Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR

L S Ostedgaard; B Zeiher; M J Welsh

doi:10.1242/jcs.112.13.2091

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Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR

Journal article

Peer reviewed

Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR

L S Ostedgaard, B Zeiher and M J Welsh

Journal of cell science, Vol.112 (13), pp.2091-2098

07/1999

DOI: 10.1242/jcs.112.13.2091

PMID: 10362539

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Abstract

Cystic fibrosis transmembrane conductance regulator (CFTR) containing the deltaF508 mutation is retained in the endoplasmic reticulum (ER). This defect can be partially overcome by a reduction in temperature which allows some of the deltaF508 protein to exit the ER and move to the cell surface. Earlier studies showed that the CF-associated mutants, P574H and A455E, were also misprocessed. In this study, we found that processing of P574H and A455E was also temperature-sensitive; at 26 degrees C, some of the protein matured. In contrast to other CFTR mutants, P574H accumulated in punctate cytoplasmic bodies that colocalized with endoplasmic reticulum (ER) markers. At 26 degrees C, these bodies were no longer present. P574H showed a prolonged association with Hsp70 and also colocalized with Hsp70. We used brefeldin A (BFA) to determine which processing step(s) was altered by reduced temperature. Unlike wild-type CFTR, which was converted into an intermediate that was stable in the presence of BFA at 37 degrees C, deltaF508 and P574H produced the intermediate only when the temperature was reduced to 26 degrees C. Furthermore the wild-type intermediate was not associated with Hsp70. These data suggest that formation of the stable intermediate is a key temperature-sensitive step and appears to be coincident with release of the wild-type protein from Hsp70.

Sequence Deletion

Temperature

Mutagenesis, Site-Directed

Cystic Fibrosis - metabolism

Golgi Apparatus - drug effects

Humans

Endoplasmic Reticulum - metabolism

Brefeldin A - pharmacology

Cystic Fibrosis Transmembrane Conductance Regulator - metabolism

HSP70 Heat-Shock Proteins - metabolism

Point Mutation

Animals

Protein Processing, Post-Translational - drug effects

Cystic Fibrosis - genetics

Endoplasmic Reticulum - drug effects

Cystic Fibrosis Transmembrane Conductance Regulator - genetics

Golgi Apparatus - metabolism

Protein Synthesis Inhibitors - pharmacology

COS Cells

Details

Title: Subtitle: Processing of CFTR bearing the P574H mutation differs from wild-type and deltaF508-CFTR
Creators: L S Ostedgaard - Howard Hughes Medical Institute, Departments of Internal Medicine and Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA
B Zeiher
M J Welsh
Resource Type: Journal article
Publication Details: Journal of cell science, Vol.112 (13), pp.2091-2098
Publisher: England
DOI: 10.1242/jcs.112.13.2091
PMID: 10362539
ISSN: 0021-9533
eISSN: 1477-9137
Grant note: HL42385 / NHLBI NIH HHS
Language: English
Date published: 07/1999
Academic Unit: Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Fraternal Order of Eagles Diabetes Research Center; Neurosurgery; Internal Medicine
Record Identifier: 9984020623402771

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