Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites

Xiu-Bao D Chang, Joseph A. Tabcharani, Yue Xian Hou, Timothy J. Jensen, Norbert Kartner, Noa Alon, John W. Hanrahan, John R. Riordan

Research output: Contribution to journalArticle

241 Citations (Scopus)

Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) plays a central role in transepithelial ion transport by acting as a tightly regulated apical chloride channel. Regulation is achieved by the concerted action of ATP at conserved nucleotide binding folds and serine phosphorylation at multiple sites by protein kinases A (PKA) and C (PKC). A previous investigation concluded that activation by PKA is critically dependent on phosphorylation at four of the nine predicted PKA sites in the R domain (S660A, S737A, S795A, S813A), because a "Quad" mutant lacking these sites could not be activated. We show in the present work that not only can this mutant be phosphorylated and activated, but a mutant in which all 10 predicted PKA sites have been altered still retains significant PKA-activated function. Potentiation of the PKA response by PKC is also preserved in this mutant. Thus CFTR may be regulated by cryptic PKA sites which also mediate interactions between different kinases. Such hierarchical phosphorylation of CFTR by obvious and cryptic PKA sites could provide a metered response to secretagogues. * This work was supported by grants from the Canadian and American Cystic Fibrosis Foundation and the National Institutes of Health-National Institute of Diabetes and Digestive and Kidney Diseases. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Original languageEnglish (US)
Pages (from-to)11304-11311
Number of pages8
JournalJournal of Biological Chemistry
Volume268
Issue number15
StatePublished - May 25 1993
Externally publishedYes

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Cystic Fibrosis Transmembrane Conductance Regulator
Mutagenesis
Chloride Channels
Phosphorylation
Protein Kinases
National Institute of Diabetes and Digestive and Kidney Diseases (U.S.)
Organized Financing
Ion Transport
National Institutes of Health (U.S.)
Medical problems
Cyclic AMP-Dependent Protein Kinases
Cystic Fibrosis
Serine
Protein Kinase C
Publications
Phosphotransferases
Nucleotides
Adenosine Triphosphate
Chemical activation
Health

ASJC Scopus subject areas

  • Biochemistry

Cite this

Chang, X-B. D., Tabcharani, J. A., Hou, Y. X., Jensen, T. J., Kartner, N., Alon, N., ... Riordan, J. R. (1993). Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. Journal of Biological Chemistry, 268(15), 11304-11311.

Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. / Chang, Xiu-Bao D; Tabcharani, Joseph A.; Hou, Yue Xian; Jensen, Timothy J.; Kartner, Norbert; Alon, Noa; Hanrahan, John W.; Riordan, John R.

In: Journal of Biological Chemistry, Vol. 268, No. 15, 25.05.1993, p. 11304-11311.

Research output: Contribution to journalArticle

Chang, X-BD, Tabcharani, JA, Hou, YX, Jensen, TJ, Kartner, N, Alon, N, Hanrahan, JW & Riordan, JR 1993, 'Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites', Journal of Biological Chemistry, vol. 268, no. 15, pp. 11304-11311.
Chang X-BD, Tabcharani JA, Hou YX, Jensen TJ, Kartner N, Alon N et al. Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. Journal of Biological Chemistry. 1993 May 25;268(15):11304-11311.
Chang, Xiu-Bao D ; Tabcharani, Joseph A. ; Hou, Yue Xian ; Jensen, Timothy J. ; Kartner, Norbert ; Alon, Noa ; Hanrahan, John W. ; Riordan, John R. / Protein kinase a (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 15. pp. 11304-11311.
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abstract = "The cystic fibrosis transmembrane conductance regulator (CFTR) plays a central role in transepithelial ion transport by acting as a tightly regulated apical chloride channel. Regulation is achieved by the concerted action of ATP at conserved nucleotide binding folds and serine phosphorylation at multiple sites by protein kinases A (PKA) and C (PKC). A previous investigation concluded that activation by PKA is critically dependent on phosphorylation at four of the nine predicted PKA sites in the R domain (S660A, S737A, S795A, S813A), because a {"}Quad{"} mutant lacking these sites could not be activated. We show in the present work that not only can this mutant be phosphorylated and activated, but a mutant in which all 10 predicted PKA sites have been altered still retains significant PKA-activated function. Potentiation of the PKA response by PKC is also preserved in this mutant. Thus CFTR may be regulated by cryptic PKA sites which also mediate interactions between different kinases. Such hierarchical phosphorylation of CFTR by obvious and cryptic PKA sites could provide a metered response to secretagogues. * This work was supported by grants from the Canadian and American Cystic Fibrosis Foundation and the National Institutes of Health-National Institute of Diabetes and Digestive and Kidney Diseases. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked {"}advertisement{"} in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.",
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