Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter

Aljona Borschewski, Nina Himmerkus, Christin Boldt, Katharina I. Blankenstein, James (Jim) McCormick, Rebecca Lazelle, Thomas E. Willnow, Vera Jankowski, Allein Plain, Markus Bleich, David Ellison, Sebastian Bachmann, Kerim Mutig

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The furosemide-sensitive Na+-K+-2Cl--cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick ascending limb (TAL) and drives the urine concentrating mechanism. NKCC2 activity ismodulated by N-terminal phosphorylation and dephosphorylation. Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phosphatases that deactivate NKCC2. Inhibition of calcineurin phosphatase has been shown to stimulate transport in the TAL and the distal convoluted tubule. Here, we identified NKCC2 as a target of the calcineurin Aβ isoform. Short-term cyclosporine administration inmice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine increased the chloride affinity and transport activity of NKCC2. Because sorting-related receptor with Atype repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to test whether SORLA is involved in calcineurin-dependent modulation of NKCC2. SORLA-deficient mice showed more calcineurin Aβ in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with littermate controls. In contrast, overexpression of SORLA in cultured cells reduced the abundance of endogenous calcineurin Aβ. Cyclosporine administration rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficientmice, and a functional interaction between calcineurin Aβ and SORLAwas further corroborated by binding assays in rat kidney extracts. In summary, we have shown that calcineurin Aβ and SORLA are key components in the phosphoregulation of NKCC2. These results may have clinical implications for immunosuppressive therapy using calcineurin inhibitors.

Original languageEnglish (US)
Pages (from-to)107-119
Number of pages13
JournalJournal of the American Society of Nephrology
Volume27
Issue number1
DOIs
StatePublished - Jan 1 2016

Fingerprint

Calcineurin
Kidney
Extremities
Cyclosporine
Member 1 Solute Carrier Family 12
Phosphorylation
Protein-Serine-Threonine Kinases
Furosemide
Immunosuppressive Agents
Phosphoric Monoester Hydrolases
Knockout Mice
Chlorides
Cultured Cells
Protein Isoforms
Urine

ASJC Scopus subject areas

  • Nephrology

Cite this

Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter. / Borschewski, Aljona; Himmerkus, Nina; Boldt, Christin; Blankenstein, Katharina I.; McCormick, James (Jim); Lazelle, Rebecca; Willnow, Thomas E.; Jankowski, Vera; Plain, Allein; Bleich, Markus; Ellison, David; Bachmann, Sebastian; Mutig, Kerim.

In: Journal of the American Society of Nephrology, Vol. 27, No. 1, 01.01.2016, p. 107-119.

Research output: Contribution to journalArticle

Borschewski, A, Himmerkus, N, Boldt, C, Blankenstein, KI, McCormick, JJ, Lazelle, R, Willnow, TE, Jankowski, V, Plain, A, Bleich, M, Ellison, D, Bachmann, S & Mutig, K 2016, 'Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter', Journal of the American Society of Nephrology, vol. 27, no. 1, pp. 107-119. https://doi.org/10.1681/ASN.2014070728
Borschewski, Aljona ; Himmerkus, Nina ; Boldt, Christin ; Blankenstein, Katharina I. ; McCormick, James (Jim) ; Lazelle, Rebecca ; Willnow, Thomas E. ; Jankowski, Vera ; Plain, Allein ; Bleich, Markus ; Ellison, David ; Bachmann, Sebastian ; Mutig, Kerim. / Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter. In: Journal of the American Society of Nephrology. 2016 ; Vol. 27, No. 1. pp. 107-119.
@article{80fb745fd9ac4b889e40889fe02f964a,
title = "Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter",
abstract = "The furosemide-sensitive Na+-K+-2Cl--cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick ascending limb (TAL) and drives the urine concentrating mechanism. NKCC2 activity ismodulated by N-terminal phosphorylation and dephosphorylation. Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phosphatases that deactivate NKCC2. Inhibition of calcineurin phosphatase has been shown to stimulate transport in the TAL and the distal convoluted tubule. Here, we identified NKCC2 as a target of the calcineurin Aβ isoform. Short-term cyclosporine administration inmice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine increased the chloride affinity and transport activity of NKCC2. Because sorting-related receptor with Atype repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to test whether SORLA is involved in calcineurin-dependent modulation of NKCC2. SORLA-deficient mice showed more calcineurin Aβ in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with littermate controls. In contrast, overexpression of SORLA in cultured cells reduced the abundance of endogenous calcineurin Aβ. Cyclosporine administration rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficientmice, and a functional interaction between calcineurin Aβ and SORLAwas further corroborated by binding assays in rat kidney extracts. In summary, we have shown that calcineurin Aβ and SORLA are key components in the phosphoregulation of NKCC2. These results may have clinical implications for immunosuppressive therapy using calcineurin inhibitors.",
author = "Aljona Borschewski and Nina Himmerkus and Christin Boldt and Blankenstein, {Katharina I.} and McCormick, {James (Jim)} and Rebecca Lazelle and Willnow, {Thomas E.} and Vera Jankowski and Allein Plain and Markus Bleich and David Ellison and Sebastian Bachmann and Kerim Mutig",
year = "2016",
month = "1",
day = "1",
doi = "10.1681/ASN.2014070728",
language = "English (US)",
volume = "27",
pages = "107--119",
journal = "Journal of the American Society of Nephrology : JASN",
issn = "1046-6673",
publisher = "American Society of Nephrology",
number = "1",

