A reconstituted replication and transcription system for Ebola virus Reston and comparison with Ebola virus Zaire
A reconstituted replication and transcription system for Ebola virus Reston and comparison with Ebola virus Zaire
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DOI：
10.1016/j.virol.2004.11.018
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发表时间：
2005-02-05
期刊：
影响因子：
3.7
通讯作者：
Mühlberger, E
中科院分区：
文献类型：
--
作者：
Boehmann, Y;Enterlein, S;Mühlberger, E
唯一已知的丝状病毒，据推测对人类没有致病性，是埃博拉病毒（EBOV）莱斯顿。当EBOV Reston和高致病性EBOV Zaire在细胞培养中生长时，复制动力学的比较显示EBOV Reston的明显生长障碍，表明EBOV Reston的复制周期可能被延迟。此外，EBOV Reston引起的病毒引起的细胞病变效应比EBOV Zaire轻得多。为了比较EBOV Reston和Zaire的复制和转录，建立了一个基于反向遗传学的重组EBOV Reston小基因组复制和转录系统。该系统用于交换催化复制和转录的EBOV Zaire和EBOV Reston核衣壳（NC）蛋白NP、VP 35、VP 30和L。此外，嵌合微型基因组的构建含有EBOV扎伊尔的顺式作用复制信号结合的EBOV莱斯顿。令人惊讶的是，顺式作用信号以及几乎所有的NC蛋白可以在EBOV Reston和Zaire之间交换，表明EBOV Zaire和EBOV Reston的复制/转录复合物的高度功能同源性。只有EBOV Zaire VP 35和EBOV Reston L的组合不导致复制和转录活性。虽然这两种蛋白质不构成活性聚合酶复合物，但免疫荧光分析表明它们仍然能够相互作用。(C)2004年爱思唯尔公司All rights reserved.
The only known filovirus, which presumably is not pathogenic for humans, is Ebola virus (EBOV) Reston. When EBOV Reston and the highly pathogenic EBOV Zaire were grown in cell culture, comparison of the replication kinetics showed a clear growth impairment of EBOV Reston, indicating that the replication cycle of EBOV Reston might be delayed. In addition, the cytopathic effect caused by the virus was much milder with EBOV Reston than with EBOV Zaire. To compare replication and transcription of EBOV Reston and Zaire, a reconstituted minigenomic replication and transcription system based on reverse genetics has been established for EBOV Reston. This system was used to exchange the EBOV Zaire and EBOV Reston nucleocapsid (NC) proteins NP, VP35, VP30, and L, which catalyze replication and transcription. Furthermore, chimeric minigenomes were constructed containing the cis-acting replication signals of EBOV Zaire combined with those of EBOV Reston. Surprisingly, the cis-acting signals as well as almost all NC proteins could be exchanged between EBOV Reston and Zaire, suggesting a high degree of functional homology of the replication/transcription complexes of EBOV Zaire and EBOV Reston. Only the combination of EBOV Zaire VP35 and EBOV Reston L did not result in replication and transcription activity. Although these two proteins did not constitute an active polymerase complex, it was shown by immunofluorescence analysis that they were still able to interact. (C) 2004 Elsevier Inc. All rights reserved.