Phosphatidylinositol-(4,5)-bisphosphate regulates clathrin-coated pit initiation, stabilization, and size

Costin N. Antonescu, François Aguet, Gaudenz Danuser, Sandra L. Schmid

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Clathrin-mediated endocytosis (CME) is the major mechanism for internalization in mammalian cells. CME initiates by recruitment of adaptors and clathrin to form clathrin-coated pits (CCPs). Nearly half of nascent CCPs abort, whereas others are stabilized by unknown mechanisms and undergo further maturation before pinching off to form clathrin-coated vesicles (CCVs). Phosphatidylinositol-(4,5)-bisphosphate (PIP 2), the main lipid binding partner of endocytic proteins, is required for CCP assembly, but little is currently known about its contribution(s) to later events in CCV formation. Using small interfering RNA (siRNA) knockdown and overexpression, we have analyzed the effects of manipulating PIP 2 synthesis and turnover on CME by quantitative total internal reflection fluorescence microscopy and computational analysis. Phosphatidylinositol-4-phosphate-5-kinase cannot be detected within CCPs but functions in initiation and controls the rate and extent of CCP growth. In contrast, the 5′-inositol phosphatase synaptojanin 1 localizes to CCPs and controls early stabilization and maturation efficiency. Together these results suggest that the balance of PIP 2 synthesis in the bulk plasma membrane and its local turnover within CCPs control multiple stages of CCV formation.

Original languageEnglish (US)
Pages (from-to)2588-2600
Number of pages13
JournalMolecular Biology of the Cell
Volume22
Issue number14
DOIs
StatePublished - Jul 15 2011

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Clathrin
Phosphatidylinositols
Clathrin-Coated Vesicles
Endocytosis
Vesicular Transport Adaptor Proteins
Fluorescence Microscopy
Small Interfering RNA
Cell Membrane
Lipids

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Phosphatidylinositol-(4,5)-bisphosphate regulates clathrin-coated pit initiation, stabilization, and size. / Antonescu, Costin N.; Aguet, François; Danuser, Gaudenz; Schmid, Sandra L.

In: Molecular Biology of the Cell, Vol. 22, No. 14, 15.07.2011, p. 2588-2600.

Research output: Contribution to journalArticle

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