Endocytic trafficking routes of wild type and ΔF508 cystic fibrosis transmembrane conductance regulator

Martina Gentzsch, Xiu-Bao D Chang, Liying Cui, Yufeng Wu, Victor V. Ozols, Amit Choudhury, Richard E. Pagano, John R. Riordan

Research output: Contribution to journalArticle

165 Citations (Scopus)

Abstract

Intracellular trafficking of cystic fibrosis transmembrane conductance regulator (CFTR) is a focus of attention because it is defective in most patients with cystic fibrosis. ΔF508 CFTR, which does not mature conformationally, normally does not exit the endoplasmic reticulum, but if induced to do so at reduced temperature is short-lived at the surface. We used external epitope-tagged constructs to elucidate the itinerary and kinetics of wild type and ΔF508 CFTR in the endocytic pathway and visualized movement of CFTR from the surface to intracellular compartments. Modulation of different endocytic steps with low temperature (16°C) block, protease inhibitors, and overexpression of wild type and mutant Rab GTPases revealed that surface CFTR enters several different routes, including a Rab5-dependent initial step to early endosomes, then either Rab11-dependent recycling back to the surface or Rab7-regulated movement to late endosomes or alternatively Rab9-mediated transit to the trans-Golgi network. Without any of these modulations ΔF508 CFTR rapidly disappears from and does not return to the cell surface, confirming that its altered structure is detected in the distal as well as proximal secretory pathway. Importantly, however, the mutant protein can be rescued at the plasma membrane by Rab11 overexpression, proteasome inhibitors, or inhibition of Rab5-dependent endocytosis.

Original languageEnglish (US)
Pages (from-to)2684-2696
Number of pages13
JournalMolecular Biology of the Cell
Volume15
Issue number6
DOIs
StatePublished - Jun 2004

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Cystic Fibrosis Transmembrane Conductance Regulator
Endosomes
rab GTP-Binding Proteins
trans-Golgi Network
Proteasome Inhibitors
Temperature
Secretory Pathway
Recycling
Mutant Proteins
Endocytosis
Protease Inhibitors
Cystic Fibrosis
Endoplasmic Reticulum
Epitopes
Cell Membrane

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Gentzsch, M., Chang, X-B. D., Cui, L., Wu, Y., Ozols, V. V., Choudhury, A., ... Riordan, J. R. (2004). Endocytic trafficking routes of wild type and ΔF508 cystic fibrosis transmembrane conductance regulator. Molecular Biology of the Cell, 15(6), 2684-2696. https://doi.org/10.1091/mbc.E04-03-0176

Endocytic trafficking routes of wild type and ΔF508 cystic fibrosis transmembrane conductance regulator. / Gentzsch, Martina; Chang, Xiu-Bao D; Cui, Liying; Wu, Yufeng; Ozols, Victor V.; Choudhury, Amit; Pagano, Richard E.; Riordan, John R.

In: Molecular Biology of the Cell, Vol. 15, No. 6, 06.2004, p. 2684-2696.

Research output: Contribution to journalArticle

Gentzsch, M, Chang, X-BD, Cui, L, Wu, Y, Ozols, VV, Choudhury, A, Pagano, RE & Riordan, JR 2004, 'Endocytic trafficking routes of wild type and ΔF508 cystic fibrosis transmembrane conductance regulator', Molecular Biology of the Cell, vol. 15, no. 6, pp. 2684-2696. https://doi.org/10.1091/mbc.E04-03-0176
Gentzsch, Martina ; Chang, Xiu-Bao D ; Cui, Liying ; Wu, Yufeng ; Ozols, Victor V. ; Choudhury, Amit ; Pagano, Richard E. ; Riordan, John R. / Endocytic trafficking routes of wild type and ΔF508 cystic fibrosis transmembrane conductance regulator. In: Molecular Biology of the Cell. 2004 ; Vol. 15, No. 6. pp. 2684-2696.
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