The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes
Background/Aims: Cystic Fibrosis (CF) is caused by mutations in the CFTR gene, encoding a cAMP-activated chloride (Cl-) channel. We have previously demonstrated that the expression of several genes can be modulated by the CFTR activity; among them, SRC, MTND4, CISD1, and IL1B. However, the CFTR sign...
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doaj-719819a69df041a19ef8bf3808521a392020-11-24T21:56:16ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782016-01-01381496410.1159/000438608438608The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific GenesÁngel G. ValdiviesoMariángeles ClauzureMacarena Massip-CopizTomás A. Santa-ColomaBackground/Aims: Cystic Fibrosis (CF) is caused by mutations in the CFTR gene, encoding a cAMP-activated chloride (Cl-) channel. We have previously demonstrated that the expression of several genes can be modulated by the CFTR activity; among them, SRC, MTND4, CISD1, and IL1B. However, the CFTR signalling mechanism involved in the expression of CFTR-dependent genes is unknown. The aim of this work was to determine if intracellular chloride (Cl-)i might function as a second messenger modulating the expression of specific genes. Methods: Differential display (DD) was applied to IB3-1 cells (CF cells), cultured under conditions that produce different intracellular Cl- concentrations ([Cl-]i), to analyse their expression profile. Results: Several differentially expressed gene products were observed by using DD, suggesting the presence of chloride-dependent gene expression. Two cDNA fragments, derived from differentially expressed mRNAs and showing opposed response to Cl-‚ were isolated, cloned, sequenced and its Cl- dependency validated by reverse transcription quantitative-PCR (RT-qPCR). We identified the gene RPS27, which encodes the multifunctional ribosomal protein RPS27, also known as metallopanstimulin-1 (MPS-1), and the gene GLRX5, encoding glutaredoxin-related protein 5, as chloride-dependent genes. RPS27 was negatively regulated with increased [Cl-]i, approximately from 25-75 mM Cl- (EC50 = 46 ± 7 mM), and positively regulated from 75-125 mM Cl- (EC50 = 110 ± 11 mM) (biphasic response). In contrast, GLRX5 was positively modulated by [Cl-]i, showing a typical sigmoidal dose-response curve from 0-50 mM Cl-, reaching a plateau after 50 mM Cl- (EC50 ∼ 34 mM). Conclusion: The results suggest the existence of chloride-dependent genes. The Cl- anion, therefore, might act as a second messenger for channels or receptors able to modulate the intracellular Cl- concentration, regulating in turn the expression of specific genes.http://www.karger.com/Article/FullText/438608GLRX5RPS27Chloride-dependent gene expressionIntracellular chlorideSecond messenger |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ángel G. Valdivieso Mariángeles Clauzure Macarena Massip-Copiz Tomás A. Santa-Coloma |
spellingShingle |
Ángel G. Valdivieso Mariángeles Clauzure Macarena Massip-Copiz Tomás A. Santa-Coloma The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes Cellular Physiology and Biochemistry GLRX5 RPS27 Chloride-dependent gene expression Intracellular chloride Second messenger |
author_facet |
Ángel G. Valdivieso Mariángeles Clauzure Macarena Massip-Copiz Tomás A. Santa-Coloma |
author_sort |
Ángel G. Valdivieso |
title |
The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes |
title_short |
The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes |
title_full |
The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes |
title_fullStr |
The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes |
title_full_unstemmed |
The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes |
title_sort |
chloride anion acts as a second messenger in mammalian cells - modifying the expression of specific genes |
publisher |
Cell Physiol Biochem Press GmbH & Co KG |
series |
Cellular Physiology and Biochemistry |
issn |
1015-8987 1421-9778 |
publishDate |
2016-01-01 |
description |
Background/Aims: Cystic Fibrosis (CF) is caused by mutations in the CFTR gene, encoding a cAMP-activated chloride (Cl-) channel. We have previously demonstrated that the expression of several genes can be modulated by the CFTR activity; among them, SRC, MTND4, CISD1, and IL1B. However, the CFTR signalling mechanism involved in the expression of CFTR-dependent genes is unknown. The aim of this work was to determine if intracellular chloride (Cl-)i might function as a second messenger modulating the expression of specific genes. Methods: Differential display (DD) was applied to IB3-1 cells (CF cells), cultured under conditions that produce different intracellular Cl- concentrations ([Cl-]i), to analyse their expression profile. Results: Several differentially expressed gene products were observed by using DD, suggesting the presence of chloride-dependent gene expression. Two cDNA fragments, derived from differentially expressed mRNAs and showing opposed response to Cl-‚ were isolated, cloned, sequenced and its Cl- dependency validated by reverse transcription quantitative-PCR (RT-qPCR). We identified the gene RPS27, which encodes the multifunctional ribosomal protein RPS27, also known as metallopanstimulin-1 (MPS-1), and the gene GLRX5, encoding glutaredoxin-related protein 5, as chloride-dependent genes. RPS27 was negatively regulated with increased [Cl-]i, approximately from 25-75 mM Cl- (EC50 = 46 ± 7 mM), and positively regulated from 75-125 mM Cl- (EC50 = 110 ± 11 mM) (biphasic response). In contrast, GLRX5 was positively modulated by [Cl-]i, showing a typical sigmoidal dose-response curve from 0-50 mM Cl-, reaching a plateau after 50 mM Cl- (EC50 ∼ 34 mM). Conclusion: The results suggest the existence of chloride-dependent genes. The Cl- anion, therefore, might act as a second messenger for channels or receptors able to modulate the intracellular Cl- concentration, regulating in turn the expression of specific genes. |
topic |
GLRX5 RPS27 Chloride-dependent gene expression Intracellular chloride Second messenger |
url |
http://www.karger.com/Article/FullText/438608 |
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