系统相比，由Pol I驱动的雷斯顿埃博拉病毒微型基因组系统具有更高的信号强度和更低的背景（更高的信噪比）。通过使用来自密切相关的扎伊尔埃博拉病毒或亲缘关系较远的维多利亚湖马尔堡病毒的辅助质粒，也实现了成功的雷斯顿埃博拉病毒微型基因组拯救。在雷斯顿埃博拉病毒微型基因组系统中使用异源辅助质粒产生的报告基因表达水平远远超过同源辅助质粒产生的水平。来自不同丝状病毒物种和属的微型基因组和辅助质粒之间的这种比较表明，致病性和非致病性丝状病毒的核糖核蛋白复合物在转录和/或复制能力上可能存在内在差异，因为这些观察结果在维多利亚湖马尔堡病毒微型基因组系统中得到了证实。
