Cystic fibrosis (CF) is one of the most common autosomal recessive diseases of man, affecting 1 in 2500 Caucasian newborns. The disease primarily affects the lungs and pancreas, where damage results from the thick sticky mucus which accumulates and leads to obstruction and organ damage. The current average survival is about age 25, with death usually coming from respiratory failure. After years of work, the CF gene was cloned by our laboratory in collaboration with the Hospital for Sick Children in Toronto in 1989, using the """"""""reverse genetics"""""""" strategy. The gene encodes a 1480 amino acid membrane protein, and the common mutation is a deletion of phenylalanine 508. We now propose a series of studies to further define the function and regulation of expression of the CF gene. This will include the identification of additional mutations responsible for CF by a variety of sensitive techniques, a study of the function of the gene by transfection into cystic fibrosis cells and assessment of correction of the defect, site-specific mutagenesis of various regions of the gene, the generation of antibodies to domains of the protein product in order to further characterize its expression, and a study of the regulation of transcription by defining the promoter sequences responsible for epithelial specificity. Furthermore, we have recently identified evidence for alternative splicing of the gene which is likely to have important functional consequences, and this phenomenon will be intensively investigated. These studies should bring us much closer to an understanding of the normal function of the CF gene, and should lay the groundwork for the development of effective therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK039690-06
Application #
3239584
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1988-01-01
Project End
1993-04-05
Budget Start
1993-01-01
Budget End
1993-04-05
Support Year
6
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Wilkinson, D J; Strong, T V; Mansoura, M K et al. (1997) CFTR activation: additive effects of stimulatory and inhibitory phosphorylation sites in the R domain. Am J Physiol 273:L127-33
Strong, T V; Boehm, K; Collins, F S (1994) Localization of cystic fibrosis transmembrane conductance regulator mRNA in the human gastrointestinal tract by in situ hybridization. J Clin Invest 93:347-54
Cohn, J A; Strong, T V; Picciotto, M R et al. (1993) Localization of the cystic fibrosis transmembrane conductance regulator in human bile duct epithelial cells. Gastroenterology 105:1857-64
Strong, T V; Wilkinson, D J; Mansoura, M K et al. (1993) Expression of an abundant alternatively spliced form of the cystic fibrosis transmembrane conductance regulator (CFTR) gene is not associated with a cAMP-activated chloride conductance. Hum Mol Genet 2:225-30
Sferra, T J; Collins, F S (1993) The molecular biology of cystic fibrosis. Annu Rev Med 44:133-44
Smit, L S; Nasr, S Z; Iannuzzi, M C et al. (1993) An African-American cystic fibrosis patient homozygous for a novel frameshift mutation associated with reduced CFTR mRNA levels. Hum Mutat 2:148-51
Smit, L S; Wilkinson, D J; Mansoura, M K et al. (1993) Functional roles of the nucleotide-binding folds in the activation of the cystic fibrosis transmembrane conductance regulator. Proc Natl Acad Sci U S A 90:9963-7
Koh, J; Sferra, T J; Collins, F S (1993) Characterization of the cystic fibrosis transmembrane conductance regulator promoter region. Chromatin context and tissue-specificity. J Biol Chem 268:15912-21
Collins, F S (1992) Cystic fibrosis: molecular biology and therapeutic implications. Science 256:774-9
Strong, T V; Smit, L S; Nasr, S et al. (1992) Characterization of an intron 12 splice donor mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Hum Mutat 1:380-7

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