TY - JOUR
T1 - Sodium 4-phenylbutyrate downregulates Hsc70
T2 - Implications for intracellular trafficking of ΔF508-CFTR
AU - Rubenstein, Ronald C.
AU - Zeitlin, Pamela L.
PY - 2000
Y1 - 2000
N2 - The most common mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), ΔF508, is a trafficking mutant that has prolonged associations with molecular chaperones and is rapidly degraded, at least in part by the ubiquitin-proteasome system. Sodium 4-phenylbutyrate (4PBA) improves ΔF508-CFTR trafficking and function in vitro in cystic fibrosis epithelial cells and in vivo. To further understand the mechanism of action of 4PBA, we tested the hypothesis that 4PBA modulates the targeting of ΔF508-CFTR for ubiquitination and degradation by reducing the expression of Hsc70 in cystic fibrosis epithelial cells. IB3-1 cells (genotype ΔF508/W1282X) that were treated with 0.05-5 mM 4PBA for 2 days in culture demonstrated a dose-dependent reduction in Hsc70 protein immunoreactivity and mRNA levels. Immunoprecipitation with Hsc70-specific antiserum demonstrated that Hsc70 and CFTR associated under control conditions and that treatment with 4PBA reduced these complexes. Levels of immunoreactive Hsp40, Hdj2, Hsp70, Hsp90, and calnexin were unaffected by 4PBA treatment. These data suggest that 4PBA may improve ΔF508-CFTR trafficking by allowing a greater proportion of mutant CFTR to escape association with Hsc70.
AB - The most common mutation of the cystic fibrosis transmembrane conductance regulator (CFTR), ΔF508, is a trafficking mutant that has prolonged associations with molecular chaperones and is rapidly degraded, at least in part by the ubiquitin-proteasome system. Sodium 4-phenylbutyrate (4PBA) improves ΔF508-CFTR trafficking and function in vitro in cystic fibrosis epithelial cells and in vivo. To further understand the mechanism of action of 4PBA, we tested the hypothesis that 4PBA modulates the targeting of ΔF508-CFTR for ubiquitination and degradation by reducing the expression of Hsc70 in cystic fibrosis epithelial cells. IB3-1 cells (genotype ΔF508/W1282X) that were treated with 0.05-5 mM 4PBA for 2 days in culture demonstrated a dose-dependent reduction in Hsc70 protein immunoreactivity and mRNA levels. Immunoprecipitation with Hsc70-specific antiserum demonstrated that Hsc70 and CFTR associated under control conditions and that treatment with 4PBA reduced these complexes. Levels of immunoreactive Hsp40, Hdj2, Hsp70, Hsp90, and calnexin were unaffected by 4PBA treatment. These data suggest that 4PBA may improve ΔF508-CFTR trafficking by allowing a greater proportion of mutant CFTR to escape association with Hsc70.
KW - Chaperones
KW - Cystic fibrosis
KW - Cystic fibrosis transmembrane conductance regulator
KW - Hsc70
KW - Phenylbutyrate
UR - http://www.scopus.com/inward/record.url?scp=0034099743&partnerID=8YFLogxK
U2 - 10.1152/ajpcell.2000.278.2.c259
DO - 10.1152/ajpcell.2000.278.2.c259
M3 - Article
C2 - 10666020
AN - SCOPUS:0034099743
SN - 0363-6143
VL - 278
SP - C259-C267
JO - American Journal of Physiology - Cell Physiology
JF - American Journal of Physiology - Cell Physiology
IS - 2 47-2
ER -