Force Redistribution in Clathrin-mediated Endocytosis Revealed by Coiled-coil Force Sensors

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2023 Oct 13

PMID 37831774

Authors

Yuan Ren

Jie Yang

Barbara Fujita

Huaizhou Jin

Yongli Zhang

Julien Berro

Affiliations

Soon will be listed here.

Abstract

Forces are central to countless cellular processes, yet in vivo force measurement at the molecular scale remains difficult if not impossible. During clathrin-mediated endocytosis, forces produced by the actin cytoskeleton are transmitted to the plasma membrane by a multiprotein coat for membrane deformation. However, the magnitudes of these forces remain unknown. Here, we present new in vivo force sensors that induce protein condensation under force. We measured the forces on the fission yeast Huntingtin-Interacting Protein 1 Related (HIP1R) homolog End4p, a protein that links the membrane to the actin cytoskeleton. End4p is under ~19-piconewton force near the actin cytoskeleton, ~11 piconewtons near the clathrin lattice, and ~9 piconewtons near the plasma membrane. Our results demonstrate that forces are collected and redistributed across the endocytic machinery.

Citing Articles

Genetically encoded mechano-sensors with versatile readouts and compact size.

Ren Y, Yang J, Saito T, Glomb O, Mousavi S, Naughton B bioRxiv. 2025; .

PMID: 39868242 PMC: 11760715. DOI: 10.1101/2025.01.16.633409.

Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals.

Gest A, Sahan A, Zhong Y, Lin W, Mehta S, Zhang J Chem Rev. 2024; 124(22):12573-12660.

PMID: 39535501 PMC: 11613326. DOI: 10.1021/acs.chemrev.4c00293.

Fast Fission Yeast Genome Editing by CRISPR/Cas9 Using Gap Repair and Fluoride Selection.

Ren Y, Fernandez R, Saito T, Fujita B, Mousavi I, Berro J Methods Mol Biol. 2024; 2862:141-154.

PMID: 39527198 DOI: 10.1007/978-1-0716-4168-2_10.

Cross-regulations of two connected domains form a mechanical circuit for steady force transmission during clathrin-mediated endocytosis.

Ren Y, Yang J, Fujita B, Zhang Y, Berro J Cell Rep. 2024; 43(9):114725.

PMID: 39276354 PMC: 11476202. DOI: 10.1016/j.celrep.2024.114725.

A Tiny Viral Protein, SARS-CoV-2-ORF7b: Functional Molecular Mechanisms.

Mansueto G, Fusco G, Colonna G Biomolecules. 2024; 14(5).

PMID: 38785948 PMC: 11118181. DOI: 10.3390/biom14050541.

References

Kozak M, Kaksonen M . Condensation of Ede1 promotes the initiation of endocytosis. Elife. 2022; 11. PMC: 9064294. DOI: 10.7554/eLife.72865. View

Mettlen M, Chen P, Srinivasan S, Danuser G, Schmid S . Regulation of Clathrin-Mediated Endocytosis. Annu Rev Biochem. 2018; 87:871-896. PMC: 6383209. DOI: 10.1146/annurev-biochem-062917-012644. View

Basu R, Munteanu E, Chang F . Role of turgor pressure in endocytosis in fission yeast. Mol Biol Cell. 2014; 25(5):679-87. PMC: 3937093. DOI: 10.1091/mbc.E13-10-0618. View

Truebestein L, Leonard T . Coiled-coils: The long and short of it. Bioessays. 2016; 38(9):903-16. PMC: 5082667. DOI: 10.1002/bies.201600062. View

Grashoff C, Hoffman B, Brenner M, Zhou R, Parsons M, Yang M . Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature. 2010; 466(7303):263-6. PMC: 2901888. DOI: 10.1038/nature09198. View

Wang X, Ha T . Defining single molecular forces required to activate integrin and notch signaling. Science. 2013; 340(6135):991-4. PMC: 3710701. DOI: 10.1126/science.1231041. View

Warren R, Kessels M, Keen J, Heuser J, Drubin D . The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro. J Cell Biol. 2001; 154(6):1209-23. PMC: 2150824. DOI: 10.1083/jcb.200106089. View

Lemiere J, Ren Y, Berro J . Rapid adaptation of endocytosis, exocytosis, and eisosomes after an acute increase in membrane tension in yeast cells. Elife. 2021; 10. PMC: 9045820. DOI: 10.7554/eLife.62084. View

Goktas M, Luo C, Sullan R, Bergues-Pupo A, Lipowsky R, Vila Verde A . Molecular mechanics of coiled coils loaded in the shear geometry. Chem Sci. 2018; 9(20):4610-4621. PMC: 5969510. DOI: 10.1039/c8sc01037d. View

10.

Fujiwara I, Zweifel M, Courtemanche N, Pollard T . Latrunculin A Accelerates Actin Filament Depolymerization in Addition to Sequestering Actin Monomers. Curr Biol. 2018; 28(19):3183-3192.e2. PMC: 6179359. DOI: 10.1016/j.cub.2018.07.082. View

11.

Skruzny M, Brach T, Ciuffa R, Rybina S, Wachsmuth M, Kaksonen M . Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis. Proc Natl Acad Sci U S A. 2012; 109(38):E2533-42. PMC: 3458359. DOI: 10.1073/pnas.1207011109. View

12.

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

13.

Bracha D, Walls M, Wei M, Zhu L, Kurian M, Avalos J . Mapping Local and Global Liquid Phase Behavior in Living Cells Using Photo-Oligomerizable Seeds. Cell. 2018; 175(6):1467-1480.e13. PMC: 6724719. DOI: 10.1016/j.cell.2018.10.048. View

14.

Pandya M, SPOONER G, Sunde M, Thorpe J, Rodger A, Woolfson D . Sticky-end assembly of a designed peptide fiber provides insight into protein fibrillogenesis. Biochemistry. 2000; 39(30):8728-34. DOI: 10.1021/bi000246g. View

15.

Abella M, Andruck L, Malengo G, Skruzny M . Actin-generated force applied during endocytosis measured by Sla2-based FRET tension sensors. Dev Cell. 2021; 56(17):2419-2426.e4. DOI: 10.1016/j.devcel.2021.08.007. View

16.

Tang H, Xu J, Cai M . Pan1p, End3p, and S1a1p, three yeast proteins required for normal cortical actin cytoskeleton organization, associate with each other and play essential roles in cell wall morphogenesis. Mol Cell Biol. 1999; 20(1):12-25. PMC: 85029. DOI: 10.1128/MCB.20.1.12-25.2000. View

17.

Sun Y, Schoneberg J, Chen X, Jiang T, Kaplan C, Xu K . Direct comparison of clathrin-mediated endocytosis in budding and fission yeast reveals conserved and evolvable features. Elife. 2019; 8. PMC: 6908435. DOI: 10.7554/eLife.50749. View

18.

Skruzny M, Desfosses A, Prinz S, Dodonova S, Gieras A, Uetrecht C . An organized co-assembly of clathrin adaptors is essential for endocytosis. Dev Cell. 2015; 33(2):150-62. DOI: 10.1016/j.devcel.2015.02.023. View

19.

Bracha D, Walls M, Brangwynne C . Probing and engineering liquid-phase organelles. Nat Biotechnol. 2019; 37(12):1435-1445. DOI: 10.1038/s41587-019-0341-6. View

20.

Kukulski W, Schorb M, Kaksonen M, Briggs J . Plasma membrane reshaping during endocytosis is revealed by time-resolved electron tomography. Cell. 2012; 150(3):508-20. DOI: 10.1016/j.cell.2012.05.046. View