| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
F508 CFTR in CFBE41o– airway epithelial monolayers
1 Gregory Fleming James Cystic Fibrosis Research Center
2 Department of Cell Biology
5 Department of Pediatrics
6 Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA
3 Tranzyme, Inc., Birmingham, AL, USA
4 Southern Research Institute, Birmingham, AL, USA
The cystic fibrosis transmembrane conductance regulator (CFTR) is a cyclic AMP-regulated chloride channel. Mutations in the CFTR gene result in cystic fibrosis (CF). The most common mutation, 
F508, results in endoplasmic reticulum-associated degradation (ERAD) of CFTR. F508 CFTR has been described as a temperature-sensitive mutation that can be rescued following growth at 27°C. In order to study the processing and function of wild-type and rescued F508 CFTR at the cell surface under non-polarized and polarized conditions, we developed stable cell lines expressing F508 or wild-type CFTR. CFBE41o– is a human airway epithelial cell line capable of forming high resistance, polarized monolayers when cultured on permeable supports, while HeLa cells are normally grown under non-polarizing conditions. Immunoprecipitation, cell surface biotinylation, immunofluorescence, and functional assays confirmed the presence of F508 CFTR at the cell surface in both cell lines after incubating the cells for 48 h at 27°C. However, stimulators of wild-type CFTR such as forskolin, ß2-adrenergic or A2B-adenosine receptor agonists failed to activate rescued F508 CFTR in CFBE41o– monolayers. Rescued F508 CFTR could be stimulated with genistein independent of pretreatment with cAMP signalling agonists. Interestingly, rescued F508 CFTR in HeLa cells could be efficiently stimulated with either forskolin or genistein to promote Cl– transport. These results indicate that F508 CFTR, when rescued in CFBE41o– human airway epithelial cells, is poorly responsive to signalling pathways known to regulate wild-type CFTR. Furthermore, the differences in rescue and activation of F508 CFTR in the two cell lines suggest that cell-type specific differences in F508 CFTR processing are likely to complicate efforts to identify potentiators and/or correctors of the F508 defect.
(Received 17 August 2005;
accepted after revision 4 October 2005;
first published online 6 October 2005)
Corresponding author J. P. Clancy: Department of Pediatrics, University of Alabama at Birmingham, 620 ACC, 1600 7th Ave South, Birmingham, AL 35233, USA. Email: jpclancy{at}peds.uab.edu
This article has been cited by other articles:
|
|
 |
|
  R. Bartoszewski, A. Rab, A. Jurkuvenaite, M. Mazur, J. Wakefield, J. F. Collawn, and Z. Bebok Activation of the Unfolded Protein Response by {Delta}F508 CFTR Am. J. Respir. Cell Mol. Biol., October 1, 2008; 39(4): 448 - 457. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Cebotaru, N. Vij, I. Ciobanu, J. Wright, T. Flotte, and W. B. Guggino Cystic Fibrosis Transmembrane Regulator Missing the First Four Transmembrane Segments Increases Wild Type and {Delta}F508 Processing J. Biol. Chem., August 8, 2008; 283(32): 21926 - 21933. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Bartoszewski, A. Rab, G. Twitty, L. Stevenson, J. Fortenberry, A. Piotrowski, J. P. Dumanski, and Z. Bebok The Mechanism of Cystic Fibrosis Transmembrane Conductance Regulator Transcriptional Repression during the Unfolded Protein Response J. Biol. Chem., May 2, 2008; 283(18): 12154 - 12165. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Robert, G. W. Carlile, C. Pavel, N. Liu, S. M. Anjos, J. Liao, Y. Luo, D. Zhang, D. Y. Thomas, and J. W. Hanrahan Structural Analog of Sildenafil Identified as a Novel Corrector of the F508del-CFTR Trafficking Defect Mol. Pharmacol., February 1, 2008; 73(2): 478 - 489. