A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis

Defne Yarar, Clare M. Waterman-Storer, Sandra L. Schmid

Research output: Contribution to journalArticle

309 Citations (Scopus)

Abstract

Clathrin-mediated endocytosis in mammalian cells is critical for a variety of cellular processes including nutrient uptake and cell surface receptor down-regulation. Despite the findings that numerous endocytic accessory proteins directly or indirectly regulate actin dynamics and that actin assembly is spatially and temporally coordinated with endocytosis, direct functional evidence for a role of actin during clathrin-coated vesicle formation is lacking. Here, we take parallel biochemical and microscopic approaches to address the contribution of actin polymerization/depolymerization dynamics to clathrin-mediated endocytosis. When measured using live-cell fluorescence microscopy, disruption of the F-actin assembly and disassembly cycle with latrunculin A or jasplakinolide results in near complete cessation of all aspects of clathrin-coated structure (CCS) dynamics. Stage-specific biochemical assays and quantitative fluorescence and electron microscopic analyses establish that F-actin dynamics are required for multiple distinct stages of clathrin-coated vesicle formation, including coated pit formation, constriction, and internalization. In addition, F-actin dynamics are required for observed diverse CCS behaviors, including splitting of CCSs from larger CCSs, merging of CCSs, and lateral mobility on the cell surface. Our results demonstrate a key role for actin during clathrin-mediated endocytosis in mammalian cells.

Original languageEnglish (US)
Pages (from-to)964-975
Number of pages12
JournalMolecular Biology of the Cell
Volume16
Issue number2
DOIs
StatePublished - Feb 2005

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Clathrin
Endocytosis
Actin Cytoskeleton
Actins
Clathrin-Coated Vesicles
jasplakinolide
Cell Surface Receptors
Fluorescence Microscopy
Constriction
Polymerization
Down-Regulation
Fluorescence
Electrons

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

A dynamic actin cytoskeleton functions at multiple stages of clathrin-mediated endocytosis. / Yarar, Defne; Waterman-Storer, Clare M.; Schmid, Sandra L.

In: Molecular Biology of the Cell, Vol. 16, No. 2, 02.2005, p. 964-975.

Research output: Contribution to journalArticle

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