Abstract

Many human diseases are caused by premature translation termination mutations. Because of the prevalence of this class of mutation, a mechanism that prevented translation termination at premature stop mutations would have the potential to reduce or eliminate the disease of a subset of individuals afflicted with a wide range of genetic diseases, including cancer, diabetes, Tay Sachs, beta-thalassemia, hypercholesterolemia, and many others. Another example is the disease cystic fibrosis (CF). CF is caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR). While the most common mutation in the CFTR gene is the deletion of a phenylalanine at residue 508 (delta F508), roughly 10 percent of all CF patients carry at least one premature stop mutation. In preliminary studies, we obtained evidence that premature stop mutations found in CF patients can be suppressed by aminoglycoside antibiotics, resulting in the expression of full-length, functional CFTR. These results suggest that a clinical strategy utilizing aminoglycoside treatment may restore CFTR function in CF patients with this class of mutation. However, it is important that we first understand in greater detail the mechanism of translation termination and how aminoglycosides act to subvert this process. To explore the feasibility of using aminoglycoside-based therapy to suppress disease-causing premature stop mutations, we propose the following Specific Aims:
SPECIFIC AIM 1 : Characterize the aminoglycoside-mediated suppression of premature stop mutations in the CFTR gene.
SPECIFIC AIM 2 : Investigate the ability of aminoglycosides to suppress a naturally-occurring premature stop mutation in the human CFTR gene expressed in a transgenic mouse model.
SPECIFIC AIM 3 : Determine how sequence context affects the efficiency of translation termination in human cells.
SPECIFIC AIM 4 : Characterize the mechanism of translation termination and investigate how aminoglycosides alter this process in mammalian cells.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Specialized Center (P50)
Project #
5P50DK053090-02
Application #
6105821
Study Section
Project Start
1998-09-30
Project End
1999-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
University of Alabama Birmingham
Department
Type
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294

  Publications

Du, Ming; Keeling, Kim M; Fan, Liming et al. (2009) Poly-L-aspartic acid enhances and prolongs gentamicin-mediated suppression of the CFTR-G542X mutation in a cystic fibrosis mouse model. J Biol Chem 284:6885-92
Du, Ming; Liu, Xiaoli; Welch, Ellen M et al. (2008) PTC124 is an orally bioavailable compound that promotes suppression of the human CFTR-G542X nonsense allele in a CF mouse model. Proc Natl Acad Sci U S A 105:2064-9
Guimbellot, Jennifer S; Fortenberry, James A; Siegal, Gene P et al. (2008) Role of oxygen availability in CFTR expression and function. Am J Respir Cell Mol Biol 39:514-21
Berdiev, Bakhrom K; Cormet-Boyaka, Estelle; Tousson, Albert et al. (2007) Molecular proximity of cystic fibrosis transmembrane conductance regulator and epithelial sodium channel assessed by fluorescence resonance energy transfer. J Biol Chem 282:36481-8
Rowe, Steven M; Varga, Karoly; Rab, Andras et al. (2007) Restoration of W1282X CFTR activity by enhanced expression. Am J Respir Cell Mol Biol 37:347-56
Gaggar, Amit; Li, Yao; Weathington, Nathaniel et al. (2007) Matrix metalloprotease-9 dysregulation in lower airway secretions of cystic fibrosis patients. Am J Physiol Lung Cell Mol Physiol 293:L96-L104
Benos, Dale J; Bashari, Edlira; Chaves, Jose M et al. (2007) The ups and downs of peer review. Adv Physiol Educ 31:145-52
Kellermayer, Richard; Szigeti, Reka; Keeling, Kim M et al. (2006) Aminoglycosides as potential pharmacogenetic agents in the treatment of Hailey-Hailey disease. J Invest Dermatol 126:229-31
Su, Xuefeng; Li, Qingnan; Shrestha, Kedar et al. (2006) Interregulation of proton-gated Na(+) channel 3 and cystic fibrosis transmembrane conductance regulator. J Biol Chem 281:36960-8
Du, Ming; Keeling, Kim M; Fan, Liming et al. (2006) Clinical doses of amikacin provide more effective suppression of the human CFTR-G542X stop mutation than gentamicin in a transgenic CF mouse model. J Mol Med 84:573-82

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