A PDZ-binding motif is essential but not sufficient to localize the C terminus of CFTR to the apical membrane

Michal I. Milewski, John E. Mickle, John K. Forrest, Diane M. Stafford, Bryan D. Moyer, Jie Cheng, William B Guggino, Bruce A. Stanton, Garry R Cutting

Research output: Contribution to journalArticle

Abstract

Localization of ion channels and transporters to the correct membrane of polarized epithelia is important for vectorial ion movement. Prior studies have shown that the cytoplasmic carboxyl terminus of the cystic fibrosis transmembrane conductance regulator (CFTR) is involved in the apical localization of this protein. Here we show that the C-terminal tail alone, or when fused to the green fluorescent protein (GFP), can localize to the apical plasma membrane, despite the absence of transmembrane domains. Co-expression of the C terminus with full-length CFTR results in redistribution of CFTR from apical to basolateral membranes, indicating that both proteins interact with the same target at the apical membrane. Amino acid substitution and deletion analysis confirms the importance of a PDZ-binding motif D-T-R-L> for apical localization. However, two other C-terminal regions, encompassing amino acids 1370-1394 and 1404-1425 of human CFTR, are also required for localizing to the apical plasma membrane. Based on these results, we propose a model of polarized distribution of CFTR, which includes a mechanism of selective retention of this protein in the apical plasma membrane and stresses the requirement for other C-terminal sequences in addition to a PDZ-binding motif.

Original languageEnglish (US)
Pages (from-to)719-726
Number of pages8
JournalJournal of Cell Science
Volume114
Issue number4
StatePublished - 2001

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Membranes
Cell Membrane
Proteins
Amino Acid Substitution
Green Fluorescent Proteins
Ion Channels
Tail
Epithelium
Ions
Amino Acids

Keywords

  • Apical polarization
  • CFTR
  • Cystic fibrosis
  • Localization signal
  • PDZ domain

ASJC Scopus subject areas

  • Cell Biology

Cite this

A PDZ-binding motif is essential but not sufficient to localize the C terminus of CFTR to the apical membrane. / Milewski, Michal I.; Mickle, John E.; Forrest, John K.; Stafford, Diane M.; Moyer, Bryan D.; Cheng, Jie; Guggino, William B; Stanton, Bruce A.; Cutting, Garry R.

In: Journal of Cell Science, Vol. 114, No. 4, 2001, p. 719-726.

Research output: Contribution to journalArticle

Milewski, MI, Mickle, JE, Forrest, JK, Stafford, DM, Moyer, BD, Cheng, J, Guggino, WB, Stanton, BA & Cutting, GR 2001, 'A PDZ-binding motif is essential but not sufficient to localize the C terminus of CFTR to the apical membrane', Journal of Cell Science, vol. 114, no. 4, pp. 719-726.
Milewski, Michal I. ; Mickle, John E. ; Forrest, John K. ; Stafford, Diane M. ; Moyer, Bryan D. ; Cheng, Jie ; Guggino, William B ; Stanton, Bruce A. ; Cutting, Garry R. / A PDZ-binding motif is essential but not sufficient to localize the C terminus of CFTR to the apical membrane. In: Journal of Cell Science. 2001 ; Vol. 114, No. 4. pp. 719-726.
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AU - Milewski, Michal I.

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AU - Forrest, John K.

AU - Stafford, Diane M.

AU - Moyer, Bryan D.

AU - Cheng, Jie

AU - Guggino, William B

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AU - Cutting, Garry R

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AB - Localization of ion channels and transporters to the correct membrane of polarized epithelia is important for vectorial ion movement. Prior studies have shown that the cytoplasmic carboxyl terminus of the cystic fibrosis transmembrane conductance regulator (CFTR) is involved in the apical localization of this protein. Here we show that the C-terminal tail alone, or when fused to the green fluorescent protein (GFP), can localize to the apical plasma membrane, despite the absence of transmembrane domains. Co-expression of the C terminus with full-length CFTR results in redistribution of CFTR from apical to basolateral membranes, indicating that both proteins interact with the same target at the apical membrane. Amino acid substitution and deletion analysis confirms the importance of a PDZ-binding motif D-T-R-L> for apical localization. However, two other C-terminal regions, encompassing amino acids 1370-1394 and 1404-1425 of human CFTR, are also required for localizing to the apical plasma membrane. Based on these results, we propose a model of polarized distribution of CFTR, which includes a mechanism of selective retention of this protein in the apical plasma membrane and stresses the requirement for other C-terminal sequences in addition to a PDZ-binding motif.

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