Antisense oligodeoxynucleotides to the cystic fibrosis transmembrane conductance regulator inhibit cAMP-activated but not calcium-activated chloride currents.
Identifieur interne : 000221 ( Ncbi/Checkpoint ); précédent : 000220; suivant : 000222Antisense oligodeoxynucleotides to the cystic fibrosis transmembrane conductance regulator inhibit cAMP-activated but not calcium-activated chloride currents.
Auteurs : J A Wagner [États-Unis] ; T V Mcdonald ; P T Nghiem ; A W Lowe ; H. Schulman ; D C Gruenert ; L. Stryer ; P. GardnerSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1992.
Descripteurs français
- KwdFr :
- AMP cyclique (analogues et dérivés), AMP cyclique (métabolisme), AMP cyclique (pharmacologie), ARN messager (génétique), ARN messager (métabolisme), Calcium (pharmacologie), Canaux chlorure, Canaux ioniques (), Données de séquences moléculaires, Humains, Ionomycine (pharmacologie), Lignée cellulaire, Modèles biologiques, Mucoviscidose (génétique), Oligonucléotides antisens (pharmacologie), Phosphorylation, Protéine CFTR, Protéines membranaires (), Protéines membranaires (génétique), Protéines membranaires (physiologie), Séquence nucléotidique, Thionucléotides (pharmacologie).
- MESH :
- analogues et dérivés : AMP cyclique.
- génétique : ARN messager, Mucoviscidose, Protéines membranaires.
- métabolisme : AMP cyclique, ARN messager.
- pharmacologie : AMP cyclique, Calcium, Ionomycine, Oligonucléotides antisens, Thionucléotides.
- physiologie : Protéines membranaires.
- Canaux chlorure, Canaux ioniques, Données de séquences moléculaires, Humains, Lignée cellulaire, Modèles biologiques, Phosphorylation, Protéine CFTR, Protéines membranaires, Séquence nucléotidique.
English descriptors
- KwdEn :
- Base Sequence, Calcium (pharmacology), Cell Line, Chloride Channels, Cyclic AMP (analogs & derivatives), Cyclic AMP (metabolism), Cyclic AMP (pharmacology), Cystic Fibrosis (genetics), Cystic Fibrosis Transmembrane Conductance Regulator, Humans, Ion Channels (drug effects), Ionomycin (pharmacology), Membrane Proteins (drug effects), Membrane Proteins (genetics), Membrane Proteins (physiology), Models, Biological, Molecular Sequence Data, Oligonucleotides, Antisense (pharmacology), Phosphorylation, RNA, Messenger (genetics), RNA, Messenger (metabolism), Thionucleotides (pharmacology).
- MESH :
- chemical , analogs & derivatives : Cyclic AMP.
- chemical , drug effects : Ion Channels, Membrane Proteins.
- chemical , genetics : Membrane Proteins, RNA, Messenger.
- chemical , metabolism : Cyclic AMP, RNA, Messenger.
- chemical , pharmacology : Calcium, Cyclic AMP, Ionomycin, Oligonucleotides, Antisense, Thionucleotides.
- genetics : Cystic Fibrosis.
- chemical , physiology : Membrane Proteins.
- Base Sequence, Cell Line, Chloride Channels, Cystic Fibrosis Transmembrane Conductance Regulator, Humans, Models, Biological, Molecular Sequence Data, Phosphorylation.
Abstract
Phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR) by cAMP-dependent protein kinase leads to chloride flux in epithelial cells. Is CFTR also required for the calcium-dependent activation of chloride channels? We used antisense oligodeoxynucleotides to CFTR to reduce the expression of CFTR in colonic and tracheal epithelial cells. The antisense oligomers were a pair of adjacent 18-mers complementary to nucleotides 1-18 and 19-36 of CFTR mRNA. Sense and misantisense oligomers served as controls. A 48-h antisense treatment reduced the expression of CFTR protein as assayed by immunoprecipitation and autoradiography to 26% of the level in sense-treated T84 cells. Whole-cell patch clamp revealed that a 48-h antisense treatment of T84 and 56FHTE-8o- fetal tracheal epithelial cells reduced the cAMP-activated chloride current to approximately 10% of that in sense-treated cells. The half-life of functional CFTR is less than 24 h in these cells. In contrast, the calcium-activated chloride current was not affected by antisense treatment. Hence, the cAMP and calcium pathways are separate. CFTR is required for the cAMP pathway but not for the calcium pathway.
