Genome-wide analysis of influenza viral RNA and nucleoprotein association.
Genome-wide analysis of influenza viral RNA and nucleoprotein association.
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DOI：
10.1093/nar/gkx584
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发表时间：
2017-09-06
影响因子：
14.9
通讯作者：
Lakdawala SS
中科院分区：
文献类型：
--
作者：
Lee N;Le Sage V;Nanni AV;Snyder DJ;Cooper VS;Lakdawala SS
甲型流感病毒（IAV）基因组由八个单链RNA片段组成，这些片段被病毒核蛋白（NP）分子包裹。传统上，NP和病毒RNA （vRNA）之间的相互作用被描述为“串珠”的统一模式。利用交联免疫沉淀（HITS-CLIP）分离RNA的高通量测序，我们鉴定了两种H1N1 IAV毒株在病毒粒子中NP的vRNA结合谱。与目前流行的vRNA包装模式相反，在A/WSN/1933和A/California/07/2009菌株中，NP并不均匀地结合vRNA，相反，每个vRNA片段都表现出独特的结合特征，包含丰富或缺乏NP关联的位点。有趣的是，尽管有广泛的序列守恒，但这两种H1N1病毒株具有相似但不同的NP结合谱。基于对DNA反义寡核苷酸介导的RNase H酶切不敏感，通过HITS-CLIP鉴定的峰被验证为真正的NP结合位点。此外，对NP峰的核苷酸含量分析显示，与全基因组核苷酸含量相比，这些位点相对富含g和缺乏u，表明体内NP关联存在尚未确定的序列偏差。总之，我们对NP-vRNA相互作用的全基因组研究对理解流感vRNA结构和基因组包装具有重要意义。
Influenza A virus (IAV) genomes are composed of eight single-stranded RNA segments that are coated by viral nucleoprotein (NP) molecules. Classically, the interaction between NP and viral RNA (vRNA) is depicted as a uniform pattern of ‘beads on a string’. Using high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP), we identified the vRNA binding profiles of NP for two H1N1 IAV strains in virions. Contrary to the prevailing model for vRNA packaging, NP does not bind vRNA uniformly in the A/WSN/1933 and A/California/07/2009 strains, but instead each vRNA segment exhibits a unique binding profile, containing sites that are enriched or poor in NP association. Intriguingly, both H1N1 strains have similar yet distinct NP binding profiles despite extensive sequence conservation. Peaks identified by HITS-CLIP were verified as true NP binding sites based on insensitivity to DNA antisense oligonucleotide-mediated RNase H digestion. Moreover, nucleotide content analysis of NP peaks revealed that these sites are relatively G-rich and U-poor compared to the genome-wide nucleotide content, indicating an as-yet unidentified sequence bias for NP association in vivo. Taken together, our genome-wide study of NP–vRNA interaction has implications for the understanding of influenza vRNA architecture and genome packaging.
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影响因子：
14.8
作者：
通讯作者：
--
影响因子：
4.6
作者：
Liu CL;Hung HC;Lo SC;Chiang CH;Chen IJ;Hsu JT;Hou MH
通讯作者：
Hou MH
DOI：
10.1073/pnas.73.9.3045
发表时间：
1976-01-01
影响因子：
11.1
作者：
MCGEOCH, D;FELLNER, P;NEWTON, C
通讯作者：
NEWTON, C
影响因子：
2.7
作者：
HATADA, E;HASEGAWA, M;FUKUDA, R
通讯作者：
FUKUDA, R
DOI：
10.1038/nrmicro3367
发表时间：
2015-01
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
Eisfeld AJ;Neumann G;Kawaoka Y
通讯作者：
Kawaoka Y