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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

Authors

  • Newpher TM
  • Smith RP
  • Lemmon V
  • Lemmon SK

Journal

Developmental cell

Publication Data

July 4, 2005

Associated Grants

  • Agency: NEI NIH HHS, Id: R01 EY005285
  • Agency: NEI NIH HHS, Id: R01 EY05285
  • Agency: NIGMS NIH HHS, Id: R01 GM55796
  • Agency: NIGMS NIH HHS, Id: T32 GM08056-20

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