Molecular basis of oligoubiquitin-dependent internalization of membrane proteins in mammalian cells

Herve Barriere, Csilla Nemes, Delphine Lechardeur, Mina Khan-Mohammad, Klaus Frueh, Gergely L. Lukacs

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Ubiquitination induced down-regulation of cell surface proteins by internalization and lysosomal targeting plays a fundamental role in cell physiology and pathogenesis of diseases. The molecular basis of a single ubiquitin (Ub) as an autonomous endocytic signal, the widely accepted mechanism, however, remains elusive in higher eukaryotes. Using Ub containing reporter proteins without signalling abilities, we present evidence that only multiple Ub moieties, linked either covalently or assembled as oligomers with an intact interface for recognition by Ub-interacting motifs (UIMs), are recognized by the endocytic machinery in vivo and associate with a subset of Ub-binding clathrin adaptors in vitro. Genetic and pharmacological approaches show that internalization of plasma membrane proteins harbouring multiple Ub moieties is clathrin-dependent, but caveolinindependent. Functional assays demonstrate the cargodependent involvement of eps15/15R and epsin, UIM containing clathrin adaptors, in the endocytosis of model proteins, CD4 and the activated β2-adrenergic receptor complex, containing polymeric or oligomeric Ub. These results provide a paradigm for the clathrinmediated uptake of ubiquitinated membrane proteins in mammalian cells, requiring the assembly of multiple UIM-Ub interactions to overcome the low affinity binding of mono-Ub to UIM.

Original languageEnglish (US)
Pages (from-to)282-297
Number of pages16
JournalTraffic
Volume7
Issue number3
DOIs
StatePublished - Mar 2006

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Ubiquitin
Membrane Proteins
Cells
Vesicular Transport Adaptor Proteins
Ubiquitinated Proteins
Cell Physiological Phenomena
Clathrin
Ubiquitination
Physiology
Cell membranes
Endocytosis
Eukaryota
Oligomers
Adrenergic Receptors
Machinery
Blood Proteins
Assays
Proteins
Down-Regulation
Cell Membrane

Keywords

  • β-adrenergic receptor
  • Caveolin
  • CD4
  • Clathrin
  • Endocytosis
  • Eps15
  • Epsin
  • Ubiquitin
  • UIM

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Structural Biology
  • Molecular Biology
  • Genetics

Cite this

Molecular basis of oligoubiquitin-dependent internalization of membrane proteins in mammalian cells. / Barriere, Herve; Nemes, Csilla; Lechardeur, Delphine; Khan-Mohammad, Mina; Frueh, Klaus; Lukacs, Gergely L.

In: Traffic, Vol. 7, No. 3, 03.2006, p. 282-297.

Research output: Contribution to journalArticle

Barriere, Herve ; Nemes, Csilla ; Lechardeur, Delphine ; Khan-Mohammad, Mina ; Frueh, Klaus ; Lukacs, Gergely L. / Molecular basis of oligoubiquitin-dependent internalization of membrane proteins in mammalian cells. In: Traffic. 2006 ; Vol. 7, No. 3. pp. 282-297.
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