Anterograde Trafficking of KCa3.1 in Polarized Epithelia Is Rab1- and Rab8-Dependent and Recycling Endosome-Independent
Identifieur interne : 001598 ( Ncbi/Curation ); précédent : 001597; suivant : 001599Anterograde Trafficking of KCa3.1 in Polarized Epithelia Is Rab1- and Rab8-Dependent and Recycling Endosome-Independent
Auteurs : Claudia A. Bertuccio [États-Unis] ; Shih-Liang Lee [Nouvelle-Zélande] ; Guangyu Wu [États-Unis] ; Michael B. Butterworth [États-Unis] ; Kirk L. Hamilton [Nouvelle-Zélande] ; Daniel C. Devor [États-Unis]Source :
- PLoS ONE [ 1932-6203 ] ; 2014.
Descripteurs français
- KwdFr :
- Animaux, Appareil de Golgi (métabolisme), Canaux potassiques calcium-dépendants de conductance intermédiaire (métabolisme), Cellules épithéliales (cytologie), Cellules épithéliales (métabolisme), Chiens, Endosomes (métabolisme), Humains, Lignée cellulaire, Membrane cellulaire (métabolisme), Polarité de la cellule, Protéines G rab (métabolisme), Protéines G rab1 (métabolisme), Réticulum endoplasmique (métabolisme), Transport de protéines, Ubiquitinylation.
- MESH :
- cytologie : Cellules épithéliales.
- métabolisme : Appareil de Golgi, Canaux potassiques calcium-dépendants de conductance intermédiaire, Cellules épithéliales, Endosomes, Membrane cellulaire, Protéines G rab, Protéines G rab1, Réticulum endoplasmique.
- Animaux, Chiens, Humains, Lignée cellulaire, Polarité de la cellule, Transport de protéines, Ubiquitinylation.
English descriptors
- KwdEn :
- Animals, Cell Line, Cell Membrane (metabolism), Cell Polarity, Dogs, Endoplasmic Reticulum (metabolism), Endosomes (metabolism), Epithelial Cells (cytology), Epithelial Cells (metabolism), Golgi Apparatus (metabolism), Humans, Intermediate-Conductance Calcium-Activated Potassium Channels (metabolism), Protein Transport, Ubiquitination, rab GTP-Binding Proteins (metabolism), rab1 GTP-Binding Proteins (metabolism).
- MESH :
- chemical , metabolism : Intermediate-Conductance Calcium-Activated Potassium Channels, rab GTP-Binding Proteins, rab1 GTP-Binding Proteins.
- cytology : Epithelial Cells.
- metabolism : Cell Membrane, Endoplasmic Reticulum, Endosomes, Epithelial Cells, Golgi Apparatus.
- Animals, Cell Line, Cell Polarity, Dogs, Humans, Protein Transport, Ubiquitination.
Abstract
The intermediate conductance, Ca2+-activated K+ channel (KCa3.1) targets to the basolateral (BL) membrane in polarized epithelia where it plays a key role in transepithelial ion transport. However, there are no studies defining the anterograde and retrograde trafficking of KCa3.1 in polarized epithelia. Herein, we utilize
Url:
DOI: 10.1371/journal.pone.0092013
PubMed: 24632741
PubMed Central: 3954861
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PMC:3954861Le document en format XML
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Cell Membrane (metabolism)</term>
<term>Cell Polarity</term>
<term>Dogs</term>
<term>Endoplasmic Reticulum (metabolism)</term>
<term>Endosomes (metabolism)</term>
<term>Epithelial Cells (cytology)</term>
<term>Epithelial Cells (metabolism)</term>
<term>Golgi Apparatus (metabolism)</term>
<term>Humans</term>
<term>Intermediate-Conductance Calcium-Activated Potassium Channels (metabolism)</term>
<term>Protein Transport</term>
<term>Ubiquitination</term>
<term>rab GTP-Binding Proteins (metabolism)</term>
<term>rab1 GTP-Binding Proteins (metabolism)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Appareil de Golgi (métabolisme)</term>
<term>Canaux potassiques calcium-dépendants de conductance intermédiaire (métabolisme)</term>
<term>Cellules épithéliales (cytologie)</term>
<term>Cellules épithéliales (métabolisme)</term>
<term>Chiens</term>
<term>Endosomes (métabolisme)</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Membrane cellulaire (métabolisme)</term>
<term>Polarité de la cellule</term>
<term>Protéines G rab (métabolisme)</term>
<term>Protéines G rab1 (métabolisme)</term>
<term>Réticulum endoplasmique (métabolisme)</term>
<term>Transport de protéines</term>
<term>Ubiquitinylation</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Intermediate-Conductance Calcium-Activated Potassium Channels</term>
<term>rab GTP-Binding Proteins</term>
<term>rab1 GTP-Binding Proteins</term>
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<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Epithelial Cells</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Cell Membrane</term>
<term>Endoplasmic Reticulum</term>
<term>Endosomes</term>
<term>Epithelial Cells</term>
<term>Golgi Apparatus</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Appareil de Golgi</term>
<term>Canaux potassiques calcium-dépendants de conductance intermédiaire</term>
<term>Cellules épithéliales</term>
<term>Endosomes</term>
<term>Membrane cellulaire</term>
<term>Protéines G rab</term>
<term>Protéines G rab1</term>
<term>Réticulum endoplasmique</term>
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<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Cell Polarity</term>
<term>Dogs</term>
<term>Humans</term>
<term>Protein Transport</term>
<term>Ubiquitination</term>
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<term>Chiens</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Polarité de la cellule</term>
<term>Transport de protéines</term>
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<front><div type="abstract" xml:lang="en"><p>The intermediate conductance, Ca<sup>2+</sup>
-activated K<sup>+</sup>
channel (KCa3.1) targets to the basolateral (BL) membrane in polarized epithelia where it plays a key role in transepithelial ion transport. However, there are no studies defining the anterograde and retrograde trafficking of KCa3.1 in polarized epithelia. Herein, we utilize <underline>B</underline>
iotin <underline>L</underline>
igase <underline>A</underline>
cceptor <underline>P</underline>
eptide (BLAP)-tagged KCa3.1 to address these trafficking steps in polarized epithelia, using MDCK, Caco-2 and FRT cells. We demonstrate that KCa3.1 is exclusively targeted to the BL membrane in these cells when grown on filter supports. Following endocytosis, KCa3.1 degradation is prevented by inhibition of lysosomal/proteosomal pathways. Further, the ubiquitylation of KCa3.1 is increased following endocytosis from the BL membrane and PR-619, a deubiquitylase inhibitor, prevents degradation, indicating KCa3.1 is targeted for degradation by ubiquitylation. We demonstrate that KCa3.1 is targeted to the BL membrane in polarized LLC-PK<sub>1</sub>
cells which lack the μ1B subunit of the AP-1 complex, indicating BL targeting of KCa3.1 is independent of μ1B. As Rabs 1, 2, 6 and 8 play roles in ER/Golgi exit and trafficking of proteins to the BL membrane, we evaluated the role of these Rabs in the trafficking of KCa3.1. In the presence of dominant negative Rab1 or Rab8, KCa3.1 cell surface expression was significantly reduced, whereas Rabs 2 and 6 had no effect. We also co-immunoprecipitated KCa3.1 with both Rab1 and Rab8. These results suggest these Rabs are necessary for the anterograde trafficking of KCa3.1. Finally, we determined whether KCa3.1 traffics directly to the BL membrane or through recycling endosomes in MDCK cells. For these studies, we used either recycling endosome ablation or dominant negative RME-1 constructs and determined that KCa3.1 is trafficked directly to the BL membrane rather than via recycling endosomes. These results are the first to describe the anterograde and retrograde trafficking of KCa3.1 in polarized epithelia cells.</p>
</div>
</front>
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