Mühlberger E, Weik M, Volchkov V E, Klenk H D, Becker S
Institut für Virologie der Philipps-Universität Marburg, 35037 Marburg, Germany.
J Virol. 1999 Mar;73(3):2333-42. doi: 10.1128/JVI.73.3.2333-2342.1999.
The members of the family Filoviridae, Marburg virus (MBGV) and Ebola virus (EBOV), are very similar in terms of morphology, genome organization, and protein composition. To compare the replication and transcription strategies of both viruses, an artificial replication system based on the vaccinia virus T7 expression system was established for EBOV. Specific transcription and replication of an artificial monocistronic minireplicon was demonstrated by reporter gene expression and detection of the transcribed and replicated RNA species. As it was shown previously for MBGV, three of the four EBOV nucleocapsid proteins, NP, VP35, and L, were essential and sufficient for replication. In contrast to MBGV, EBOV-specific transcription was dependent on the presence of the fourth nucleocapsid protein, VP30. When EBOV VP30 was replaced by MBGV VP30, EBOV-specific transcription was observed but with lower efficiency. Exchange of NP, VP35, and L between the two replication systems did not lead to detectable reporter gene expression. It was further observed that neither MBGV nor EBOV were able to replicate the heterologous minigenomes. A chimeric minigenome, however, containing the EBOV leader and the MBGV trailer was encapsidated, replicated, transcribed, and packaged by both viruses.
丝状病毒科成员马尔堡病毒（MBGV）和埃博拉病毒（EBOV）在形态、基因组结构和蛋白质组成方面非常相似。为了比较这两种病毒的复制和转录策略，基于痘苗病毒T7表达系统为埃博拉病毒建立了一个人工复制系统。通过报告基因表达以及对转录和复制的RNA种类的检测，证明了人工单顺反子微型复制子的特异性转录和复制。正如之前对马尔堡病毒所显示的那样，埃博拉病毒的四种核衣壳蛋白中的三种，即核蛋白（NP）、病毒蛋白35（VP35）和依赖RNA的RNA聚合酶（L），对于复制是必不可少且足够的。与马尔堡病毒不同，埃博拉病毒特异性转录依赖于第四种核衣壳蛋白病毒蛋白30（VP30）的存在。当埃博拉病毒的VP30被马尔堡病毒的VP30取代时，可观察到埃博拉病毒特异性转录，但效率较低。两个复制系统之间的NP、VP35和L的交换并未导致可检测到的报告基因表达。进一步观察到，马尔堡病毒和埃博拉病毒都无法复制异源微型基因组。然而，一个包含埃博拉病毒前导序列和马尔堡病毒尾序列的嵌合微型基因组被两种病毒包装、复制、转录和包裹。
