Phosphorylation Decreases Ubiquitylation of the Thiazide-sensitive Cotransporter NCC and Subsequent Clathrin-mediated Endocytosis

Rosenbæk, Lena Lindtoft; Kortenoeven, Marleen L. A.; Aroankins, Takwa S.; Fenton, Robert A.

doi:10.1074/jbc.m113.543710

Cited by 66 publications

(73 citation statements)

References 53 publications

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“…This adds to the evidence that SPAK-mediated phosphorylation acts primarily to increase activity of individual cotransporters without affecting the amount of NCC on the surface (16,19,21). However, there is also evidence in the literature that phosphorylation can affect surface expression (12,24). A recent study demonstrated that phospho-mimicking NCC mutants (replacing T53, T8, and S71 with aspartic acids) have reduced endocytosis resulting in greater surface expression compared with wild-type NCC.…”

Section: Discussionmentioning

confidence: 85%

Mechanisms of angiotensin II stimulation of NCC are time-dependent in mDCT15 cells

Mistry

Hanson

et al. 2015

American Journal of Physiology-Renal Physiology

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-Angiotensin II (ANG II) increases thiazide-sensitive sodium-chloride cotransporter (NCC) activity both acutely and chronically. ANG II has been implicated as a switch that turns WNK4 from an inhibitor of NCC into an activator of NCC, and ANG II's effect on NCC appears to require WNK4. Chronically, ANG II stimulation of NCC results in an increase in total and phosphorylated NCC, but the role of NCC phosphorylation in acute ANG II actions is unclear. Here, using a mammalian cell model with robust native NCC activity, we corroborate the role that ANG II plays in WNK4 regulation and clarify the role of Ste20-related proline alanine-rich kinase (SPAK)-induced NCC phosphorylation in ANG II action. ANG II was noted to have a biphasic effect on NCC, with a peak increase in NCC activity in the physiologic range of 10 Ϫ11 M ANG II. This effect was apparent as early as 15 min and remained sustained through 120 min. These changes correlated with significant increases in NCC surface protein expression. Knockdown of WNK4 expression sharply attenuated the effect of ANG II. SPAK knockdown did not affect ANG II action at early time points (15 and 30 min), but it did attenuate the response at 60 min. Correspondingly, NCC phosphorylation did not increase at 15 or 30 min, but increased significantly at 60 min. We therefore conclude that within minutes of an increase in ANG II, NCC is rapidly trafficked to the cell surface in a phosphorylation-independent but WNK4-dependent manner. Then, after 60 min, ANG II induces SPAK-dependent phosphorylation of NCC.thiazide-sensitive sodium-chloride cotransporter; distal convoluted tubule; WNK4 expression THE THIAZIDE-SENSITIVE sodium-chloride cotransporter (NCC) is the salt-reabsorptive pathway localized to the apical membrane of the mammalian distal convoluted tubule (DCT) that is responsible for reabsorbing 5-10% of the filtered load of sodium (9). Now, almost 60 years after the introduction of the first thiazide diuretic (8), pharmacological inhibition of NCC by thiazide diuretics is recommended as first line treatment for essential hypertension (6). NCC has also been shown to play a role in genetic disorders of hypotension and hypertension (7,18,27,33,34).Over the last decade it has become clear that the reninangiotensin-aldosterone system (RAAS) is the primary physiological regulator of NCC. First reported were the actions of aldosterone on NCC. Chronic aldosterone exposure (3-8 days) increases total and phosphorylated NCC and results in an increase in thiazide-sensitive sodium reabsorption (14,20,32).Acute aldosterone (12-36 h) stimulation, by contrast, increases NCC activity and phosphorylation without a change in total NCC (16). More recently, the effects of ANG II on the cotransporter have been described. ANG II has been implicated as a switch that turns WNK4 [With-No-Lysine (K) 4] from an inhibitor of NCC into an activator of NCC (4, 5, 25). Additionally, ANG II's effect on NCC appears to require WNK4. This dependence of ANG II's effects on WNK4 has been investigated chronically wit...

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Section: Discussionmentioning

confidence: 85%

Mechanisms of angiotensin II stimulation of NCC are time-dependent in mDCT15 cells

Mistry

Hanson

et al. 2015

American Journal of Physiology-Renal Physiology

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“…It has been reported that the SPAK-induced phosphorylation of NCC inhibited ubiquitylation of the thiazide-sensitive NCC in the DCT. 41 Although the disruption of cav-1 reduced the expression of NCC, the net renal Na + excretion was not significantly larger in cav-1 KO mice than in WT mice. It is possible that a decrease in NCC expression in the DCT should increase Na + delivery to the CNT/CCD, thereby stimulating Na + absorption through ENaC in these nephron segments, thereby offsetting the effect of low NCC expression.…”

