Gamarnik A V, Böddeker N, Andino R
Department of Microbiology and Immunology, University of California, San Francisco, California 94143-0414, USA.
J Virol. 2000 Dec;74(24):11983-7. doi: 10.1128/jvi.74.24.11983-11987.2000.
We have previously shown that Xenopus oocytes require coinjection of both poliovirus RNA and HeLa cell extracts to support a complete cycle of viral replication yielding high levels of infectious viral particles. This novel system provides a tool for identifying host factors and for biochemically dissect individual steps that lead to virus production. Here we demonstrate that Xenopus oocytes are able to support replication of other picornaviruses such as human rhinovirus 14 and mengovirus. Unlike poliovirus, microinjection of mengovirus RNA yields high viral titers (about 10(7) PFU/oocyte) without the need for coinjection of additional cell extracts. In contrast, formation of infectious rhinovirus particles requires coinjection of human cell extracts. We found that one of these human factors is required for efficient rhinovirus translation. Our findings uncover differences in the host factor requirements among members of the picornavirus family and provide the means to identify the human protein(s) involved in rhinovirus production.
我们之前已经表明,非洲爪蟾卵母细胞需要同时注射脊髓灰质炎病毒RNA和HeLa细胞提取物,才能支持病毒复制的完整周期,从而产生高水平的感染性病毒颗粒。这个新系统为识别宿主因子以及从生化角度剖析导致病毒产生的各个步骤提供了一个工具。在此,我们证明非洲爪蟾卵母细胞能够支持其他小RNA病毒的复制,如人鼻病毒14型和脑心肌炎病毒。与脊髓灰质炎病毒不同,显微注射脑心肌炎病毒RNA可产生高病毒滴度(约10⁷ PFU/卵母细胞),而无需同时注射额外的细胞提取物。相比之下,感染性鼻病毒颗粒的形成需要同时注射人细胞提取物。我们发现这些人类因子之一是鼻病毒有效翻译所必需的。我们的研究结果揭示了小RNA病毒家族成员之间在宿主因子需求上的差异,并提供了识别参与鼻病毒产生的人类蛋白质的方法。