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Rowe, K. Varga, A. Rab, Z. Bebok, K. Byram, Y. Li, E. J. Sorscher, and J. P. Clancy Restoration of W1282X CFTR Activity by Enhanced Expression Am. J. Respir. Cell Mol. Biol., September 1, 2007; 37(3): 347 - 356. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. A. Borthwick, J. Mcgaw, G. Conner, C. J. Taylor, V. Gerke, A. Mehta, L. Robson, and R. Muimo The Formation of the cAMP/Protein Kinase A-dependent Annexin 2 S100A10 Complex with Cystic Fibrosis Conductance Regulator Protein (CFTR) Regulates CFTR Channel Function Mol. Biol. Cell, September 1, 2007; 18(9): 3388 - 3397. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Swiatecka-Urban, L. Talebian, E. Kanno, S. Moreau-Marquis, B. Coutermarsh, K. Hansen, K. H. Karlson, R. Barnaby, R. E. Cheney, G. M. Langford, et al. Myosin Vb Is Required for Trafficking of the Cystic Fibrosis Transmembrane Conductance Regulator in Rab11a-specific Apical Recycling Endosomes in Polarized Human Airway Epithelial Cells J. Biol. Chem., August 10, 2007; 282(32): 23725 - 23736. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. P. MacEachran, S. Ye, J. M. Bomberger, D. A. Hogan, A. Swiatecka-Urban, B. A. Stanton, and G. A. O'Toole The Pseudomonas aeruginosa Secreted Protein PA2934 Decreases Apical Membrane Expression of the Cystic Fibrosis Transmembrane Conductance Regulator Infect. Immun., August 1, 2007; 75(8): 3902 - 3912. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Wolde, A. Fellows, J. Cheng, A. Kivenson, B. Coutermarsh, L. Talebian, K. Karlson, A. Piserchio, D. F. Mierke, B. A. Stanton, et al. Targeting CAL as a Negative Regulator of {Delta}F508-CFTR Cell-Surface Expression: AN RNA INTERFERENCE AND STRUCTURE-BASED MUTAGENETIC APPROACH J. Biol. Chem., March 16, 2007; 282(11): 8099 - 8109. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Rab, R. Bartoszewski, A. Jurkuvenaite, J. Wakefield, J. F. Collawn, and Z. Bebok Endoplasmic reticulum stress and the unfolded protein response regulate genomic cystic fibrosis transmembrane conductance regulator expression Am J Physiol Cell Physiol, February 1, 2007; 292(2): C756 - C766. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Wang, M. C. Bartlett, T. W. Loo, and D. M. Clarke Specific Rescue of Cystic Fibrosis Transmembrane Conductance Regulator Processing Mutants Using Pharmacological Chaperones Mol. Pharmacol., July 1, 2006; 70(1): 297 - 302. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Li, W. Wang, W. Parker, and J. P. Clancy Adenosine Regulation of Cystic Fibrosis Transmembrane Conductance Regulator through Prostenoids in Airway Epithelia Am. J. Respir. Cell Mol. Biol., May 1, 2006; 34(5): 600 - 608. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Swiatecka-Urban, S. Moreau-Marquis, D. P. MacEachran, J. P. Connolly, C. R. Stanton, J. R. Su, R. Barnaby, G. A. O'Toole, and B. A. Stanton Pseudomonas aeruginosa inhibits endocytic recycling of CFTR in polarized human airway epithelial cells Am J Physiol Cell Physiol, March 1, 2006; 290(3): C862 - C872. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. R. Hallows, A. C. Fitch, C. A. Richardson, P. R. Reynolds, J. P. Clancy, P. C. Dagher, L. A. Witters, J. K. Kolls, and J. M. Pilewski Up-regulation of AMP-activated Kinase by Dysfunctional Cystic Fibrosis Transmembrane Conductance Regulator in Cystic Fibrosis Airway Epithelial Cells Mitigates Excessive Inflammation J. Biol. Chem., February 17, 2006; 281(7): 4231 - 4241. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
