Surface Immobilization of Viruses and Nanoparticles Elucidates Early Events in Clathrin-Mediated Endocytosis,ACS Infectious Diseases

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Surface Immobilization of Viruses and Nanoparticles Elucidates Early Events in Clathrin-Mediated Endocytosis
ACS Infectious Diseases ( IF 4.0 ) Pub Date : 2018-09-10 00:00:00 , DOI: 10.1021/acsinfecdis.8b00134
Marta Fratini _{1,

2,

3} , Tina Wiegand _{2,

3} , Charlotta Funaya ₄ , Zhongxiang Jiang ₅ , Pranav N. M. Shah ₁ , Joachim P. Spatz _{2,

3} , Elisabetta Ada Cavalcanti-Adam _{2,

3} , Steeve Boulant ₁

Affiliation

Clathrin-mediated endocytosis (CME) is an important entry pathway for viruses. Here, we applied click chemistry to covalently immobilize reovirus on surfaces to study CME during early host–pathogen interactions. To uncouple chemical and physical properties of viruses and determine their impact on CME initiation, we used the same strategy to covalently immobilize nanoparticles of different sizes. Using fluorescence live microscopy and electron microscopy, we confirmed that clathrin recruitment depends on particle size and discovered that the maturation into clathrin-coated vesicles (CCVs) is independent from cargo internalization. Surprisingly, we found that the final size of CCVs appears to be imprinted on the clathrin coat at early stages of cargo–cell interactions. Our approach has allowed us to unravel novel aspects of early interactions between viruses and the clathrin machinery that influence late stages of CME and CCVs formation. This method can be easily and broadly applied to the field of nanotechnology, endocytosis, and virology.

中文翻译：

病毒和纳米粒子的表面固定阐明了网格蛋白介导的内吞作用的早期事件。

网格蛋白介导的内吞作用（CME）是病毒的重要进入途径。在这里，我们应用点击化学技术将呼肠孤病毒共价固定在表面上，以研究早期宿主与病原体相互作用期间的CME。为了解耦病毒的化学和物理特性并确定其对CME引发的影响，我们使用了相同的策略来共价固定不同大小的纳米粒子。使用荧光实时显微镜和电子显微镜，我们确认网格蛋白的募集取决于颗粒大小，并发现网格蛋白涂层的囊泡（CCV）的成熟与货物内在化无关。出乎意料的是，我们发现CCV的最终尺寸似乎在货物-细胞相互作用的早期被印在网格蛋白涂层上。我们的方法使我们能够揭示影响CME和CCV形成后期的病毒与网格蛋白机制之间早期相互作用的新方面。这种方法可以轻松，广泛地应用于纳米技术，内吞作用和病毒学领域。

更新日期：2018-09-10

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