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.
Pubmed ID: 15992543 RIS Download
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