DOI: 10.1073/pnas.89.15.6785
PubMed: 1379720
Affiliations:
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pubmed:1379720Le document en format XML
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Schulman</name></author><author><name sortKey="Gruenert, D C" sort="Gruenert, D C" uniqKey="Gruenert D" first="D C" last="Gruenert">D C Gruenert</name></author><author><name sortKey="Stryer, L" sort="Stryer, L" uniqKey="Stryer L" first="L" last="Stryer">L. Stryer</name></author><author><name sortKey="Gardner, P" sort="Gardner, P" uniqKey="Gardner P" first="P" last="Gardner">P. Gardner</name></author></analytic><series><title level="j">Proceedings of the National Academy of Sciences of the United States of America</title><idno type="ISSN">0027-8424</idno><imprint><date when="1992" type="published">1992</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>Calcium (pharmacology)</term><term>Cell Line</term><term>Chloride Channels</term><term>Cyclic AMP (analogs & derivatives)</term><term>Cyclic AMP (metabolism)</term><term>Cyclic AMP (pharmacology)</term><term>Cystic Fibrosis (genetics)</term><term>Cystic Fibrosis Transmembrane Conductance Regulator</term><term>Humans</term><term>Ion Channels (drug effects)</term><term>Ionomycin (pharmacology)</term><term>Membrane Proteins (drug effects)</term><term>Membrane Proteins (genetics)</term><term>Membrane Proteins (physiology)</term><term>Models, Biological</term><term>Molecular Sequence Data</term><term>Oligonucleotides, Antisense (pharmacology)</term><term>Phosphorylation</term><term>RNA, Messenger (genetics)</term><term>RNA, Messenger (metabolism)</term><term>Thionucleotides (pharmacology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>AMP cyclique (analogues et dérivés)</term><term>AMP cyclique (métabolisme)</term><term>AMP cyclique (pharmacologie)</term><term>ARN messager (génétique)</term><term>ARN messager (métabolisme)</term><term>Calcium (pharmacologie)</term><term>Canaux chlorure</term><term>Canaux ioniques ()</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Ionomycine (pharmacologie)</term><term>Lignée cellulaire</term><term>Modèles biologiques</term><term>Mucoviscidose (génétique)</term><term>Oligonucléotides antisens (pharmacologie)</term><term>Phosphorylation</term><term>Protéine CFTR</term><term>Protéines membranaires ()</term><term>Protéines membranaires (génétique)</term><term>Protéines membranaires (physiologie)</term><term>Séquence nucléotidique</term><term>Thionucléotides (pharmacologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Cyclic AMP</term></keywords><keywords scheme="MESH" type="chemical" qualifier="drug effects" xml:lang="en"><term>Ion Channels</term><term>Membrane Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Membrane Proteins</term><term>RNA, Messenger</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cyclic AMP</term><term>RNA, Messenger</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Calcium</term><term>Cyclic AMP</term><term>Ionomycin</term><term>Oligonucleotides, Antisense</term><term>Thionucleotides</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>AMP cyclique</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Cystic Fibrosis</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN messager</term><term>Mucoviscidose</term><term>Protéines membranaires</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>AMP cyclique</term><term>ARN messager</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>AMP cyclique</term><term>Calcium</term><term>Ionomycine</term><term>Oligonucléotides antisens</term><term>Thionucléotides</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Protéines membranaires</term></keywords><keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Membrane Proteins</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Cell Line</term><term>Chloride Channels</term><term>Cystic Fibrosis Transmembrane Conductance Regulator</term><term>Humans</term><term>Models, Biological</term><term>Molecular Sequence Data</term><term>Phosphorylation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Canaux chlorure</term><term>Canaux ioniques</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Lignée cellulaire</term><term>Modèles biologiques</term><term>Phosphorylation</term><term>Protéine CFTR</term><term>Protéines membranaires</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR) by cAMP-dependent protein kinase leads to chloride flux in epithelial cells. Is CFTR also required for the calcium-dependent activation of chloride channels? We used antisense oligodeoxynucleotides to CFTR to reduce the expression of CFTR in colonic and tracheal epithelial cells. The antisense oligomers were a pair of adjacent 18-mers complementary to nucleotides 1-18 and 19-36 of CFTR mRNA. Sense and misantisense oligomers served as controls. A 48-h antisense treatment reduced the expression of CFTR protein as assayed by immunoprecipitation and autoradiography to 26% of the level in sense-treated T84 cells. Whole-cell patch clamp revealed that a 48-h antisense treatment of T84 and 56FHTE-8o- fetal tracheal epithelial cells reduced the cAMP-activated chloride current to approximately 10% of that in sense-treated cells. The half-life of functional CFTR is less than 24 h in these cells. In contrast, the calcium-activated chloride current was not affected by antisense treatment. Hence, the cAMP and calcium pathways are separate. CFTR is required for the cAMP pathway but not for the calcium pathway.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Californie</li></region></list><tree><noCountry><name sortKey="Gardner, P" sort="Gardner, P" uniqKey="Gardner P" first="P" last="Gardner">P. Gardner</name><name sortKey="Gruenert, D C" sort="Gruenert, D C" uniqKey="Gruenert D" first="D C" last="Gruenert">D C Gruenert</name><name sortKey="Lowe, A W" sort="Lowe, A W" uniqKey="Lowe A" first="A W" last="Lowe">A W Lowe</name><name sortKey="Mcdonald, T V" sort="Mcdonald, T V" uniqKey="Mcdonald T" first="T V" last="Mcdonald">T V Mcdonald</name><name sortKey="Nghiem, P T" sort="Nghiem, P T" uniqKey="Nghiem P" first="P T" last="Nghiem">P T Nghiem</name><name sortKey="Schulman, H" sort="Schulman, H" uniqKey="Schulman H" first="H" last="Schulman">H. Schulman</name><name sortKey="Stryer, L" sort="Stryer, L" uniqKey="Stryer L" first="L" last="Stryer">L. Stryer</name></noCountry><country name="États-Unis"><region name="Californie"><name sortKey="Wagner, J A" sort="Wagner, J A" uniqKey="Wagner J" first="J A" last="Wagner">J A Wagner</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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