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.
Mesh Terms:
Actins, Adaptor Proteins, Signal Transducing, Adaptor Proteins, Vesicular Transport, Bicyclo Compounds, Heterocyclic, Carrier Proteins, Cell Membrane, Clathrin, Cytoskeletal Proteins, Cytoskeleton, Endocytosis, Endosomes, Microfilament Proteins, Microscopy, Fluorescence, Monomeric Clathrin Assembly Proteins, Mutation, Receptors, Mating Factor, Receptors, Peptide, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Thiazoles, Thiazolidines, Transcription Factors, Vesicular Transport Proteins
Dev. Cell
Date: Jul. 01, 2005
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