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呼吸道合胞病毒糖蛋白和基因组RNA动态揭示了,在质膜前,微丝的组成

作者: Chiara Zurla1& Philip J. Santangelo

呼吸道合胞病毒糖蛋白和基因组RNA动态揭示了,在质膜前,微丝的组成

人类呼吸道合胞病毒G蛋白在进入和组成丝状病毒中起着重要的作用。在这里,我们报告,在活细胞中,利用大豆凝集素荧光标记,选择性地标记呼吸道合胞病毒G蛋白,而不破坏呼吸道合胞病毒感染性或丝状形态,以及试探呼吸道合胞病毒粒子组合的反应。使用这种方法,我们发现,经过网格蛋白介导的 内噬作用,质膜呼吸道合胞病毒G可以从隔膜迅速回收利用。这一事件后,是动态形式的丝状、分枝的呼吸道合胞病毒颗粒,在再次插入质膜前,与基因组核糖核蛋白和小窝囊泡聚集一起。我们证明,通过破坏微管、抑制分子马达,可以使这一过程终止。总的来说,我们的研究结果表明,在插入到质膜之前,带着基因组RNA呼吸合胞病毒组成、生产病毒丝和装载。

SBA selectively binds RSV G.

a Vero cells infected (top) or mock-infected (bottom) for 24 h with RSV A2 at an MOI of 1 were labeled with SBA-488 at 4 °C (green) before being fixed and stained for RSV G (red), RSV F (blue), and nuclei (gray). Extended focus image is shown. Scale bar represents 10 µm. b Enlarged cropped images from the white boxed regions from part a with intensity profiles showing colocalization along the direction of the white arrow. Scale bar represents 1 µm. c Cells transfected for 24 h with RSV G (top) and RSV F (bottom) mRNA were labeled with SBA-488 at 4 °C (green) before being fixed and stained without permeabilization for RSV G or RSV F (red) and nuclei (gray). Extended focus image is shown. Scale bar represents 15 µm. d Mander’s colocalization coefficient and Pearson’s correlation between RSV G or RSV F and SBA. Dotted line indicates 0 correlation. n = 16 cells per group. Error bars represent 95% confidence interval. Asterisks indicates p < 0.0001 by Mann–Whitney U Test. e Enlarged cropped images from the white boxed regions from part c with YZ cross-sections (left). Intensity profiles, indicated by the white line on the YZ cross-section, show the expressed RSV protein on a higher plane than the nucleus as indicated by the arrow. Extended focus cropped images are shown


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http://www.readcube.com/articles/10.1038/s41467-017-00732-z


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