title: RACK1 interacts with filamin-A to regulate plasma membrane levels of the cystic fibrosis transmembrane conductance regulator.
abstract: Mutations in cystic fibrosis transmembrane regulator (CFTR), a chloride channel in the apical membranes of secretory epithelial cells, underlie the fatal genetic disorder cystic fibrosis. Certain CFTR mutations, including the common mutation ΔF508-CFTR, result in greatly decreased levels of active CFTR at the apical membrane. Direct interactions between CFTR and the cytoskeletal adaptors filamin-A (FlnA) and Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) stabilize the expression and localization of CFTR at the plasma membrane. The scaffold protein receptor for activated C kinase 1 (RACK1) also stabilizes CFTR surface expression; however, RACK1 does not interact directly with CFTR and its mechanism of action is unknown. In the present study, we report that RACK1 interacts directly with FlnA in vitro and in a Calu-3 airway epithelial cell line. We mapped the interaction between RACK1 and FlnA to the WD4 and WD6 repeats of RACK1 and to a segment of the large rod domain of FlnA, consisting of immunoglobulin-like repeats 8-15. Disruption of the RACK1-FlnA interaction causes a reduction in CFTR surface levels. Our results suggest that a novel RACK1-FlnA interaction is an important regulator of CFTR surface localization.
authors: Willard Alan Bernbaum Center for Cystic Fibrosis Research, Case Western Reserve University, Cleveland, OH 44106, USA.
authors: Laura
authors: L
authors: Smith
authors: Paul
authors: P
authors: Litman
authors: Ekta
authors: E
authors: Kohli
authors: Joseph
authors: J
authors: Amick
authors: Richard C
authors: RC
authors: Page
authors: Saurav
authors: S
authors: Misra
authors: Carole M
authors: CM
authors: Liedtke
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D003285:Contractile Proteins / Q000235:genetics / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D064448:Filamins; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport / Q000502:physiology; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism*
chemical_list: C505032:CFTR protein, human; D003285:Contractile Proteins; D064448:Filamins; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019204:GTP-Binding Proteins
keywords: Ig repeat; WD repeat; cystic fibrosis transmembrane regulator; β-propeller
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