High-throughput super-resolution analysis of influenza virus pleomorphism reveals insights into viral spatial organization.
High-throughput super-resolution analysis of influenza virus pleomorphism reveals insights into viral spatial organization.
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DOI：
10.1371/journal.ppat.1011484
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发表时间：
2023-06
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
许多病毒形成高度多形性的颗粒。在流感中，病毒体结构不仅在病毒组装的背景下令人感兴趣，而且还因为多形性变异可能与感染性和致病性相关。我们使用荧光超分辨率显微镜结合快速自动化分析管道，一种非常适合于研究大量多形性结构的方法，对数千个单独的流感病毒粒子进行成像;获得有关其大小，形态和分布的信息膜嵌入和内部蛋白质。我们观察到广泛的表型变异的细丝大小，和超分辨率图像的傅立叶变换分析表明，没有一般化的共同的空间频率模式的HA或NA的病毒粒子表面上，这表明一个模型的病毒颗粒组装的后代细丝从细胞中的释放发生在一个随机的方式。我们还表明，病毒RNP复合物位于优先Archetti机构内时，观察到这些细丝的末端，这表明这些结构可能在病毒传播中发挥作用。因此，我们的方法提供了令人兴奋的新的见解流感病毒形态，并代表了一个强大的技术，很容易扩展到其他致病病毒的多形性研究。病毒是重要的人类病原体。在流感中，病毒结构和形态与致病性有关;然而，研究病毒结构的常用方法通常通量低，或缺乏解析病毒颗粒特征所需的分辨率。此外，流感病毒细丝由于其脆弱的性质和在病毒传代过程中丝状形态的丧失而经常未被充分研究。为了解决这个问题，我们开发了一种荧光超分辨率显微镜和快速自动化分析管道，可以一次对数千个流感病毒粒子进行成像，获得有关其大小、形态和蛋白质分布的信息。使用这种方法，我们能够表明，有没有特定的交替的表面糖蛋白的细丝，和富含神经氨酸酶的Archetti机构优先房子病毒核糖核蛋白复合物。我们的发现为流感病毒颗粒组装和病毒传播提供了新的见解，此外，我们的多功能方法有可能为病毒研究的多个领域做出贡献。
Many viruses form highly pleomorphic particles. In influenza, virion structure is of interest not only in the context of virus assembly, but also because pleomorphic variations may correlate with infectivity and pathogenicity. We have used fluorescence super-resolution microscopy combined with a rapid automated analysis pipeline, a method well-suited to the study of large numbers of pleomorphic structures, to image many thousands of individual influenza virions; gaining information on their size, morphology and the distribution of membrane-embedded and internal proteins. We observed broad phenotypic variability in filament size, and Fourier transform analysis of super-resolution images demonstrated no generalized common spatial frequency patterning of HA or NA on the virion surface, suggesting a model of virus particle assembly where the release of progeny filaments from cells occurs in a stochastic way. We also showed that viral RNP complexes are located preferentially within Archetti bodies when these were observed at filament ends, suggesting that these structures may play a role in virus transmission. Our approach therefore offers exciting new insights into influenza virus morphology and represents a powerful technique that is easily extendable to the study of pleomorphism in other pathogenic viruses. Viruses are significant human pathogens. In influenza, virus structure and morphology has been linked to pathogenicity; however commonly used methods to study virus structure are often low through-put, or lack the resolution required to resolve virus particle features. In addition, influenza filaments are frequently understudied due to their fragile nature and loss of filamentous morphology during viral passage. In order to address this, we have developed a fluorescence super-resolution microscopy and rapid automated analysis pipeline to image many thousands of individual influenza virions at a time, gaining information on their size, morphology and protein distribution. Using this method we were able to show that there is no specific alternation of the surface glycoproteins in filaments, and that neuraminidase-enriched Archetti bodies preferentially house viral ribonucleoprotein complexes. Our findings offer new insights into influenza virus particle assembly and virus transmission, and further, our versatile methods have the potential to contribute towards multiple areas of virus research.
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DOI：
10.1099/jgv.0.000535
发表时间：
2016-08
期刊：
The Journal of general virology
影响因子：
--
作者：
Dadonaite B;Vijayakrishnan S;Fodor E;Bhella D;Hutchinson EC
通讯作者：
Hutchinson EC
影响因子：
--
作者：
Xiong Y;Kabacoff C;Franca-Koh J;Devreotes PN;Robinson DN;Iglesias PA
通讯作者：
Iglesias PA
影响因子：
6.7
作者：
Lakdawala SS;Lamirande EW;Suguitan AL Jr;Wang W;Santos CP;Vogel L;Matsuoka Y;Lindsley WG;Jin H;Subbarao K
通讯作者：
Subbarao K
影响因子：
7.7
作者：
Laine, Romain F.;Goodfellow, Gemma;Kaminski, Clemens F.
通讯作者：
Kaminski, Clemens F.
影响因子：
64.8
作者：
Ke, Zunlong;Oton, Joaquin;Briggs, John A. G.
通讯作者：
Briggs, John A. G.