The cystic-fibrosis-associated ΔF508 mutation confers post-transcriptional destabilization on the C. elegans ABC transporter PGP-3

• Published in: Disease models & mechanisms 2012-11, Vol.5 (6), p.930-939
• Main Authors: He, Liping, Skirkanich, Jennifer, Moronetti, Lorenza, Lewis, Rosemary, Lamitina, Todd
• Format: Article
• Language: English
• Subjects: ABC transporters; Amino acids; Animals; ATP Binding Cassette Transporter, Sub-Family B - genetics; ATP Binding Cassette Transporter, Sub-Family B - metabolism; ATP-Binding Cassette Transporters - genetics; ATP-Binding Cassette Transporters - metabolism; Caenorhabditis elegans - cytology; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Cell Membrane - metabolism; Cell Polarity; Cystic fibrosis; Cystic Fibrosis - genetics; Cystic Fibrosis Transmembrane Conductance Regulator - genetics; Endoplasmic Reticulum-Associated Degradation; Enzymes; Gene Knockdown Techniques; Genomes; Humans; Mammals; Mutation; Mutation - genetics; Osmotic Pressure; Plasma; Protein folding; Protein Stability; Quality control; Research Report; Stress, Physiological - genetics; Transcription, Genetic; Worms
• ISSN: 1754-8403; 1754-8411
• DOI: 10.1242/dmm.008987

Membrane proteins make up ∼30% of the proteome. During the early stages of maturation, this class of proteins can experience localized misfolding in distinct cellular compartments, such as the cytoplasm, endoplasmic reticulum (ER) lumen and ER membrane. ER quality control (ERQC) mechanisms monitor folding and determine whether a membrane protein is appropriately folded or is misfolded and warrants degradation. ERQC plays crucial roles in human diseases, such as cystic fibrosis, in which deletion of a single amino acid (F508) results in the misfolding and degradation of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel. We introduced the ΔF508 mutation into Caenorhabditis elegans PGP-3, a 12-transmembrane ABC transporter with 15% identity to CFTR. When expressed in intestinal epithelial cells, PGP-3(wt) was stable and efficiently trafficked to the apical plasma membrane through a COPII-dependent mechanism. However, PGP-3(ΔF508) was post-transcriptionally destabilized, resulting in reduced total and apical membrane protein levels. Genetic or physiological activation of the osmotic stress response pathway, which causes accumulation of the chemical chaperone glycerol, stabilized PGP-3(ΔF508). Efficient degradation of PGP-3(ΔF508) required the function of several C. elegans ER-associated degradation (ERAD) homologs, suggesting that destabilization occurs through an ERAD-type mechanism. Our studies show that the ΔF508 mutation causes post-transcriptional destabilization and degradation of PGP-3 in C. elegans epithelial cells. This model, combined with the power of C. elegans genetics, provides a new opportunity to genetically dissect metazoan ERQC.