Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3

James (Jim) McCormick, Chao-Ling Yang, Chong Zhang, Brittney Davidge, Katharina I. Blankenstein, Andrew S. Terker, Bethzaida Yarbrough, Nicholas P. Meermeier, Hae J. Park, Belinda McCully, Mark West, Aljona Borschewski, Nina Himmerkus, Markus Bleich, Sebastian Bachmann, Kerim Mutig, Eduardo R. Argaiz, Gerardo Gamba, Jeffrey D. Singer, David Ellison

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Abstract

Familial hyperkalemic hypertension (FHHt) is a monogenic disease resulting from mutations in genes encoding WNK kinases, the ubiquitin scaffold protein cullin 3 (CUL3), or the substrate adaptor kelch-like 3 (KLHL3). Disease-associated CUL3 mutations abrogate WNK kinase degradation in cells, but it is not clear how mutant forms of CUL3 promote WNK stability. Here, we demonstrated that an FHHt-causing CUL3 mutant (CUL3 Δ403-459) not only retains the ability to bind and ubiquitylate WNK kinases and KLHL3 in cells, but is also more heavily neddylated and activated than WT CUL3. In cells, activated CUL3 Δ403-459 depleted KLHL3, preventing WNK degradation, despite increased CUL3-mediated WNK ubiquitylation; therefore, CUL3 loss in kidney should phenocopy FHHt in murine models. As predicted, nephron-specific deletion of Cul3 in mice did increase WNK kinase levels and the abundance of phosphorylated Na-Cl cotransporter (NCC). Over time, however, Cul3 deletion caused renal dysfunction, including hypochloremic alkalosis, diabetes insipidus, and salt-sensitive hypotension, with depletion of sodium potassium chloride cotransporter 2 and aquaporin 2. Moreover, these animals exhibited renal inflammation, fibrosis, and increased cyclin E. These results indicate that FHHt-associated CUL3 Δ403-459 targets KLHL3 for degradation, thereby preventing WNK degradation, whereas general loss of CUL3 activity - while also impairing WNK degradation - has widespread toxic effects in the kidney.

Original languageEnglish (US)
Pages (from-to)4723-4736
Number of pages14
JournalJournal of Clinical Investigation
Volume124
Issue number11
DOIs
StatePublished - Nov 3 2014

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Cullin Proteins
Hypertension
Phosphotransferases
Kidney
Member 3 Solute Carrier Family 12
Sodium-Potassium-Chloride Symporters
Aquaporin 2
Cyclin E
Alkalosis
Diabetes Insipidus
Mutation
Poisons
Ubiquitination
Nephrons
Ubiquitin
Hypotension

ASJC Scopus subject areas

  • Medicine(all)

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Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3. / McCormick, James (Jim); Yang, Chao-Ling; Zhang, Chong; Davidge, Brittney; Blankenstein, Katharina I.; Terker, Andrew S.; Yarbrough, Bethzaida; Meermeier, Nicholas P.; Park, Hae J.; McCully, Belinda; West, Mark; Borschewski, Aljona; Himmerkus, Nina; Bleich, Markus; Bachmann, Sebastian; Mutig, Kerim; Argaiz, Eduardo R.; Gamba, Gerardo; Singer, Jeffrey D.; Ellison, David.

In: Journal of Clinical Investigation, Vol. 124, No. 11, 03.11.2014, p. 4723-4736.