}

TY - JOUR

T1 - Calcineurin and Sorting-Related Receptor with A-Type Repeats Interact to Regulate the Renal Na+-K+-2Cl- Cotransporter

AU - Borschewski, Aljona

AU - Himmerkus, Nina

AU - Boldt, Christin

AU - Blankenstein, Katharina I.

AU - McCormick, James (Jim)

AU - Lazelle, Rebecca

AU - Willnow, Thomas E.

AU - Jankowski, Vera

AU - Plain, Allein

AU - Bleich, Markus

AU - Ellison, David

AU - Bachmann, Sebastian

AU - Mutig, Kerim

PY - 2016/1/1

Y1 - 2016/1/1

N2 - The furosemide-sensitive Na+-K+-2Cl--cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick ascending limb (TAL) and drives the urine concentrating mechanism. NKCC2 activity ismodulated by N-terminal phosphorylation and dephosphorylation. Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phosphatases that deactivate NKCC2. Inhibition of calcineurin phosphatase has been shown to stimulate transport in the TAL and the distal convoluted tubule. Here, we identified NKCC2 as a target of the calcineurin Aβ isoform. Short-term cyclosporine administration inmice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine increased the chloride affinity and transport activity of NKCC2. Because sorting-related receptor with Atype repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to test whether SORLA is involved in calcineurin-dependent modulation of NKCC2. SORLA-deficient mice showed more calcineurin Aβ in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with littermate controls. In contrast, overexpression of SORLA in cultured cells reduced the abundance of endogenous calcineurin Aβ. Cyclosporine administration rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficientmice, and a functional interaction between calcineurin Aβ and SORLAwas further corroborated by binding assays in rat kidney extracts. In summary, we have shown that calcineurin Aβ and SORLA are key components in the phosphoregulation of NKCC2. These results may have clinical implications for immunosuppressive therapy using calcineurin inhibitors.

AB - The furosemide-sensitive Na+-K+-2Cl--cotransporter (NKCC2) is crucial for NaCl reabsorption in kidney thick ascending limb (TAL) and drives the urine concentrating mechanism. NKCC2 activity ismodulated by N-terminal phosphorylation and dephosphorylation. Serine-threonine kinases that activate NKCC2 have been identified, but less is known about phosphatases that deactivate NKCC2. Inhibition of calcineurin phosphatase has been shown to stimulate transport in the TAL and the distal convoluted tubule. Here, we identified NKCC2 as a target of the calcineurin Aβ isoform. Short-term cyclosporine administration inmice augmented the abundance of phospho-NKCC2, and treatment of isolated TAL with cyclosporine increased the chloride affinity and transport activity of NKCC2. Because sorting-related receptor with Atype repeats (SORLA) may affect NKCC2 phosphoregulation, we used SORLA-knockout mice to test whether SORLA is involved in calcineurin-dependent modulation of NKCC2. SORLA-deficient mice showed more calcineurin Aβ in the apical region of TAL cells and less NKCC2 phosphorylation and activity compared with littermate controls. In contrast, overexpression of SORLA in cultured cells reduced the abundance of endogenous calcineurin Aβ. Cyclosporine administration rapidly normalized the abundance of phospho-NKCC2 in SORLA-deficientmice, and a functional interaction between calcineurin Aβ and SORLAwas further corroborated by binding assays in rat kidney extracts. In summary, we have shown that calcineurin Aβ and SORLA are key components in the phosphoregulation of NKCC2. These results may have clinical implications for immunosuppressive therapy using calcineurin inhibitors.

UR - http://www.scopus.com/inward/record.url?scp=84954447669&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84954447669&partnerID=8YFLogxK

U2 - 10.1681/ASN.2014070728

DO - 10.1681/ASN.2014070728

M3 - Article

C2 - 25967121

AN - SCOPUS:84954447669

VL - 27

SP - 107

EP - 119

JO - Journal of the American Society of Nephrology : JASN

JF - Journal of the American Society of Nephrology : JASN

SN - 1046-6673

IS - 1

ER -