In Vivo Dynamics of Clathrin and Its Adaptor-Dependent Recruitment to the Actin-Based Endocytic Machinery in Yeast
In Vivo Dynamics of Clathrin and Its Adaptor-Dependent Recruitment to the Actin-Based Endocytic Machinery in Yeast
Clathrin-mediated transport is a major pathway for endocytosis. However, in yeast, where cortical actin patches are essential for endocytosis, plasma membrane-associated clathrin has never been observed. Using live cell imaging, we demonstrate cortical clathrin in association with the actin-based endocytic machinery in yeast. Fluorescently tagged clathrin is found in highly mobile internal trans-Golgi/endosomal structures and in smaller cortical patches. Total internal reflection fluorescence microscopy showed that cortical patches are likely endocytic sites, as clathrin is recruited prior to a burst of intensity of the actin patch/endocytic marker, Abp1. Clathrin also accumulates at the cortex with internalizing alpha factor receptor, Ste2p. Cortical clathrin localizes with epsins Ent1/2p and AP180s, and its recruitment to the surface is dependent upon these adaptors. In contrast, Sla2p, End3p, Pan1p, and a dynamic actin cytoskeleton are not required for clathrin assembly or exchange but are required for the mobility, maturation, and/or turnover of clathrin-containing endocytic structures.
- Miami University United States
- Duke University United States
- Case Western Reserve University United States
- CASE WESTERN RESERVE UNIVERSITY
- Miami Project to Cure Paralysis, Miami, FL, USA United States
570, Saccharomyces cerevisiae Proteins, Receptors, Peptide, Vesicular Transport Proteins, Endosomes, Saccharomyces cerevisiae, Fluorescence, Bridged Bicyclo Compounds, Receptors, Cytoskeleton, Adaptor Proteins, Signal Transducing, Microscopy, Heterocyclic, Cell Membrane, Microfilament Proteins, Signal Transducing, Adaptor Proteins, Bridged Bicyclo Compounds, Heterocyclic, Actins, Clathrin, Endocytosis, Vesicular Transport, Adaptor Proteins, Vesicular Transport, Cytoskeletal Proteins, Thiazoles, Microscopy, Fluorescence, Monomeric Clathrin Assembly Proteins, Peptide, Mutation, Receptors, Mating Factor, Thiazolidines, Mating Factor, Carrier Proteins, Transcription Factors, Developmental Biology
570, Saccharomyces cerevisiae Proteins, Receptors, Peptide, Vesicular Transport Proteins, Endosomes, Saccharomyces cerevisiae, Fluorescence, Bridged Bicyclo Compounds, Receptors, Cytoskeleton, Adaptor Proteins, Signal Transducing, Microscopy, Heterocyclic, Cell Membrane, Microfilament Proteins, Signal Transducing, Adaptor Proteins, Bridged Bicyclo Compounds, Heterocyclic, Actins, Clathrin, Endocytosis, Vesicular Transport, Adaptor Proteins, Vesicular Transport, Cytoskeletal Proteins, Thiazoles, Microscopy, Fluorescence, Monomeric Clathrin Assembly Proteins, Peptide, Mutation, Receptors, Mating Factor, Thiazolidines, Mating Factor, Carrier Proteins, Transcription Factors, Developmental Biology
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