[go: up one dir, main page]
More Web Proxy on the site http://driver.im/
Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Suppression of a CFTR premature stop mutation in a bronchial epithelial cell line

Nature Medicine volume 3, pages 1280–1284 (1997)Cite this article

Abstract

Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR) protein1. While 70% of CF chromosomes carry a deletion of the phenylalanine residue 508 (δF508) of CFTR, roughly 5% of all CF chromosomes carry a premature stop mutation2. We reported that the aminoglycoside antibiotics G-418 and gentamicin can suppress two premature stop mutations [a stop codon in place of glycine residue 542 (G542X) and arginine residue 553 (R553X)] when expressed from a CFTR cDNA in HeLa cells3. Suppression resulted in the synthesis of full-length CFTR protein and the appearance of a cAMP-activated anion conductance characteristic of CFTR function. However, it was unclear whether this approach could restore CFTR function in cells expressing mutant forms of CFTR from the nuclear genome. We now report that C-418 and gentamicin are also capable of restoring CFTR expression in a CF bronchial epithelial cell line carrying the CFTR W1282X premature stop mutation (a stop codon in place of tryptophan residue 1282). This conclusion is based on the reappearance of cAMP-activated chloride currents, the restoration of CFTR protein at the apical plasma membrane, and an increase in the abundance of CFTR mRNA levels from the W1282X allele.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Kerem, B.-S. et al. Identification of the cystic fibrosis gene: Genetic analysis. Science 245, 1073–1080 (1989).

    Article  CAS  Google Scholar 

  2. Population variation of common cystic fibrosis mutations. Hum. Mutat. 4, 167–177 (1994).

  3. Howard, M., Frizzell, R.A. & Bedwell, D.M. Aminoglycoside antibiotics restore CFTR function by overcoming premature stop mutations. Nature Med. 2, 467–469 (1996).

    Article  CAS  Google Scholar 

  4. Bonetti, B., Fu, L., Moon, J. & Bedwell, D.M. The efficiency of translation termination is determined by a synergistic interplay between upstream and downstream sequences in Saccharomyces cerevisiae. J. Mol. Biol. 251, 334–345 (1995).

    Article  CAS  Google Scholar 

  5. Yang, Y. et al. Molecular basis of defective anion transport in L cells. Hum. Mol. Genet. 2, 1253–1261 (1993).

    Article  CAS  Google Scholar 

  6. Zeitlin, P.L. et al. A cystic fibrosis bronchial epithelial cell line: Immortalization by adeno-12 SV40 infection. Am. J. Respir. Cell Mol. Biol. 4, 313–319 (1991).

    Article  CAS  Google Scholar 

  7. Schwiebert, E.M., Flotte, T., Cutting, G.R. & Guggino, W.B. Both CFTR and outwardly rectifying chloride channels contribute to cAMP-stimulated whole cell chloride currents. Am. J. Physiol. 266, C1464–1477 (1994).

    Article  CAS  Google Scholar 

  8. Gabriel, S.E., Clarke, L.L., Boucher, R.C. & Stutts, M.J. CFTR and outwardly rectifying chloride channels are distinct proteins with a regulatory relationship. Nature 363, 263–266 (1993).

    Article  CAS  Google Scholar 

  9. Egan, M.E. et al. Defective regulation of outwardly rectifying chloride channels by protein kinase A corrected by insertion of CFTR. Nature 358, 581–584 (1992).

    Article  CAS  Google Scholar 

  10. Schoumacher, R. et al. A cystic fibrosis pancreatic adenocarcinoma cell line. Proc. Natl. Acad. Sci, USA 87, 4012–4016 (1990).

    Article  CAS  Google Scholar 

  11. Bradbury, N. et al. Regulation of plasma membrane recycling by CFTR. Science 256, 530–532 (1992).

    Article  CAS  Google Scholar 

  12. Prince, L.S., Workman, R.B. & Marchase, R.B. Rapid endocytosis of the cystic fibrosis transmembrane conductance regulator chloride channel. Proc. Natl. Acad. Sci. USA 91, 5192–5196 (1994).

