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Coupling between Clathrin-Coated-Pit Invagination, Cortactin Recruitment, and Membrane Scission Observed in Live Cells
Open accessAbstract
During clathrin-mediated endocytosis, membrane scission marks the isolation of a cargo-laden clathrin-coated pit (CCP) from the cell exterior. Here we used live-cell imaging of a pH-sensitive cargo to visualize the formation of clathrin-coated vesicles (CCVs) at single CCPs with a time resolution of seconds. We show that CCPs are highly dynamic and can produce multiple vesicles in succession. Using alternating evanescent field and epifluorescence illumination, we show that CCP invagination and scission are tightly coupled, with scission coinciding with maximal displacement of CCPs from the plasma membrane and with peak recruitment of cortactin-DsRed, a dynamin and F-actin binding protein. Finally, perturbing actin polymerization with latrunculin-B drastically reduces the efficiency of membrane scission and affects many aspects of CCP dynamics. We propose that CCP invagination, actin polymerization, and CCV formation are highly coordinated for efficient endocytosis.
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“Endocytosis typically requires force generation by active actin polymerization at the maturing endocytic pit52–57.”
From Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis (2023) · cites this paper as [56]
In the citing paper
“Together, these data strongly suggest that the observed heterogeneity in the dynamic behavior of CCPs (Merrifield et al., 2005; Rappoport et al., 2005; Yarar et al., 2005) could reflect differences in their content of cargo and adaptor molecules.”
From Cargo- and adaptor-specific mechanisms regulate clathrin-mediated endocytosis (2010) · cites this paper as Merrifield et al., 2005
In the citing paper
“This idea is further supported by the observation that nonterminal events are more frequent in HeLa cells (Ehrlich et al., 2004; Merrifield et al., 2005; Loerke et al., 2009), which express relatively high levels of Dab2 and cargo (Maurer and Cooper, 2006; unpublished data).”
From Cargo- and adaptor-specific mechanisms regulate clathrin-mediated endocytosis (2010) · cites this paper as Merrifield et al., 2005
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Cites
Curvature of clathrin-coated pits driven by epsin
Ford, M. G. J., Mills, I. G., Peter, B. J., Vallis, Y., Praefcke, G. J. K., Evans, P. R., & McMahon, H. T. (2002) · Nature · doi:10.1038/nature01020 · Cited by 1,019
Actin Assembly and Endocytosis: From Yeast to Mammals
Engqvist-Goldstein, Å. E. Y., & Drubin, D. G. (2003) · Annual Review of Cell and Developmental Biology · doi:10.1146/annurev.cellbio.19.111401.093127 · Cited by 572
Kirchhausen, T. (2000) · Annual Review of Biochemistry · doi:10.1146/annurev.biochem.69.1.699 · Cited by 560
Flagged first, then by citations
Research works printed but not linked: 44
- Blanpied, 2002, Dynamics and regulation of clathrin coats at specialized endocytic zones of dendrites and spines, Neuron, 36, 435, 10.1016/S0896-6273(02)00979-0 · DOI
- Bretscher, 1984, Endocytosis: relation to capping and cell locomotion, Science, 224, 681, 10.1126/science.6719108 · DOI
- Cao, 2003, Cortactin is a component of clathrin-coated pits and participates in receptor-mediated endocytosis, Mol. Cell. Biol., 23, 2162, 10.1128/MCB.23.6.2162-2170.2003 · DOI
- Daly, 2004, Cortactin signalling and dynamic actin networks, Biochem. J., 382, 13, 10.1042/BJ20040737 · DOI
- Ehrlich, 2004, Endocytosis by random initiation and stabilization of clathrin-coated pits, Cell, 118, 591, 10.1016/j.cell.2004.08.017 · DOI · Paper page
- Engqvist-Goldstein, 2001, The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro, J. Cell Biol., 154, 1209, 10.1083/jcb.200106089 · DOI
- Farsad, 2003, Mechanisms of membrane deformation, Curr. Opin. Cell Biol., 15, 372, 10.1016/S0955-0674(03)00073-5 · DOI
