Phosphorylation of paracellin-1 at Ser217 by protein kinase A is essential for localization in tight junctions

Ikari, Akira; Matsumoto, Satomi; Hishida, Hitoshi; Takagi, Katsuyuki; Hayashi, Hideharu; Suzuki, Yuichi; Degawa, Masakuni; Miwa, Masao

doi:10.1242/jcs.02901

Cited by 95 publications

(96 citation statements)

References 57 publications

(59 reference statements)

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“…Aldosterone, the primary blood pressure-regulating hormone, upreguates paracellular Cl -permeability in a rat cortical collecting duct cell line RCCD2 and hyperphosphorylates CLDN4 (38). Although we have not studied the regulation of CLDN4 channel, claudin phosphorylation was demonstrated by Ikari et al (39) to play important roles in targeting claudin-16 to TJ. CLDN4 KO animal studies are currently under way to elucidate the in vivo role of CLDN4 in blood pressure control.…”

Section: Discussionmentioning

confidence: 79%

Claudin-4 forms paracellular chloride channel in the kidney and requires claudin-8 for tight junction localization

Hou

Renigunta²,

Yang³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

209

233

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Tight junctions (TJs) play a key role in mediating paracellular ion reabsorption in the kidney. The paracellular pathway in the collecting duct of the kidney is a predominant route for transepithelial chloride reabsorption that determines the extracellular NaCl content and the blood pressure. However, the molecular mechanisms underlying the paracellular chloride reabsorption in the collecting duct are not understood. Here we showed that in mouse kidney collecting duct cells, claudin-4 functioned as a Cl -channel. A positively charged lysine residue at position 65 of claudin-4 was critical for its anion selectivity. Claudin-4 was observed to interact with claudin-8 using several criteria. In the collecting duct cells, the assembly of claudin-4 into TJ strands required its interaction with claudin-8. Depletion of claudin-8 resulted in the loss of paracellular chloride conductance, through a mechanism involving its recruitment of claudin-4 during TJ assembly. Together, our data show that claudin-4 interacts with claudin-8 and that their association is required for the anion-selective paracellular pathway in the collecting duct, suggesting a mechanism for coupling chloride reabsorption with sodium reabsorption in the collecting duct.C hloride is the predominant extracellular ionic constituent and thereby determines extracellular fluid volume (ECFV) and blood pressure (1-3). Although only responsible for the reabsorption of 2-3% filtered chloride, the aldosterone-sensitive distal nephron (ASDN) plays a vital regulatory role in renal handling of salt, ECFV control, and managing blood pressure (4). The ASDN comprises the distal convoluted tubule (DCT), the connecting tubule (CNT), and the collecting duct. The collecting duct is characterized by a heterogeneous epithelium-the principal cells and intercalated cells (5). Sodium reabsorption in the collecting duct is an active process driven by the basolateral Na + /K + -ATPase; acts through the apical epithelial sodium channel (ENaC) in the principal cell (6); and is responsible for generating the lumen-negative transepithelial potential. This electrogenic transport step creates a favorable electrical driving force for luminal reabsorption of chloride and secretion of potassium and proton. Chloride is transported by two major mechanisms (1). Chloride is actively reabsorbed by an electroneutral Cl − /HCO − exchanger (Slc26a4: pendrin) localized to the apical membrane of the β-type intercalated cell (7). Choride exits this cell via a basolateral Cl − channel (2) and diffuses passively down electrochemical gradients via the paracellular channel in the tight junction (TJ).The TJ is the most apical member of the junctional complex found in vertebrate epithelia responsible for the barrier to movement of ions and molecules between apical and basal compartments, the paracellular pathway (8). TJs are composed of three transmembrane proteins: occludin, claudins, and junctional adhesion molecule (JAM). The claudins (CLDNs) are a 28-member family of tetraspan proteins that range in m...

