Role of actin filament organization in CFTR activation.
The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion-selective channel whose dysfunction leads to the onset of cystic fibrosis. CFTR activation is normally elicited by stimulation of the cAMP pathway, which effects protein kinase A activation. However, previous studies from our laboratory indicate that the actin cytoskeleton is also required for a proper CFTR function. In this report, the regulatory role of actin filament organization in the activation of CFTR was explored. Maneuvers to modify the steady-state organization of actin filaments elicit the activation of CFTR in the absence of a functional cAMP pathway. Partial disruption of the actin cytoskeleton of CFTR-expressing cells with cytochalasin D (CD) induced CFTR activation in the absence of an activated PKA. Similar findings were obtained by intracellular dialysis with the actin-severing protein gelsolin. However, extended treatment with CD leading to the collapse of the actin cytoskeleton rendered CFTR completely insensitive to direct PKA activation. cAMP activation of CFTR was also found to be dysfunctional in cells lacking the actin-crosslinking protein ABP-280, which was recovered after dialysis of the cells with filamin, a homologue of ABP-280. The present data indicate that an organized actin network is required for the proper cAMP-dependent activation of CFTR. The possibility is also explored that actin must be directly associated with CFTR to elicit its activation, further suggesting that this channel protein may bind actin as well.
Cantiello
H F
HF
Renal unit, Massachusetts General Hospital and Department of Medicine, Harvard Medical School, Charlestown, MA 02129, USA. cantiello@helix.mgh.harvard.edu
D008841:Actin Cytoskeleton / Q000378:metabolism*; D000230:Adenocarcinoma; D000595:Amino Acid Sequence; D000818:Animals; D003285:Contractile Proteins / Q000737:chemistry / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000378:metabolism*; D015638:Cytochalasin D / Q000494:pharmacology; D005260:Female; D064448:Filamins; D018260:Gelsolin / Q000378:metabolism; D006801:Humans; D015674:Mammary Neoplasms, Animal; D008545:Melanoma; D051379:Mice; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism; D014407:Tumor Cells, Cultured
C505032:CFTR protein, human; D003285:Contractile Proteins; D064448:Filamins; D018260:Gelsolin; D008840:Microfilament Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D015800:Protozoan Proteins; C036571:severin protein, Dictyostelium; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D015638:Cytochalasin D; D017868:Cyclic AMP-Dependent Protein Kinases