- Fujimoto, 2000, Actin assembly plays a variable, but not obligatory role in receptor-mediated endocytosis in mammalian cells, Traffic, 1, 161, 10.1034/j.1600-0854.2000.010208.x · DOI
- Gaidarov, 1999, Spatial control of coated-pit dynamics in living cells, Nat. Cell Biol., 1, 1, 10.1038/8971 · DOI
- Giardini, 2003, Compression forces generated by actin comet tails on lipid vesicles, Proc. Natl. Acad. Sci. USA, 100, 6493, 10.1073/pnas.1031670100 · DOI
- Heuser, 1980, Three-dimensional visualization of coated vesicle formation in fibroblasts, J. Cell Biol., 84, 560, 10.1083/jcb.84.3.560 · DOI
- Heuser, 1989, Effects of cytoplasmic acidification on clathrin lattice morphology, J. Cell Biol., 108, 401, 10.1083/jcb.108.2.401 · DOI
- Heuser, 1989, Hypertonic media inhibit receptor-mediated endocytosis by blocking clathrin-coated pit formation, J. Cell Biol., 108, 389, 10.1083/jcb.108.2.389 · DOI
- Higgins, 2002, Snap-shots of clathrin-mediated endocytosis, Trends Biochem. Sci., 27, 257, 10.1016/S0968-0004(02)02089-3 · DOI
- Iversen, 2003, Clathrin-coated pits with long, dynamin-wrapped necks upon expression of a clathrin antisense RNA, Proc. Natl. Acad. Sci. USA, 100, 5175, 10.1073/pnas.0534231100 · DOI
- Kaksonen, 2000, Association of cortactin with dynamic actin in lamellipodia and on endosomal vesicles, J. Cell Sci., 113, 4421, 10.1242/jcs.113.24.4421 · DOI
- Kaksonen, 2003, A pathway for association of receptors, adaptors, and actin during endocytic internalization, Cell, 115, 475, 10.1016/S0092-8674(03)00883-3 · DOI · Paper page
- Kessels, 2002, Syndapins integrate N-WASP in receptor-mediated endocytosis, EMBO J., 21, 6083, 10.1093/emboj/cdf604 · DOI
- Keyel, 2004, Endocytic adaptor molecules reveal an endosomal population of clathrin by total internal reflection fluorescence microscopy, J. Biol. Chem., 279, 13190, 10.1074/jbc.M312717200 · DOI
- McNiven, 2000, Regulated interaction between dynamin and the actin-binding protein cortactin modulate cell shape, J. Cell Biol., 151, 187, 10.1083/jcb.151.1.187 · DOI
- Merrifield, 2004, Seeing is believing: imaging actin dynamics at single sites of endocytosis, Trends Cell Biol., 14, 352, 10.1016/j.tcb.2004.05.008 · DOI
- Merrifield, 2002, Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits, Nat. Cell Biol., 4, 691, 10.1038/ncb837 · DOI · Paper page
- Merrifield, 2004, Neural Wiskott Aldrich Syndrome Protein (N-WASP) and the Arp2/3 complex are recruited to sites of clathrin-mediated endocytosis in cultured fibroblasts, Eur. J. Cell Biol., 83, 13, 10.1078/0171-9335-00356 · DOI
- Miesenbock, 1998, Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins, Nature, 394, 192, 10.1038/28190 · DOI
- Mise-Omata, 2003, Mammalian actin binding protein 1 is essential for endocytosis but not lamellipodia formation: functional analysis by RNA interference, Biochem. Biophys. Res. Commun., 301, 704, 10.1016/S0006-291X(02)02972-8 · DOI
- Mukherjee, 1997, Endocytosis, Physiol. Rev., 77, 759, 10.1152/physrev.1997.77.3.759 · DOI
- Perrais, 2004, Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells, J. Physiol., 560, 413, 10.1113/jphysiol.2004.064410 · DOI
- Praefcke, 2004, The dynamin superfamily: universal membrane tubulation and fission molecules?, Nat. Rev. Mol. Cell Biol., 5, 133, 10.1038/nrm1313 · DOI
- Qualmann, 2000, Molecular links between endocytosis and the actin cytoskeleton, J. Cell Biol., 150, F111, 10.1083/jcb.150.5.F111 · DOI
- Rappoport, 2003, The AP-2 complex is excluded from the dynamic population of plasma membrane-associated clathrin, J. Biol. Chem., 278, 47357, 10.1074/jbc.C300390200 · DOI
- Rappoport, 2004, Understanding living clathrin-coated pits, Traffic, 5, 327, 10.1111/j.1398-9219.2004.00187.x · DOI
- Sandvig, 1987, Acidification of the cytosol inhibits endocytosis from coated pits, J. Cell Biol., 105, 679, 10.1083/jcb.105.2.679 · DOI
