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Related Concept Videos

Cystic Fibrosis: Pathogenesis01:23

Cystic Fibrosis: Pathogenesis

Cystic fibrosis (CF), an autosomal recessive disorder, significantly affects the function of exocrine glands. This genetically inherited disease is characterized by the production of thick and sticky mucus, which can severely affect various organs and systems in the body.
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Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
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Updated: May 25, 2026

In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes
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Phosphorylation-dependent 14-3-3 protein interactions regulate CFTR biogenesis.

Xiubin Liang1, Ana Carina Da Paula, Zoltán Bozóky

  • 1Department of Cell Biology and Physiology, University of Pittsburgh, Pittsburgh, PA 15261, USA.

Molecular Biology of the Cell
|January 27, 2012
PubMed
Summary

The study shows that 14-3-3 proteins enhance cystic fibrosis transmembrane conductance regulator (CFTR) biogenesis by binding to phosphorylated CFTR, increasing its cell surface presence and anion secretion. This mechanism is regulated by cAMP/protein kinase A (PKA) signaling.

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In Vitro Analysis of PDZ-dependent CFTR Macromolecular Signaling Complexes
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Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae
14:56

Expression and Purification of the Cystic Fibrosis Transmembrane Conductance Regulator Protein in Saccharomyces cerevisiae

Published on: March 10, 2012

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Physiology

Background:

  • Cystic fibrosis transmembrane conductance regulator (CFTR) is a critical chloride channel regulated by cAMP/protein kinase A (PKA) phosphorylation.
  • CFTR phosphorylation controls anion secretion across epithelial cell apical membranes.
  • The posttranslational regulation of CFTR biogenesis by cAMP/PKA signaling remains incompletely understood.

Purpose of the Study:

  • To investigate the hypothesis that cAMP/PKA stimulation regulates CFTR biogenesis posttranslationally.
  • To elucidate the role of 14-3-3 proteins in CFTR trafficking and function.
  • To determine how 14-3-3 binding to CFTR influences anion secretion.

Main Methods:

  • Coimmunoprecipitation assays to assess protein interactions.
  • Forskolin stimulation to mimic cAMP/PKA activation.
  • Pulse-chase experiments to track CFTR synthesis and degradation.
  • Nuclear magnetic resonance (NMR) analysis to determine binding sites.
  • Assessment of cell surface CFTR levels and anion efflux.

Main Results:

  • 14-3-3β, γ, and ε isoforms were expressed in airway cells and interacted with CFTR.
  • Forskolin stimulation increased 14-3-3β and ε binding to CFTR.
  • Overexpression of 14-3-3 proteins increased CFTR biogenesis and cell surface expression.
  • 14-3-3β knockdown reduced CFTR levels, cell surface expression, and anion efflux.
  • Forskolin stimulated CFTR/14-3-3β interaction and reduced CFTR interaction with COP1.

Conclusions:

  • 14-3-3 proteins bind to phosphorylated CFTR at PKA-regulated sites.
  • 14-3-3 binding augments CFTR biogenesis by reducing endoplasmic reticulum retrieval.
  • cAMP/PKA stimulation enhances CFTR availability for anion secretion via 14-3-3 interaction.