In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast

Thomas M. Newpher, Robin P. Smith, Vance Lemmon, Sandra K. Lemmon

Research output: Contribution to journalArticle

143 Scopus citations

Abstract

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 α 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.

Original languageEnglish (US)
Pages (from-to)87-98
Number of pages12
JournalDevelopmental Cell
Volume9
Issue number1
DOIs
StatePublished - Jul 1 2005

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Developmental Biology
  • Cell Biology

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