- Sankaranarayanan, 2000, Real-time measurements of vesicle-SNARE recycling in synapses of the central nervous system, Nat. Cell Biol., 2, 197, 10.1038/35008615 · DOI
- Santini, 2002, A glimpse of coated vesicle creation? Well almost!, Nat. Cell Biol., 4, E230, 10.1038/ncb1002-e230 · DOI
- Schafer, 2002, Dynamin2 and cortactin regulate actin assembly and filament organization, Curr. Biol., 12, 1852, 10.1016/S0960-9822(02)01228-9 · DOI
- Schmid, 1991, Stage-specific assays for coated pit formation and coated vesicle budding in vitro, J. Cell Biol., 114, 869, 10.1083/jcb.114.5.869 · DOI
- Steyer, 2001, A real-time view of life within 100 nm of the plasma membrane, Nat. Rev. Mol. Cell Biol., 2, 268, 10.1038/35067069 · DOI
- Toomre, 2000, Fusion of constitutive membrane traffic with the cell surface observed by evanescent wave microscopy, J. Cell Biol., 149, 33, 10.1083/jcb.149.1.33 · DOI
- Upadhyaya, 2003, Probing polymerization forces by using actin-propelled lipid vesicles, Proc. Natl. Acad. Sci. USA, 100, 4521, 10.1073/pnas.0837027100 · DOI
- Weed, 2000, Cortactin localization to sites of actin assembly in lamellipodia requires interactions with F-actin and the Arp2/3 complex, J. Cell Biol., 151, 29, 10.1083/jcb.151.1.29 · DOI
- Willingham, 1983, Formation of receptosomes from plasma membrane coated pits during endocytosis: analysis by serial sections with improved membrane labeling and preservation techniques, Proc. Natl. Acad. Sci. USA, 80, 5617, 10.1073/pnas.80.18.5617 · DOI
- Willingham, 1981, Receptor-mediated endocytosis in cultured fibroblasts: cryptic coated pits and the formation of receptosomes, J. Histochem. Cytochem., 29, 1003, 10.1177/29.9.6169759 · DOI
- Yamashiro, 1984, Acidification of endocytic compartments and the intracellular pathways of ligands and receptors, J. Cell. Biochem., 26, 231, 10.1002/jcb.240260404 · DOI
- Yarar, 2004, A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis, Mol. Biol. Cell, 16, 964, 10.1091/mbc.E04-09-0774 · DOI · Paper page
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Items 401–433 of 433
Host cell entry of vesicular stomatitis virus
Jóhannsdóttir, H. K. (2008) · doi:10.3929/ethz-a-005692740 · Cited by 1
“The endocytic vesicle is then transported into the cytosol, possibly by the propelling force provided by polymerizing actin (177, 178).”
BackgroundRegulation of the Actin Cytoskeleton by Phospholipids
Skwarek‐Maruszewska, A., Hotulainen, P., & Lappalainen, P. (2006) · Advances in molecular and cell biology · doi:10.1016/s1569-2558(06)37010-5
Sposini, S., Rosendale, M., Claverie, L., Van, T. N. N., Jullié, D., & Perrais, D. (2020) · Nature Protocols · doi:10.1038/s41596-020-0371-z · Cited by 17
Self-citationInterview With Barbara Pearse and the Clathrin Family Tree
Brodsky, F. M., & Robinson, M. S. (2026) · Traffic · doi:10.1111/tra.70048
“Back in Cambridge that year, Scottie was working in Barbara's lab, separating and characterizing adaptor complexes [1, 47], while Barbara's advice helped Frances in California produce new monoclonal antibodies against the clathrin heavy chain that enabled identification of domains involved in…”
In situ mapping of late-stage biomechanical coordination during clathrin-mediated endocytosis
Wu, B., Luo, T., Zhu, Y., Zhang, X., Song, D., Lin, Y., Sun, Y., Huang, T.-X., Cheng, X.-D., & Fang, N. (2026) · Nature Communications · doi:10.1038/s41467-026-75876-y
“These steps are thought to be driven by the collective action of membrane curvature, protein scaffolding, and active force generation by molecular machines such as dynamin and the actin cytoskeleton 8 – 14 .”
Mechanistic Insight Into Clathrin‐Mediated Endocytosis in Plants
Zhu, T., Smet, I. D., & Damme, D. V. (2026) · Traffic · doi:10.1111/tra.70040 · Cited by 1
“Actin-related protein 2/3 (ARP2/3)- dependent actin polymerization generates the force to drive the CCV away from the PM in both yeast [69, 70] and mammalian cells [71, 72].”