Section: +mentioning

confidence: 99%

Caveolin-1 Deficiency Inhibits the Basolateral K+ Channels in the Distal Convoluted Tubule and Impairs Renal K+ and Mg2+ Transport

Wang

Zhang

et al. 2015

Journal of the American Society of Nephrology

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Kir4.1/Kir5.1, in the basolateral membrane of the early DCT (DCT1) in both wild-type (WT) and cav-1-knockout (KO) mice. However, the activity of this basolateral 40-pS K + channel was lower in KO mice than in WT mice. Moreover, the K + reversal potential (an indication of membrane potential) was less negative in the DCT1 of KO mice than in the DCT1 of WT mice. Western blot analysis demonstrated that cav-1 deficiency decreased the expression of the Na + /Cl -cotransporter and Ste20-proline-alanine-rich kinase (SPAK) but increased the expression of epithelial Na + channel-a. Furthermore, the urinary excretion of Mg 2+ and K + was significantly higher in KO mice than in WT mice, and KO mice developed hypomagnesemia, hypocalcemia, and hypokalemia. We conclude that disruption of cav-1 decreases basolateral K + channel activity and depolarizes the cell membrane potential in the DCT1 at least in part by suppressing the stimulatory effect of c-Src on Kcnj10. Furthermore, the decrease in Kcnj10 and Na + /Cl -cotransporter expression induced by cav-1 deficiency may underlie the compromised renal transport of Mg 2+ , Ca 2+ , and K + .

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“…Evidence suggests that NKCC2 phosphorylation at the serine/threonine residues in the amino-terminal domain (Thr-95, Thr-100, and Thr-105 of human NKCC2), which are conserved in NCC and NKCC1, activate the cotransporter (46, 47, 106); however, it is unclear whether phosphorylation of these residues is associated with activation of the cotransporter that is already in the plasma membrane and/or upregulation of the NKCC2 in the apical membrane. In this regard, data from NCC studies strongly suggest that SPAK/ OSR1 phosphorylates the cotransporter that is already in the plasma membrane, and the phosphorylated transporter becomes resistant to ubiquitylation and clathrin-related endocytosis, ultimately increasing the NCC expression in the plasma membrane (99,118). Given the similarities between NCC and NKCC2, it is possible that a similar mechanism applies for NKCC2.…”

Section: F135 Slc12 Cotransporters In the Kidneymentioning

confidence: 99%

“…The K ϩ modulation of NCC activity has gained much attention (28, 100) because it raised questions about the previously accepted role of aldosterone in stimulating NCC expression/activity (6, 61). It is known that NCC can be regulated by ubiquitylation (63,118), which can occur via the HECT-E3 ligase Nedd4-2 (5) or through modulation of WNK1 ubiquitylation (119). However, aldosterone administration increases urinary K ϩ loss, which, in turn, will reduce serum K ϩ levels and, thus, stimulate NCC activity.…”

Section: F137 Slc12 Cotransporters In the Kidneymentioning

confidence: 99%

Physiological role of SLC12 family members in the kidney

Bazúa‐Valenti

Castañeda‐Bueno

Gamba

2016

American Journal of Physiology-Renal Physiology

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The solute carrier family 12, as numbered according to Human Genome Organisation (HUGO) nomenclature, encodes the electroneutral cation-coupled chloride cotransporters that are expressed in many cells and tissues; they play key roles in important physiological events, such as cell volume regulation, modulation of the intracellular chloride concentration, and transepithelial ion transport. Most of these family members are expressed in specific regions of the nephron. The Na-K-2Cl cotransporter NKCC2, which is located in the thick ascending limb, and the Na-Cl cotransporter, which is located in the distal convoluted tubule, play important roles in salt reabsorption and serve as the receptors for loop and thiazide diuretics, respectively (Thiazide diuretics are among the most commonly prescribed drugs in the world.). The activity of these transporters correlates with blood pressure levels; thus, their regulation has been a subject of intense research for more than a decade. The K-Cl cotransporters KCC1, KCC3, and KCC4 are expressed in several nephron segments, and their role in renal physiology is less understood but nevertheless important. Evidence suggests that they are involved in modulating proximal tubule glucose reabsorption, thick ascending limb salt reabsorption and collecting duct proton secretion. In this work, we present an overview of the physiological roles of these transporters in the kidney, with particular emphasis on the knowledge gained in the past few years.

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Phosphorylation Decreases Ubiquitylation of the Thiazide-sensitive Cotransporter NCC and Subsequent Clathrin-mediated Endocytosis

Cited by 66 publications

References 53 publications

Mechanisms of angiotensin II stimulation of NCC are time-dependent in mDCT15 cells

Mechanisms of angiotensin II stimulation of NCC are time-dependent in mDCT15 cells

Caveolin-1 Deficiency Inhibits the Basolateral K+ Channels in the Distal Convoluted Tubule and Impairs Renal K+ and Mg2+ Transport

Physiological role of SLC12 family members in the kidney

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