    Article  CAS  Google Scholar 

  13. Tousson, A., Fuller, C.M. & Benos, D.J. Apical recruitment of CFTR in T-84 cells is dependent on cAMP and microtubules but not calcium or microfilaments. J. Cell Sci. 109, 1325–1334 (1996).

    CAS  PubMed  Google Scholar 

  14. Cheng, S.H. et al. Defective intracellular transport and processing is the molecular basis of most cystic fibrosis. Cell 63, 827–834 (1990).

    Article  CAS  Google Scholar 

  15. Hamosh, A., Rosenstein, B.J. & Cutting, G. CFTR nonsense mutations G542X and W1282X associated with severe reduction of CFTR mRNA in nasal epithelial cells. Hum. Mol. Genet. 1, 542–544 (1992).

    Article  CAS  Google Scholar 

  16. Fuller, C.M., Howard, M., Bedwell, D.M., Frizzell, R.A. & Benos, D.J. Antibodies against the cystic fibrosis transmembrane regulator. Am. J. Physiol. 262, C396–C404 (1992).

    Article  CAS  Google Scholar 

  17. Demolombe, S. et al. Abnormal subcellular localization of mutated CFTR protein in a cystic fibrosis epithelial cell line. Eur. J. Cell Biol. 65, 214–219 (1994).

    CAS  PubMed  Google Scholar 

  18. Hamosh, A. et al. Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis. J. Clin. Invest. 88, 1880–1885 (1991).

    Article  CAS  Google Scholar 

  19. Shoshani, T. et al. Association of a nonsense mutation (Wl 282X), the most common mutation in the Ashkenazi Jewish cystic fibrosis patients in Israel, with presentation of severe disease. Am. J. Hum. Cenet. 50, 222–228 (1992).

    CAS  Google Scholar 

  20. Hasty, P. et al. A severe phenotype in mice with a termination mutation in exon 2 of the cystic fibrosis gene. Somat. Cell Mol. Genet. 21, 177–187 (1995).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbiology, The University of Alabama at Birmingham, Bevill Biomedical Research Building, Room 432, 845 Nineteenth Street, South, Birmingham, Alabama, 35294–2170, USA

    David M. Bedwell & Anisa Kaenjak

  2. Department of Physiology and Biophysics, The University of Alabama at Birmingham, Basic Health Sciences Building, Room 706,1918 University Boulevard, Birmingham, Alabama, 35294–0005, USA

    Dale J. Benos, Zsuzsa Bebok, James K. Bubien, J.P. Clancy & Eric J. Sorscher

  3. Department of Cell Biology, The University of Alabama at Birmingham, Basic Health Sciences Building, Room 690,1918 University Boulevard, Birmingham, Alabama, 35294–0005, USA

    Jeong Hong & Albert Tousson

  4. Department of Medicine, The University of Alabama at Birmingham, Basic Health Sciences Building, Room 760, 1918 University Boulevard, Birmingham, Alabama, 35294–0005, USA

    J.P. Clancy & Eric J. Sorscher

  5. Gregory Fleming James Cystic Fibrosis Research Center, The University of Alabama at Birmingham, Basic Health Sciences Building, Room 760, 1918 University Boulevard, Birmingham, Alabama, 35294–0005, USA

    David M. Bedwell, Anisa Kaenjak, Dale J. Benos, Zsuzsa Bebok, James K. Bubien, Jeong Hong, Albert Tousson, J.P. Clancy & Eric J. Sorscher

  6. Department of Medicine, The University of Alabama at Birmingham, Basic Health Sciences Building, Room 690, 1918 University Boulevard, Birmingham, Alabama, 35294–0005, USA

    J.P. Clancy & Eric J. Sorscher

Authors
  1. David M. Bedwell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anisa Kaenjak
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dale J. Benos
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zsuzsa Bebok
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. James K. Bubien
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jeong Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Albert Tousson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J.P. Clancy
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Eric J. Sorscher
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bedwell, D., Kaenjak, A., Benos, D. et al. Suppression of a CFTR premature stop mutation in a bronchial epithelial cell line. Nat Med 3, 1280–1284 (1997). https://doi.org/10.1038/nm1197-1280

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1197-1280

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing