Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel

Abstract Activity of the Epithelial Na+ Channel (ENaC) in the distal nephron fine-tunes renal sodium excretion. Appropriate sodium excretion is a key factor in the regulation of blood pressure. Consequently, abnormalities in ENaC function can cause hypertension. Casein Kinase II (CKII) phosphorylate...

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Autores principales:	Tarek Mohamed Abd El-Aziz, Antonio G. Soares, Elena Mironova, Nina Boiko, Amanpreet Kaur, Crystal R. Archer, James D. Stockand, Jonathan M. Berman
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dab1b85344ad40b297a5b5d8718ba07c
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spelling	oai:doaj.org-article:dab1b85344ad40b297a5b5d8718ba07c2021-12-02T16:08:07ZMechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel10.1038/s41598-021-94118-32045-2322https://doaj.org/article/dab1b85344ad40b297a5b5d8718ba07c2021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94118-3https://doaj.org/toc/2045-2322Abstract Activity of the Epithelial Na+ Channel (ENaC) in the distal nephron fine-tunes renal sodium excretion. Appropriate sodium excretion is a key factor in the regulation of blood pressure. Consequently, abnormalities in ENaC function can cause hypertension. Casein Kinase II (CKII) phosphorylates ENaC. The CKII phosphorylation site in ENaC resides within a canonical “anchor” ankyrin binding motif. CKII-dependent phosphorylation of ENaC is necessary and sufficient to increase channel activity and is thought to influence channel trafficking in a manner that increases activity. We test here the hypothesis that phosphorylation of ENaC by CKII within an anchor motif is necessary for ankyrin-3 (Ank-3) regulation of the channel, which is required for normal channel locale and function, and the proper regulation of renal sodium excretion. This was addressed using a fluorescence imaging strategy combining total internal reflection fluorescence (TIRF) microscopy with fluorescence recovery after photobleaching (FRAP) to quantify ENaC expression in the plasma membrane in living cells; and electrophysiology to quantify ENaC activity in split-open collecting ducts from principal cell-specific Ank-3 knockout mice. Sodium excretion studies also were performed in parallel in this knockout mouse. In addition, we substituted a key serine residue in the consensus CKII site in β-ENaC with alanine to abrogate phosphorylation and disrupt the anchor motif. Findings show that disrupting CKII signaling decreases ENaC activity by decreasing expression in the plasma membrane. In the principal cell-specific Ank-3 KO mouse, ENaC activity and sodium excretion were significantly decreased and increased, respectively. These results are consistent with CKII phosphorylation of ENaC functioning as a “switch” that favors Ank-3 binding to increase channel activity.Tarek Mohamed Abd El-AzizAntonio G. SoaresElena MironovaNina BoikoAmanpreet KaurCrystal R. ArcherJames D. StockandJonathan M. BermanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tarek Mohamed Abd El-Aziz Antonio G. Soares Elena Mironova Nina Boiko Amanpreet Kaur Crystal R. Archer James D. Stockand Jonathan M. Berman Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
description	Abstract Activity of the Epithelial Na+ Channel (ENaC) in the distal nephron fine-tunes renal sodium excretion. Appropriate sodium excretion is a key factor in the regulation of blood pressure. Consequently, abnormalities in ENaC function can cause hypertension. Casein Kinase II (CKII) phosphorylates ENaC. The CKII phosphorylation site in ENaC resides within a canonical “anchor” ankyrin binding motif. CKII-dependent phosphorylation of ENaC is necessary and sufficient to increase channel activity and is thought to influence channel trafficking in a manner that increases activity. We test here the hypothesis that phosphorylation of ENaC by CKII within an anchor motif is necessary for ankyrin-3 (Ank-3) regulation of the channel, which is required for normal channel locale and function, and the proper regulation of renal sodium excretion. This was addressed using a fluorescence imaging strategy combining total internal reflection fluorescence (TIRF) microscopy with fluorescence recovery after photobleaching (FRAP) to quantify ENaC expression in the plasma membrane in living cells; and electrophysiology to quantify ENaC activity in split-open collecting ducts from principal cell-specific Ank-3 knockout mice. Sodium excretion studies also were performed in parallel in this knockout mouse. In addition, we substituted a key serine residue in the consensus CKII site in β-ENaC with alanine to abrogate phosphorylation and disrupt the anchor motif. Findings show that disrupting CKII signaling decreases ENaC activity by decreasing expression in the plasma membrane. In the principal cell-specific Ank-3 KO mouse, ENaC activity and sodium excretion were significantly decreased and increased, respectively. These results are consistent with CKII phosphorylation of ENaC functioning as a “switch” that favors Ank-3 binding to increase channel activity.
format	article
author	Tarek Mohamed Abd El-Aziz Antonio G. Soares Elena Mironova Nina Boiko Amanpreet Kaur Crystal R. Archer James D. Stockand Jonathan M. Berman
author_facet	Tarek Mohamed Abd El-Aziz Antonio G. Soares Elena Mironova Nina Boiko Amanpreet Kaur Crystal R. Archer James D. Stockand Jonathan M. Berman
author_sort	Tarek Mohamed Abd El-Aziz
title	Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
title_short	Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
title_full	Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
title_fullStr	Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
title_full_unstemmed	Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel
title_sort	mechanisms and consequences of casein kinase ii and ankyrin-3 regulation of the epithelial na+ channel
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/dab1b85344ad40b297a5b5d8718ba07c
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Mechanisms and consequences of casein kinase II and ankyrin-3 regulation of the epithelial Na+ channel

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