Membrane lipid poly-unsaturation selectively affects dopamine D2 receptor endocytosis
Sposini, S., Baccouch, R., Lescuyer, M., Mali, A. A., Smedt, V. D., Heuninck, J., Gorše, A., Durroux, T., Trifilieff, P., Perrais, D., et al. (2026) · Nature Communications · doi:10.1038/s41467-026-73057-5
“Finally, we tested again with the ppH assay the speci fi city of PUFA enrichment on the endocytosis of the constitutive cargo for clathrin-mediated endocytosis, the transferrin receptor tagged with Lime (TfR-Lime) 48,49 .”
Self-citationMembrane lipid poly-unsaturation selectively affects dopamine D2 receptor internalizationPreprint
Sposini, S., Baccouch, R., Lescuyer, M., Mali, A., de Smedt-Peyrusse, V., Heuninck, J., Gorse, A., Durroux, T., Trifilieff, P., Perrais, D., et al. (2026) · bioRxiv · doi:10.1101/2023.12.14.571632
“Finally, we tested again with the ppH assay the specificity of PUFA enrichment on the endocytosis of the constitutive cargo for clathrin-mediated endocytosis, the transferrin receptor tagged with Lime (TfR-Lime) 46,47 .”
Self-citationSaenz, J., Yao, O., Aggarwal, M., Zhou, X., Barker, D. J., Dicicco-Bloom, E., & Pan, P.-Y. (2022) · bioRxiv · doi:10.1101/2022.10.12.511970
“The plasmid contains a CAT (combination of chicken beta-actin promoter with CMV enhancer) promotor and a KRas CAAX sequence at the C-terminus for improvement in plasma membrane targeting (Michaelson et al. 2005).”
BackgroundDynamin Action at the Final Stage of Clathrin-Mediated Endocytosis
Fang, N., Cheng, X.-D., Chen, K.-C., Dong, B., Yang, M., Filbrun, S., Myoung, Y., Huang, T.-X., Gu, Y., & Wang, G.-F. (2021) · doi:10.21203/rs.3.rs-134570/v1
“However, pinpointing the �ssion point in live cells has been a challenging problem 41,44 .”
BackgroundV-ATPase regulation of Hypoxia Inducible transcription Factors
Miles, A. L. (2018) · doi:10.17863/cam.30585
“65 HRE-mCherry (Generated by Ian Lobb, Nathan Lab) pDonor–MRP1-Clover-LoxP-Puro (gift from Dick van den Boomen) pIRE-YIC (Addgene 18784) Tfrn-pHl (gift from Christien Merrifield; Merrifield et al. 2005)”
MethodFranck, A. (2018) · theses.fr (ABES) · doi:10.70675/4ae507a1z8ee6z4b92z8a40z5838e4c5152f
“Using an elegant alternative pH system, Merrifield and colleagues have been able to detect if a clathrin-coated pit was open or closed depending on pH-sensitive fluorescence probes (Merrifield et al., 2005).”
BackgroundInfluentialPerrais, D., & Rosendale, M. (2017) · médecine/sciences · doi:10.1051/medsci/20173311009
Self-citationVisualisation and Perturbation of the Spatio-Temporal Dynamics of Endocytosis
Rosendale, M. (2015) · theses.fr (ABES) · doi:10.70675/c43e8a79z3005z46daz8eb6z6d7ab3a61888
“…selection and quantification of vesicles from data acquired with the ppH protocol are extensively described in the material and methods sections of (Merrifield et al., 2005; Perrais and Merrifield, 2005; Taylor et al., 2011) and each of the articles (published and in preparation) constituting the…”
BackgroundMethodResultInfluentialThe Function of V-ATPase in the Degradation of Gluconeogenic Enzymes in Yeast Vacuoles
Liu, J., & Chiang, H.-L. (2014) · doi:10.1201/b14984-4
Carroll, S., Stimpson, H. E. M., Weinberg, J., Toret, C. P., Sun, Y.-D., & Drubin, D. (2012) · Molecular Biology of the Cell · doi:10.1091/mboc.23.16.3275
Rôle de la clathrine dans la formation des lamellipodes
Gautier, J. J. (2011) · theses.fr (ABES) · doi:10.70675/1260cc02z60a9z4b51zaa64zf3641ef4752f
Visualization of Exocytosis and Endocytosis in Cultured Neurones Using TIRF Microscopy
Jullié, D., Moussa, W. A. B., & Perrais, D. (2011) · Microscopy and Microanalysis · doi:10.1017/s1431927611001097
“Using fast extracellular pH changes around the recorded cell [5], we show that these events are characterized by a short opening (median 2.”
