Induced Nanoscale Membrane Curvature Bypasses the Essential Endocytic Function of Clathrin

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2022 May 9

PMID 35532382

Authors

Robert C Cail

Cyna R Shirazinejad

David G Drubin

Affiliations

Soon will be listed here.

Abstract

During clathrin-mediated endocytosis (CME), flat plasma membrane is remodeled to produce nanometer-scale vesicles. The mechanisms underlying this remodeling are not completely understood. The ability of clathrin to bind membranes of distinct geometries casts uncertainty on its specific role in curvature generation/stabilization. Here, we used nanopatterning to produce substrates for live-cell imaging, with U-shaped features that bend the ventral plasma membrane of a cell into shapes resembling energetically unfavorable CME intermediates. This induced membrane curvature recruits CME proteins, promoting endocytosis. Upon AP2, FCHo1/2, or clathrin knockdown, CME on flat substrates is severely diminished. However, induced membrane curvature recruits CME proteins in the absence of FCHo1/2 or clathrin and rescues CME dynamics/cargo uptake after clathrin (but not AP2 or FCHo1/2) knockdown. Induced membrane curvature enhances CME protein recruitment upon branched actin assembly inhibition under elevated membrane tension. These data establish that membrane curvature assists in CME nucleation and that the essential function of clathrin during CME is to facilitate curvature evolution, rather than scaffold protein recruitment.

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References

Liu Z, Tan J, Cohen D, Yang M, Sniadecki N, Alom Ruiz S . Mechanical tugging force regulates the size of cell-cell junctions. Proc Natl Acad Sci U S A. 2010; 107(22):9944-9. PMC: 2890446. DOI: 10.1073/pnas.0914547107. View

Dannhauser P, Ungewickell E . Reconstitution of clathrin-coated bud and vesicle formation with minimal components. Nat Cell Biol. 2012; 14(6):634-9. DOI: 10.1038/ncb2478. View

Taylor M, Perrais D, Merrifield C . A high precision survey of the molecular dynamics of mammalian clathrin-mediated endocytosis. PLoS Biol. 2011; 9(3):e1000604. PMC: 3062526. DOI: 10.1371/journal.pbio.1000604. View

Liu J, Sun Y, Drubin D, Oster G . The mechanochemistry of endocytosis. PLoS Biol. 2009; 7(9):e1000204. PMC: 2742711. DOI: 10.1371/journal.pbio.1000204. View

Yarar D, Waterman-Storer C, Schmid S . A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis. Mol Biol Cell. 2004; 16(2):964-75. PMC: 545926. DOI: 10.1091/mbc.e04-09-0774. View

Daboussi L, Costaguta G, Payne G . Phosphoinositide-mediated clathrin adaptor progression at the trans-Golgi network. Nat Cell Biol. 2012; 14(3):239-48. PMC: 4855891. DOI: 10.1038/ncb2427. View

Ford M, Mills I, Peter B, Vallis Y, Praefcke G, Evans P . Curvature of clathrin-coated pits driven by epsin. Nature. 2002; 419(6905):361-6. DOI: 10.1038/nature01020. View

Ma L, Umasankar P, Wrobel A, Lymar A, McCoy A, Holkar S . Transient Fcho1/2⋅Eps15/R⋅AP-2 Nanoclusters Prime the AP-2 Clathrin Adaptor for Cargo Binding. Dev Cell. 2016; 37(5):428-43. PMC: 4921775. DOI: 10.1016/j.devcel.2016.05.003. View

Kazazic M, Bertelsen V, Pedersen K, Vuong T, Grandal M, Rodland M . Epsin 1 is involved in recruitment of ubiquitinated EGF receptors into clathrin-coated pits. Traffic. 2008; 10(2):235-45. DOI: 10.1111/j.1600-0854.2008.00858.x. View

10.

Tinevez J, Perry N, Schindelin J, Hoopes G, Reynolds G, Laplantine E . TrackMate: An open and extensible platform for single-particle tracking. Methods. 2016; 115:80-90. DOI: 10.1016/j.ymeth.2016.09.016. View

11.

Doyon J, Zeitler B, Cheng J, Cheng A, Cherone J, Santiago Y . Rapid and efficient clathrin-mediated endocytosis revealed in genome-edited mammalian cells. Nat Cell Biol. 2011; 13(3):331-7. PMC: 4113319. DOI: 10.1038/ncb2175. View

12.

Sokac A, Co C, Taunton J, Bement W . Cdc42-dependent actin polymerization during compensatory endocytosis in Xenopus eggs. Nat Cell Biol. 2003; 5(8):727-32. DOI: 10.1038/ncb1025. View

13.

Sochacki K, Taraska J . From Flat to Curved Clathrin: Controlling a Plastic Ratchet. Trends Cell Biol. 2019; 29(3):241-256. DOI: 10.1016/j.tcb.2018.12.002. View

14.

Tan X, Luo M, Liu A . Clathrin-mediated endocytosis regulates fMLP-mediated neutrophil polarization. Heliyon. 2018; 4(9):e00819. PMC: 6157066. DOI: 10.1016/j.heliyon.2018.e00819. View

15.

Avinoam O, Schorb M, Beese C, Briggs J, Kaksonen M . ENDOCYTOSIS. Endocytic sites mature by continuous bending and remodeling of the clathrin coat. Science. 2015; 348(6241):1369-72. DOI: 10.1126/science.aaa9555. View

16.

Merrifield C, Feldman M, Wan L, Almers W . Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits. Nat Cell Biol. 2002; 4(9):691-8. DOI: 10.1038/ncb837. View

17.

McMahon H, Boucrot E . Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol. 2011; 12(8):517-33. DOI: 10.1038/nrm3151. View

18.

Chen C, Mrksich M, Huang S, Whitesides G, Ingber D . Geometric control of cell life and death. Science. 1997; 276(5317):1425-8. DOI: 10.1126/science.276.5317.1425. View

19.

Hong S, Cortesio C, Drubin D . Machine-Learning-Based Analysis in Genome-Edited Cells Reveals the Efficiency of Clathrin-Mediated Endocytosis. Cell Rep. 2015; 12(12):2121-30. PMC: 4610353. DOI: 10.1016/j.celrep.2015.08.048. View

20.

Zhao W, Hanson L, Lou H, Akamatsu M, Chowdary P, Santoro F . Nanoscale manipulation of membrane curvature for probing endocytosis in live cells. Nat Nanotechnol. 2017; 12(8):750-756. PMC: 5544585. DOI: 10.1038/nnano.2017.98. View