VCP/p97 AAA-ATPase does not interact with the endogenous wild-type cystic fibrosis transmembrane conductance regulator

被引:11
作者
Goldstein, Rebecca F.
Niraj, Ashutosh
Sanderson, Todd P.
Wilson, Landon S.
Rab, Andras
Kim, Helen
Bebok, Zsuzsa
Collawn, James F.
机构
[1] Univ Alabama, Dept Cell Biol, Dept Pharmacol & Toxicol, Ctr Comprehens Canc, Birmingham, AL 35294 USA
[2] Univ Alabama, Gregory Fleming James Cyst Fibrosis Ctr, Birmingham, AL 35294 USA
关键词
CFTR; p97/VCP; epithelium; ubiquitination; biogenesis;
D O I
10.1165/rcmb.2006-0365OC
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel that is defective in cystic fibrosis. The most common mutation, Delta F508 CFTR, is retained in the endoplasmic reticulum, retrotranslocated into the cytosol, and degraded by the proteasome. In a proteomics screen to identify Delta F508 CFTR interacting proteins, we found that valosin-containing protein (VCP)/ p97, a Type II AAA ATPase that is a component of the retrotranslocation machinery, binds Delta F508 CFTR, and this interaction is stabilized by proteasomal inhibition. Since wild-type (WT) CFTR has been reported to be inefficiently processed during biogenesis with as much as 75% of the newly synthesized protein degraded by the proteasome, we examined the VCP interaction in Calu-3, T-84, and 16HBE, three epithelial cell lines that endogenously express WT CFTR. The results indicate that when WT CFTR processing is efficient, as demonstrated in Calu-3 cells, VCP does not interact. Interestingly, overexpression of recombinant WT CFTR in Calu-3 cells results in inefficient processing and VCP interaction, demonstrating that CFTR processing efficiency and the VCP interaction are tightly coupled. Furthermore, induction of ER stress and activation of the unfolded protein response result in inefficient processing of WT CFTR in Calu-3 cells and promote the WT CFTR-VCP interaction. The results support the hypothesis that components of the retrotranslocation machinery such as VCP do not interact with CFTR in epithelial cells that endogenously express WT CFTR, since under normal conditions the processing of the WT protein is efficient.
引用
收藏
页码:706 / 714
页数:9
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