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ACS Infect Dis. 2018 Nov 9;4(11):1585-1600. doi: 10.1021/acsinfecdis.8b00134. Epub 2018 Sep 26.

Surface Immobilization of Viruses and Nanoparticles Elucidates Early Events in Clathrin-Mediated Endocytosis.

Author information

1
Heidelberg University , Department of Infectious Diseases, Virology and German Cancer Research Center , Im Neuenheimer Feld 581 , 69120 Heidelberg , Germany.
2
Max Planck Institute for Medical Research , Department of Cellular Biophysics , Jahnstrasse 29 , 69120 Heidelberg , Germany.
3
Heidelberg University , Institute of Physical Chemistry, Department of Biophysical Chemistry , Im Neuenheimer Feld 253 , 69120 , Heidelberg , Germany.
4
Heidelberg University , Electron Microscopy Core Facility , Im Neuenheimer Feld 345 , 69120 Heidelberg , Germany.
5
Leica Microsystems GmbH , Am Friedensplatz 3 , 68165 Mannheim , Germany.

Abstract

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.

KEYWORDS:

clathrin-mediated endocytosis; click chemistry; endocytosis; membrane curvature; nanoparticles; viruses

PMID:
30200751
DOI:
10.1021/acsinfecdis.8b00134
[Indexed for MEDLINE]
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