《Nature》H1N1病毒鸠占鹊巢

　　

生物通报道，来自Burnham医学研究所，Salk研究所等处的科学家关于甲流H1N1病毒研究取得新进展，相关成果文章Human host factors required for influenza virus replication发表在Nature在线版上。

 

文章通讯作者是来自西奈山医学院的Megan L. Shaw助理教授与Burnham医学研究所所的首席研究员sumit k chanda副教授。

 

流感病毒，是一种造成人类及动物患流行性感冒的RNA病毒，编码11个蛋白。病毒进入宿主细胞后会利用宿主细胞的遗传物质和细胞器进行自我复制。

 

病毒侵入体内后依靠血凝素吸附于宿主细胞表面，经过吞饮进入胞浆；进入胞浆之后病毒包膜与细胞膜融合释放出包含的ss-RNA；ss-RNA的八个节段在胞浆内编码RNA多聚酶、核蛋白、基质蛋白、膜蛋白、血凝素、神经氨酸酶、非结构蛋白等构件；基质蛋白、膜蛋白、血凝素、神经氨酸酶等编码蛋白在内质网或高尔基体上组装M蛋白和包膜；在细胞核内，病毒的遗传物质不断复制并与核蛋白、RNA多聚酶等组建病毒核心；最终病毒核心与膜上的M蛋白和包膜结合，经过出芽释放到细胞之外，复制的周期大约8个小时。

 

病毒复制的过程所必须的物质都来自宿主细胞，了解宿主细胞在病毒复制过程中所发挥的作用有助于科学家研究开发抗甲流病毒药物。

 

在本研究中，科学家们应用一个建立在RNAi技术上的复合研究体系对宿主细胞参与病毒复制的因子进行扫描，结果发现在病毒复制早期，存在295种细胞辅助因子帮助病毒复制。

 

深入研究发现，这些辅助因子参与激酶信号通路，泛素化，磷酸化活动，其中181个辅助因子组成宿主-病毒调节网路。

 

295个辅助因子中有219个被确认为甲流野生毒株生长的必须因子，其中，23个因子是甲流病毒进入宿主细胞的关键调控因子，它们分别涉及ATP酶活性，糖原合成酶等多种重要酶的活性调控。

 

研究还发现10种蛋白在病毒侵入宿主细胞后的复制过程具有重要的意义。

 

显然，甲流病毒必须依赖宿主细胞才能完成复制工作，因此，科学家们可以通过以宿主细胞中的某些因子为靶位制备抗病毒复制的药物。

 

研究小组发现了一个小分子抑制物可抑制好几个宿主活性因子，这样便可防止甲流病毒复制。

（生物通 小茜）

生物通推荐原文检索

Human host factors required for influenza virus replicationnear-final version

Renate König1,9, Silke Stertz4,9, Yingyao Zhou7, Atsushi Inoue1, H. -Heinrich Hoffmann4, Suchita Bhattacharyya2, Judith G. Alamares4, Donna M. Tscherne4, Mila B. Ortigoza4, Yuhong Liang4, Qinshan Gao4, Shane E. Andrews3, Sourav Bandyopadhyay8, Paul De Jesus1, Buu P. Tu7, Lars Pache1, Crystal Shih1, Anthony Orth7, Ghislain Bonamy7, Loren Miraglia7, Trey Ideker8, Adolfo García-Sastre4,5,6, John A. T. Young2, Peter Palese4,5, Megan L. Shaw4,9 & Sumit K. Chanda1,9

 

Infectious and Inflammatory Disease Center, Burnham Institute for Medical Research, 10901 North Torrey Pines Road,

Nomis Center for Immunobiology and Microbial Pathogenesis, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road,

Gene Expression Laboratory, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA

Department of Microbiology,

Department of Medicine, Division of Infectious Diseases,

Global Health and Emerging Pathogens Institute, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, New York 10029, USA

Genomics Institute of the Novartis Research Foundation, 10675 John J Hopkins Drive, San Diego, California 92121, USA

Departments of Medicine and Bioengineering, University of California, San Diego, Pharmaceutical Sciences Building, 9500 Gilman Drive, La Jolla, California 92093, USA

These authors contributed equally to this work.

Correspondence to: Megan L. Shaw4,9Sumit K. Chanda1,9 Correspondence and requests for materials should be addressed to S.K.C. (Email: [e-mail]) or M.L.S. (Email: [e-mail]).

 

【Abstract】

Influenza A virus is an RNA virus that encodes up to 11 proteins and this small coding capacity demands that the virus use the host cellular machinery for many aspects of its life cycle1. Knowledge of these host cell requirements not only informs us of the molecular pathways exploited by the virus but also provides further targets that could be pursued for antiviral drug development. Here we use an integrative systems approach, based on genome-wide RNA interference screening, to identify 295 cellular cofactors required for early-stage influenza virus replication. Within this group, those involved in kinase-regulated signalling, ubiquitination and phosphatase activity are the most highly enriched, and 181 factors assemble into a highly significant host–pathogen interaction network. Moreover, 219 of the 295 factors were confirmed to be required for efficient wild-type influenza virus growth, and further analysis of a subset of genes showed 23 factors necessary for viral entry, including members of the vacuolar ATPase (vATPase) and COPI-protein families, fibroblast growth factor receptor (FGFR) proteins, and glycogen synthase kinase 3 (GSK3)-β. Furthermore, 10 proteins were confirmed to be involved in post-entry steps of influenza virus replication. These include nuclear import components, proteases, and the calcium/calmodulin-dependent protein kinase (CaM kinase) IIβ (CAMK2B). Notably, growth of swine-origin H1N1 influenza virus is also dependent on the identified host factors, and we show that small molecule inhibitors of several factors, including vATPase and CAMK2B, antagonize influenza virus replication.