Role of exchange protein activated by cAMP 1 in regulating rates of microtubule formation in cystic fibrosis epithelial cells

Sharon M. Rymut, Tracy Ivy, Deborah A. Corey, Calvin U. Cotton, James D. Burgess, Thomas J. Kelley

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The regulation of microtubule dynamics in cystic fibrosis (CF) epithelial cells and the consequences of reduced rates of microtubule polymerization on downstream CF cellular events, such as cholesterol accumulation, a marker of impaired intracellular transport, are explored here. It is identified that microtubules in both CF cell models and in primary CF nasal epithelial cells repolymerize at a slower rate compared with respective controls. Previous studies suggest a role forcAMPin modulating organelle transport in CF cells, implicating a role for exchange protein activated by cAMP (EPAC) 1, a regulator of microtubule elongation, as a potential mechanism. EPAC1 activity is reduced in CF cell models and in Cftr2/2 mouse lung compared with respective non-CF controls. Stimulation of EPAC1 activity with the selective EPAC1 agonist, 8-cpt-2-O-MecAMP, stimulates microtubule repolymerization to wild-type rates in CF cells. EPAC1 activation also alleviates cholesterol accumulation in CF cells, suggesting a direct link between microtubule regulation and intracellular transport. To verify the relationship between transport and microtubule regulation, expression of the protein, tubulin polymerization-promoting protein, was knocked down in non-CF human tracheal (9/HTEo2) cells to mimic the microtubule dysregulation in CF cells. Transduced cells with short hairpin RNA targeting tubulin polymerization-promoting protein exhibit CF-like perinuclear cholesterol accumulation and other cellular manifestations of CF cells, thus supporting a role for microtubule regulation as a mechanism linking CFTR function to downstream cellular manifestation.

Original languageEnglish (US)
Pages (from-to)853-862
Number of pages10
JournalAmerican journal of respiratory cell and molecular biology
Volume53
Issue number6
DOIs
StatePublished - Dec 2015

Fingerprint

Cystic Fibrosis
Microtubules
Epithelial Cells
Cholesterol
Polymerization
Tubulin
Proteins
Small Interfering RNA
Elongation
Chemical activation
Fibrosis
Nose
Organelles
Lung

Keywords

  • Cholesterol
  • Cystic fibrosis
  • Exchange protein activated by camp 1
  • Microtubules
  • Rap1

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Role of exchange protein activated by cAMP 1 in regulating rates of microtubule formation in cystic fibrosis epithelial cells. / Rymut, Sharon M.; Ivy, Tracy; Corey, Deborah A.; Cotton, Calvin U.; Burgess, James D.; Kelley, Thomas J.

In: American journal of respiratory cell and molecular biology, Vol. 53, No. 6, 12.2015, p. 853-862.

Research output: Contribution to journalArticle

Rymut, Sharon M. ; Ivy, Tracy ; Corey, Deborah A. ; Cotton, Calvin U. ; Burgess, James D. ; Kelley, Thomas J. / Role of exchange protein activated by cAMP 1 in regulating rates of microtubule formation in cystic fibrosis epithelial cells. In: American journal of respiratory cell and molecular biology. 2015 ; Vol. 53, No. 6. pp. 853-862.
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