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Section: Discussionmentioning

confidence: 79%

Claudin-4 forms paracellular chloride channel in the kidney and requires claudin-8 for tight junction localization

Hou

Renigunta²,

Yang³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

209

233

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“…The phosphorylation of claudin proteins is well-established and other claudins have been shown to be phosphorylated by various protein kinases. For example, claudin-1 is phosphorylated at Thr203 probably by MAPK [29], claudin-5 at Thr207 [32], claudin-16 at Ser217 [37], and claudin-4 by EphA2 at Tyr208 [35]. In addition claudin-1 and -4 are substrates for the WNK kinase, although no putative sites have been reported [33].…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, several studies have demonstrated the involvement of various kinases in the phosphorylation and regulation of claudin proteins [29][30][31][32][33][34][35][36][37], and we have recently shown that phosphorylation of claudin-3 by PKA can affect TJ properties in ovarian cancer cells [38]. Protein kinase C (PKC) isoforms are present in ovarian cancer and are known to modulate TJ function by phosphorylation of the proteins in the complex [24,34,[39][40][41][42][43], but it is unclear whether PKC can directly phosphorylate and regulate claudin proteins.…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of claudin-4 by PKCε regulates tight junction barrier function in ovarian cancer cells

D’Souza

Indig

Morin

2007

Experimental Cell Research

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Claudin proteins belong to a large family of transmembrane proteins essential to the formation and maintenance of tight junctions (TJs). In ovarian cancer, TJ protein claudin-4 is frequently overexpressed and may have roles in survival and invasion, but the molecular mechanisms underlying its regulation are poorly understood. In this report, we show that claudin-4 can be phosphorylated by protein kinase C (PKC) at Thr189 and Ser194 in ovarian cancer cells and overexpression of a claudin-4 mutant protein mimicking the phosphorylated state results in the disruption of the barrier function. Furthermore, upon phorbol ester-mediated PKC activation of OVCA433 cells, TJ strength is decreased and claudin-4 localization is altered. Analyses using PKC inhibitors and siRNA suggest that PKCε, an isoform typically expressed in ovarian cancer cells, may be important in the TPAmediated claudin-4 phosphorylation and weakening of the TJs. Furthermore, immunofluorescence studies showed that claudin-4 and PKCε are co-localized at the TJs in these cells. The modulation of claudin-4 activity by PKCε may not only provide a mechanism for disrupting TJ function in ovarian cancer, but may also be important in the regulation of TJ function in normal epithelial cells.

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“…Another reported target of PKA phosphorylation is claudin-16. In the study by Ikari et al (48), claudin-16 acted as a Mg 2ϩ -selective pore when heterologously expressed in MDCKII cells. This effect was associated with constitutive phosphorylation of claudin-16 and was reversed by adding inhibitors of PKA and adenylate cyclase, or by introducing the point mutation Ser217Ala.…”

Section: Phosphorylation Of Claudinsmentioning

confidence: 99%

“…As discussed in the previous section, the in vitro studies of claudin-16 permeability properties have yielded conflicting results, and the idea that claudin-16 by itself forms a Mg 2ϩ pore is not well supported by the available evidence (44,48,57). There are two alternative models that have been proposed to explain the pathogenesis of the disease.…”

Section: Role Of Claudins In Human Diseasesmentioning

confidence: 99%

Biology of claudins

Angelow¹,

Ahlstrom²,

Yu³

2008

American Journal of Physiology-Renal Physiology

304

294

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Claudins are a family of tight junction membrane proteins that regulate paracellular permeability of epithelia, likely by forming the lining of the paracellular pore. Claudins are expressed throughout the renal tubule, and mutations in two claudin genes are now known to cause familial hypercalciuric hypomagnesemia with nephrocalcinosis. In this review, we discuss recent advances in our understanding of the physiological role of various claudins in normal kidney function, and in understanding the fundamental biology of claudins, including the molecular basis for selectivity of permeation, claudin interactions in tight junction formation, and regulation of claudins by protein kinases and other intracellular signals.

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Phosphorylation of paracellin-1 at Ser217 by protein kinase A is essential for localization in tight junctions

Cited by 95 publications

References 57 publications

Claudin-4 forms paracellular chloride channel in the kidney and requires claudin-8 for tight junction localization

Claudin-4 forms paracellular chloride channel in the kidney and requires claudin-8 for tight junction localization

Phosphorylation of claudin-4 by PKCε regulates tight junction barrier function in ovarian cancer cells

Biology of claudins

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