Abstract

Cystic fibrosis is now known to be due to a recessive genetic defect in the regulation of anion transport in epithelial cells. In these epithelia, chloride is transported across the cell membrane through a channel that can be opened by cyclic AMP dependent protein kinase. Recently, it was shown that in cystic fibrosis, the kinase fails to open the channel, even though the channel is present in the membrane. The Columbia-Presbyterian Cystic Fibrosis Center is composed of a multidisciplinary group of investigators who are collaborating towards identification of the molecular defect in this disease. We have already purified the epithelial chloride channel and using this procedure we will clone and sequence the gene for the channel. Methods have been developed for expression of ion transport proteins in frog oocytes which will facilitate the cloning of the chloride channel. Another expression system was developed which relies on DNA-mediated gene transfer, that will allow the cloning of iodide and chloride transport proteins from thyroid epithelial cells. This method will also be used to identify structural and regulatory mutants of these genes. Regulation of the chloride channel by a variety of protein kineses will be studied in human epithelial cells and in cells derived from patients with cystic fibrosis. The chloride channel will be reconstituted in planar lipid bilayer and the effect of kineses on its electrophysiological characteristics will be studied. The Center will promote the collaboration of all the involved groups towards the identification of the genetic defect in cystic fibrosis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Specialized Center (P50)
Project #
5P50DK041146-06
Application #
3105887
Study Section
Special Emphasis Panel (SRC (TE))
Project Start
1988-09-30
Project End
1994-08-31
Budget Start
1993-09-27
Budget End
1994-08-31
Support Year
6
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027

  Publications

Feliciello, A; Giuliano, P; Porcellini, A et al. (1996) The v-Ki-Ras oncogene alters cAMP nuclear signaling by regulating the location and the expression of cAMP-dependent protein kinase IIbeta. J Biol Chem 271:25350-9
Sullivan, S K; Agellon, L B; Schick, R (1995) Identification and partial characterization of a domain in CFTR that may bind cyclic nucleotides directly. Curr Biol 5:1159-67
Imundo, L; Barasch, J; Prince, A et al. (1995) Cystic fibrosis epithelial cells have a receptor for pathogenic bacteria on their apical surface. Proc Natl Acad Sci U S A 92:3019-23
Gallo, A; Feliciello, A; Varrone, A et al. (1995) Ki-ras oncogene interferes with the expression of cyclic AMP-dependent promoters. Cell Growth Differ 6:91-5
Akabas, M H; Kaufmann, C; Cook, T A et al. (1994) Amino acid residues lining the chloride channel of the cystic fibrosis transmembrane conductance regulator. J Biol Chem 269:14865-8
Landry, D; Sullivan, S; Nicolaides, M et al. (1993) Molecular cloning and characterization of p64, a chloride channel protein from kidney microsomes. J Biol Chem 268:14948-55
Gallo, A; Benusiglio, E; Bonapace, I M et al. (1992) v-ras and protein kinase C dedifferentiate thyroid cells by down-regulating nuclear cAMP-dependent protein kinase A. Genes Dev 6:1621-30
Redhead, C R; Edelman, A E; Brown, D et al. (1992) A ubiquitous 64-kDa protein is a component of a chloride channel of plasma and intracellular membranes. Proc Natl Acad Sci U S A 89:3716-20
Avvedimento, V E; Musti, A M; Ueffing, M et al. (1991) Reversible inhibition of a thyroid-specific trans-acting factor by Ras. Genes Dev 5:22-8
Sullivan, S K; Swamy, K; Field, M (1991) cAMP-activated C1 conductance is expressed in Xenopus oocytes by injection of shark rectal gland mRNA. Am J Physiol 260:C664-9

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