Research output: Contribution to journalArticle

McCormick, JJ, Yang, C-L, Zhang, C, Davidge, B, Blankenstein, KI, Terker, AS, Yarbrough, B, Meermeier, NP, Park, HJ, McCully, B, West, M, Borschewski, A, Himmerkus, N, Bleich, M, Bachmann, S, Mutig, K, Argaiz, ER, Gamba, G, Singer, JD & Ellison, D 2014, 'Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3', Journal of Clinical Investigation, vol. 124, no. 11, pp. 4723-4736. https://doi.org/10.1172/JCI76126
McCormick, James (Jim) ; Yang, Chao-Ling ; Zhang, Chong ; Davidge, Brittney ; Blankenstein, Katharina I. ; Terker, Andrew S. ; Yarbrough, Bethzaida ; Meermeier, Nicholas P. ; Park, Hae J. ; McCully, Belinda ; West, Mark ; Borschewski, Aljona ; Himmerkus, Nina ; Bleich, Markus ; Bachmann, Sebastian ; Mutig, Kerim ; Argaiz, Eduardo R. ; Gamba, Gerardo ; Singer, Jeffrey D. ; Ellison, David. / Hyperkalemic hypertension-associated cullin 3 promotes WNK signaling by degrading KLHL3. In: Journal of Clinical Investigation. 2014 ; Vol. 124, No. 11. pp. 4723-4736.
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abstract = "Familial hyperkalemic hypertension (FHHt) is a monogenic disease resulting from mutations in genes encoding WNK kinases, the ubiquitin scaffold protein cullin 3 (CUL3), or the substrate adaptor kelch-like 3 (KLHL3). Disease-associated CUL3 mutations abrogate WNK kinase degradation in cells, but it is not clear how mutant forms of CUL3 promote WNK stability. Here, we demonstrated that an FHHt-causing CUL3 mutant (CUL3 Δ403-459) not only retains the ability to bind and ubiquitylate WNK kinases and KLHL3 in cells, but is also more heavily neddylated and activated than WT CUL3. In cells, activated CUL3 Δ403-459 depleted KLHL3, preventing WNK degradation, despite increased CUL3-mediated WNK ubiquitylation; therefore, CUL3 loss in kidney should phenocopy FHHt in murine models. As predicted, nephron-specific deletion of Cul3 in mice did increase WNK kinase levels and the abundance of phosphorylated Na-Cl cotransporter (NCC). Over time, however, Cul3 deletion caused renal dysfunction, including hypochloremic alkalosis, diabetes insipidus, and salt-sensitive hypotension, with depletion of sodium potassium chloride cotransporter 2 and aquaporin 2. Moreover, these animals exhibited renal inflammation, fibrosis, and increased cyclin E. These results indicate that FHHt-associated CUL3 Δ403-459 targets KLHL3 for degradation, thereby preventing WNK degradation, whereas general loss of CUL3 activity - while also impairing WNK degradation - has widespread toxic effects in the kidney.",
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AU - McCormick, James (Jim)

AU - Yang, Chao-Ling

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AU - Davidge, Brittney

AU - Blankenstein, Katharina I.

AU - Terker, Andrew S.

AU - Yarbrough, Bethzaida

AU - Meermeier, Nicholas P.

AU - Park, Hae J.

AU - McCully, Belinda

AU - West, Mark

AU - Borschewski, Aljona

AU - Himmerkus, Nina

AU - Bleich, Markus

AU - Bachmann, Sebastian

AU - Mutig, Kerim

AU - Argaiz, Eduardo R.

AU - Gamba, Gerardo

AU - Singer, Jeffrey D.

AU - Ellison, David

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N2 - Familial hyperkalemic hypertension (FHHt) is a monogenic disease resulting from mutations in genes encoding WNK kinases, the ubiquitin scaffold protein cullin 3 (CUL3), or the substrate adaptor kelch-like 3 (KLHL3). Disease-associated CUL3 mutations abrogate WNK kinase degradation in cells, but it is not clear how mutant forms of CUL3 promote WNK stability. Here, we demonstrated that an FHHt-causing CUL3 mutant (CUL3 Δ403-459) not only retains the ability to bind and ubiquitylate WNK kinases and KLHL3 in cells, but is also more heavily neddylated and activated than WT CUL3. In cells, activated CUL3 Δ403-459 depleted KLHL3, preventing WNK degradation, despite increased CUL3-mediated WNK ubiquitylation; therefore, CUL3 loss in kidney should phenocopy FHHt in murine models. As predicted, nephron-specific deletion of Cul3 in mice did increase WNK kinase levels and the abundance of phosphorylated Na-Cl cotransporter (NCC). Over time, however, Cul3 deletion caused renal dysfunction, including hypochloremic alkalosis, diabetes insipidus, and salt-sensitive hypotension, with depletion of sodium potassium chloride cotransporter 2 and aquaporin 2. Moreover, these animals exhibited renal inflammation, fibrosis, and increased cyclin E. These results indicate that FHHt-associated CUL3 Δ403-459 targets KLHL3 for degradation, thereby preventing WNK degradation, whereas general loss of CUL3 activity - while also impairing WNK degradation - has widespread toxic effects in the kidney.

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