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Nat Commun. 2018 Mar 16;9(1):1109. doi: 10.1038/s41467-018-03533-0.

Clathrin-adaptor ratio and membrane tension regulate the flat-to-curved transition of the clathrin coat during endocytosis.

Author information

1
Department of Infectious Diseases, Virology, University Hospital Heidelberg, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany.
2
German Cancer Research Center (DKFZ), Im Neuenheimer Feld 581, 69120, Heidelberg, Germany.
3
BioQuant Center, Im Neuenheimer Feld 267, 69120, Heidelberg, Germany.
4
Institute for Theoretical Physics, Heidelberg University, Philosophenweg 19, 69120, Heidelberg, Germany.
5
National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
6
Institute of Pharmacy and Molecular Biotechnology (IPMB), Department of Bioinformatics and Functional Genomics, Heidelberg University, Im Neuenheimer Feld 267, 69120, Heidelberg, Germany.
7
BioQuant Center, Im Neuenheimer Feld 267, 69120, Heidelberg, Germany. schwarz@thphys.uni-heidelberg.de.
8
Institute for Theoretical Physics, Heidelberg University, Philosophenweg 19, 69120, Heidelberg, Germany. schwarz@thphys.uni-heidelberg.de.
9
Department of Infectious Diseases, Virology, University Hospital Heidelberg, Im Neuenheimer Feld 324, 69120, Heidelberg, Germany. s.boulant@dkfz.de.
10
German Cancer Research Center (DKFZ), Im Neuenheimer Feld 581, 69120, Heidelberg, Germany. s.boulant@dkfz.de.

Abstract

Although essential for many cellular processes, the sequence of structural and molecular events during clathrin-mediated endocytosis remains elusive. While it was long believed that clathrin-coated pits grow with a constant curvature, it was recently suggested that clathrin first assembles to form flat structures that then bend while maintaining a constant surface area. Here, we combine correlative electron and light microscopy and mathematical growth laws to study the ultrastructural rearrangements of the clathrin coat during endocytosis in BSC-1 mammalian cells. We confirm that clathrin coats initially grow flat and demonstrate that curvature begins when around 70% of the final clathrin content is acquired. We find that this transition is marked by a change in the clathrin to clathrin-adaptor protein AP2 ratio and that membrane tension suppresses this transition. Our results support the notion that BSC-1 mammalian cells dynamically regulate the flat-to-curved transition in clathrin-mediated endocytosis by both biochemical and mechanical factors.

PMID:
29549258
PMCID:
PMC5856840
DOI:
10.1038/s41467-018-03533-0
[Indexed for MEDLINE]
Free PMC Article

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