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

Bucher, Delia; Frey, Felix J.; Sochacki, Kem A.; Kummer, Susann; Bergeest, Jan-Philip; Godinez, William J.; Kräusslich, Hans‐Georg; Rohr, Karl; Taraska, Justin W.; Schwarz, Ulrich S.; Boulant, Steeve

doi:10.1038/s41467-018-03533-0

Cited by 119 publications

(178 citation statements)

References 44 publications

(77 reference statements)

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“…In line with this, AP2 has been demonstrated to be crucial for stabilizing nascent clathrin‐coated pits and enhances CCV maturation efficiency through cargo concentration . Furthermore, it has recently been shown that the ratio between AP2 and clathrin changes over the time course of CCV formation, where the transition from a flat into a curved clathrin lattice occurs at the point when incorporation of additional AP2 into the coat reaches a plateau . In conjunction with this observation, our findings would suggest that AP2 incorporation increases coat stiffness to a level that exceeds the stiffness of the membrane in order to support the initiation of membrane curvature.…”

Section: Resultssupporting

confidence: 84%

The AP2 adaptor enhances clathrin coat stiffness

et al. 2019

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Deformation of the plasma membrane into clathrin‐coated vesicles is a critical step in clathrin‐mediated endocytosis and requires the orchestrated assembly of clathrin and endocytic adaptors into a membrane‐associated protein coat. The individual role of these membrane‐bending and curvature‐stabilizing factors is subject to current debate. As such, it is unclear whether the clathrin coat itself is stiff enough to impose curvature and if so, whether this could be effectively transferred to the membrane by the linking adaptor proteins. We have recently demonstrated that clathrin alone is sufficient to form membrane buds in vitro. Here, we use atomic force microscopy to assess the contributions of clathrin and its membrane adaptor protein 2 (AP2) to clathrin coat stiffness, which determines the mechanics of vesicle formation. We found that clathrin coats are less than 10‐fold stiffer than the membrane they enclose, suggesting a delicate balance between the forces harnessed from clathrin coat formation and those required for membrane bending. We observed that clathrin adaptor protein AP2 increased the stiffness of coats formed from native clathrin, but did not affect less‐flexible coats formed from clathrin lacking the light chain subunits. We thus propose that clathrin light chains are important for clathrin coat flexibility and that AP2 facilitates efficient cargo sequestration during coated vesicle formation by modulating clathrin coat stiffness.

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Section: Resultssupporting

confidence: 84%

The AP2 adaptor enhances clathrin coat stiffness

et al. 2019

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“…This value is close to the required membrane bending energy so clathrin cage formation might contribute to membrane bending for low membrane tensions. However, the observation of flat clathrin lattices in cells suggests that the polymerization energy does not directly lead to membrane curvature without contributions of other membrane‐deforming mechanisms . Thus, clathrin may contribute to—though not dominate—membrane bending in vivo .…”

Section: Membrane‐binding Proteins and Lipid‐mediated Mechanismsmentioning

confidence: 99%

Molecular mechanisms of force production in clathrin‐mediated endocytosis

Lacy

Ravindra

et al. 2018

FEBS Letters

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During clathrin-mediated endocytosis (CME), a flat patch of membrane is invaginated and pinched off to release a vesicle into the cytoplasm. In yeast CME, over 60 proteins—including a dynamic actin meshwork—self-assemble to deform the plasma membrane. Several models have been proposed for how actin and other molecules produce the forces necessary to overcome the mechanical barriers of membrane tension and turgor pressure, but the precise mechanisms and a full picture of their interplay are still not clear. In this review, we discuss the evidence for these force production models from a quantitative perspective and propose future directions for experimental and theoretical work that could clarify their various contributions.

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“…Thus, we conclude that through controlling CHC's conformational flexibility, CLCs reduce excessive interactions between triskelia and promote lattice rearrangement. Weakening interactions between triskelia further explains how CLCs raise the critical assembly concentration for clathrin 41,75 and create dynamic lattices amenable to regulation by adaptor proteins 34,76 .…”

Section: Discussionmentioning

confidence: 99%

Clathrin light chain diversity regulates membrane deformation in vitro and synaptic vesicle formation in vivo

Redlingshoefer

McLeod

Camus

et al. 2019

Preprint

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Clathrin light chain (CLC) subunits in vertebrates are encoded by paralogous genes CLTA and CLTB and both gene products undergo alternative splicing in neurons. To understand how this CLC diversity influences neuronal clathrin function, we characterised the biophysical properties of clathrin comprising individual CLC variants for correlation with neuronal phenotypes of mice lacking either CLC-encoding gene. CLC variants differentially influenced clathrin knee conformation within assemblies, and clathrin lattices with neuronal CLC mixtures were more effective in membrane bending than those with single neuronal isoforms nCLCa or nCLCb. Correspondingly, electrophysiological recordings revealed that neurons from mice deficient for nCLCa or nCLCb were defective in synaptic vesicle recycling. Mice with only nCLCb had a reduced synaptic vesicle pool compared to wild-type mice, while nCLCa-only mice had increased synaptic vesicle numbers. These findings highlight functional differences between the CLC isoforms and show that isoform mixing influences tissue-specific clathrin function in neurons.

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

Cited by 119 publications

References 44 publications

The AP2 adaptor enhances clathrin coat stiffness

The AP2 adaptor enhances clathrin coat stiffness

Molecular mechanisms of force production in clathrin‐mediated endocytosis

Clathrin light chain diversity regulates membrane deformation in vitro and synaptic vesicle formation in vivo

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