Clathrin-mediated endocytosis is an essential process that forms vesicles from the plasma membrane. Although most of the protein components of the endocytic protein machinery have been thoroughly characterized, their organization at the endocytic site is poorly understood. We developed a fluorescence microscopy method to track the average positions of yeast endocytic proteins in relation to each other with a time precision below 1 s and with a spatial precision of ~10 nm. With these data, integrated with shapes of endocytic membrane intermediates and with superresolution imaging, we could visualize the dynamic architecture of the endocytic machinery. We showed how different coat proteins are distributed within the coat structure and how the assembly dynamics of N-BAR proteins relate to membrane shape changes. Moreover, we found that the region of actin polymerization is located at the base of the endocytic invagination, with the growing ends of filaments pointing toward the plasma membrane.
The END3 gene encodes a protein that is required for the internalization step of endocytosis and for actin cytoskeleton organization in yeast
Synthetic-lethal interactions identify two novel genes, SLA1 and SLA2, that control membrane cytoskeleton assembly in Saccharomyces cerevisiae
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.
A role for myosin-I in actin assembly through interactions with Vrp1p, Bee1p, and the Arp2/3 complex
The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro
An interaction between Sla1p and Sla2p plays a role in regulating actin dynamics and endocytosis in budding yeast
Interaction of Sla2p's ANTH domain with PtdIns(4,5)P2 is important for actin-dependent endocytic internalization
In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast
Dissection of Arp2/3 complex actin nucleation mechanism and distinct roles for its nucleation-promoting factors in Saccharomyces cerevisiae
Endocytic internalization in budding yeast requires coordinated actin nucleation and myosin motor activity
Differential evanescence nanometry: live-cell fluorescence measurements with 10-nm axial resolution on the plasma membrane
The direction of actin polymerization for vesicle fission suggested from membranes tubulated by the EFC/F-BAR domain protein FBP17
Multiple modes of endophilin-mediated conversion of lipid vesicles into coated tubes: implications for synaptic endocytosis.
Mathematical modeling of endocytic actin patch kinetics in fission yeast: disassembly requires release of actin filament fragments
Quantitative analysis of the mechanism of endocytic actin patch assembly and disassembly in fission yeast
Clathrin light chain directs endocytosis by influencing the binding of the yeast Hip1R homologue, Sla2, to F-actin
Distinct roles for F-BAR proteins Cdc15p and Bzz1p in actin polymerization at sites of endocytosis in fission yeast
Recruitment of endophilin to clathrin-coated pit necks is required for efficient vesicle uncoating after fission
Membrane fission is promoted by insertion of amphipathic helices and is restricted by crescent BAR domains
Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis
Actin filament severing by cofilin dismantles actin patches and produces mother filaments for new patches
Local and global analysis of endocytic patch dynamics in fission yeast using a new "temporal superresolution" realignment method
Avoiding artefacts when counting polymerized actin in live cells with LifeAct fused to fluorescent proteins
High-speed superresolution imaging of the proteins in fission yeast clathrin-mediated endocytic actin patches
Control of actin polymerization via the coincidence of phosphoinositides and high membrane curvature
Crowd-Sourcing of Membrane Fission: How crowding of non-specialized membrane-bound proteins contributes to cellular membrane fission
Higher-order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle
Single-molecule turnover dynamics of actin and membrane coat proteins in clathrin-mediated endocytosis
A primer on resolving the nanoscale structure of the plasma membrane with light and electron microscopy
Three members of the yeast N-BAR proteins family form heterogeneous lattices in vivo and interact differentially with two RabGAP proteins.
A low-cost, large field-of-view scanning ion conductance microscope for studying nanoparticle-cell membrane interactions
The S. pombe adaptor protein Bbc1 regulates localization of Wsp1 and Vrp1 during endocytic actin patch assembly
Direct comparison of clathrin-mediated endocytosis in budding and fission yeast reveals conserved and evolvable features
Neurodevelopmental disease mechanisms, primary cilia, and endosomes converge on the BLOC-1 and BORC complexes
Principles of self-organization and load adaptation by the actin cytoskeleton during clathrin-mediated endocytosis.
Quantitative mapping of fluorescently tagged cellular proteins using FCS-calibrated four-dimensional imaging
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