Characterization of a nonclathrin endocytic pathway

Membrane cargo and lipid requirements

Naava Naslavsky, Roberto Weigert, Julie G. Donaldson

Research output: Contribution to journalArticle

220 Citations (Scopus)

Abstract

Clathrin-independent endocytosis internalizes plasma membrane proteins that lack cytoplasmic sequences recognized by clathrin adaptor proteins. There is evidence for different clathrin-independent pathways but whether they share common features has not been systematically tested. Here, we examined whether CD59, an endogenous glycosylphosphatidyl inositol-anchored protein (GPI-AP), and major histocompatibility protein class I (MHCI), an endogenous, integral membrane protein, entered cells through a common mechanism and followed a similar itinerary. At early times of internalization, CD59 and MHCI were found in the same Arf6-associated endosomes before joining clathrin cargo proteins such as transferrin in common sorting endosomes. CD59 and MHCI, but not transferrin, also were observed in the Arf6-associated tubular recycling membranes. Endocytosis of CD59 and MHCI required free membrane cholesterol because it was inhibited by filipin binding to the cell surface. Expression of active Arf6 stimulated endocytosis of GPI-APs and MHCI to the same extent and led to their accumulation in Arf6 endosomes that labeled intensely with filipin. This blocked delivery of GPI-APs and MHCI to early sorting endosomes and to lysosomes for degradation. Endocytosis of transferrin was not affected by any of these treatments. These observations suggest common mechanisms for endocytosis without clathrin.

Original languageEnglish (US)
Pages (from-to)3542-3552
Number of pages11
JournalMolecular biology of the cell
Volume15
Issue number8
DOIs
StatePublished - Aug 1 2004

Fingerprint

Membrane Lipids
Histocompatibility
Endocytosis
Clathrin
Endosomes
Proteins
Transferrin
Filipin
Membrane Proteins
Vesicular Transport Adaptor Proteins
Membranes
Inositol
Lysosomes
Blood Proteins
Cholesterol
Cell Membrane

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Characterization of a nonclathrin endocytic pathway : Membrane cargo and lipid requirements. / Naslavsky, Naava; Weigert, Roberto; Donaldson, Julie G.

In: Molecular biology of the cell, Vol. 15, No. 8, 01.08.2004, p. 3542-3552.

Research output: Contribution to journalArticle

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