BackgroundMethodSelf-citationアンフィファイジンとN-WASPのダイナミックな相互作用は，アクチン重合を制御する
Yamada, H., Padilla-Parra, S., Park, S. J., Itoh, T., Chaineau, M., Monaldi, I., Cremona, O., Benfenati, F., Camilli, P. D., Coppey-Moisan, M., et al. (2011) · doi:10.4044/joma.123.1
Perrais, D., Taylor, M. J., Jullié, D., & Merrifield, C. J. (2010) · doi:10.1017/s1431927610056308
“We have developed a protocol to detect the formation of clathrin coated vesicles (CCVs) from the plasma membrane of Swiss 3T3 cells (Merrifield et al., 2005) or hippocampal neurons.”
Self-citationLipid Domain Order and the Algebra of Morphology
Ursell, T., & Phillips, R. (2009) · doi:10.48550/arxiv.0905.1405
Bussell, K. (2005) · Nature reviews. Molecular cell biology · doi:10.1038/nrm1695
Endocytic recycling in Caenorhabditis elegans
Gleason, A. M. (2016) · doi:10.7282/t3s46v98
“A short-lived burst of actin polymerization at clathrin-coated pits has been reported to coincide with recruitment of the Arp2/3 complex (Merrifield et al., 2002; Merrifield et al., 2005).”
BackgroundNavaroli, D. M. (2012) · The Journal of the American Medical Association (JAMA) Network (American Medical Association) · doi:10.13028/5ajq-y522
The role of toca-1-N-wasp complex in filopodia formation and endocytosis
Bu, W. (2010) · DR-NTU (Nanyang Technological University) · doi:10.32657/10356/41562
“Thus during endocytosis, actin polymerization occurs at the site of clathrin-coated sites and stops after vesicle scission (Merrifield et al., 2005).”
BackgroundInfluentialBidirectional Crosstalk between Actin Dynamics and Endocytosis
Scita, G., & Fiore, P. P. D. (2010) · Elsevier eBooks · doi:10.1016/b978-0-12-374145-5.00305-3
Actin in Clathrin-Mediated Endocytosis
Kaksonen, M. (2010) · Springer eBooks · doi:10.1007/978-90-481-9301-1_4
Coupling Membrane Dynamics to Actin Polymerization
Suetsugu, S., & Takenawa, T. (2010) · Springer eBooks · doi:10.1007/978-90-481-9301-1_2
“The formation of clathrin-coated vesicles involves three steps (Kaksonen et al., 2005; Merrifield et al., 2005 ).”
Mechanisms of Endocytic Sorting: A Dissertation
Leonard, D. M. (2006) · The Journal of the American Medical Association (JAMA) Network (American Medical Association) · doi:10.13028/5c82-4606
Metadata
- Authors
- Merrifield, C. J., Perrais, D., & Zenisek, D.
- Affiliations
- MRC Laboratory of Molecular Biology (GB)
- Centre National de la Recherche Scientifique (FR)
- Yale University (US)
- Corresponding author
- Merrifield, Christien J.
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2005-05
- Volume
- 121
- Issue
- 4
- Pages
- 593-606
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2001532503
- Cited by
- 476
- References
- 0
- Citations by year
- 2026: 3 · 2025: 6 · 2024: 6 · 2023: 14 · 2022: 15 · 2021: 10 · 2020: 9 · 2019: 11 · 2018: 18 · 2017: 17 · 2016: 13 · 2015: 21 · 2014: 36 · 2013: 12 · 2012: 34
- FWCI
- 16.53
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Keywords
- clathrin-mediated endocytosis, membrane fission, clathrin-coated vesicles, cortactin, actin polymerization, latrunculin B, vesicle formation, dynamin, F-actin
- MeSH terms
- Actins, Animals, Cell Membrane, Coated Pits, Cell-Membrane, Endocytosis, Microfilament Proteins, Microscopy, Fluorescence, Polymers, Thiazoles, Microscopy, Video, Bridged Bicyclo Compounds, Heterocyclic, Clathrin-Coated Vesicles, Swiss 3T3 Cells, Cortactin, Mice, Thiazolidines
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