Article thumbnail
Location of Repository

Ste20-Related Proline/Alanine-Rich Kinase (SPAK) Regulated Transcriptionally by Hyperosmolarity Is Involved in Intestinal Barrier Function

By Yutao Yan, Guillaume Dalmasso, Hang Thi Thu Nguyen, Tracy S. Obertone, Shanthi V. Sitaraman and Didier Merlin


The Ste20-related protein proline/alanine-rich kinase (SPAK) plays important roles in cellular functions such as cell differentiation and regulation of chloride transport, but its roles in pathogenesis of intestinal inflammation remain largely unknown. Here we report significantly increased SPAK expression levels in hyperosmotic environments, such as mucosal biopsy samples from patients with Crohn's disease, as well as colon tissues of C57BL/6 mice and Caco2-BBE cells treated with hyperosmotic medium. NF-κB and Sp1-binding sites in the SPAK TATA-less promoter are essential for SPAK mRNA transcription. Hyperosmolarity increases the ability of NF-κB and Sp1 to bind to their binding sites. Knock-down of either NF-κB or Sp1 by siRNA reduces the hyperosmolarity-induced SPAK expression levels. Furthermore, expression of NF-κB, but not Sp1, was upregulated by hyperosmolarity in vivo and in vitro. Nuclear run-on assays showed that hyperosmolarity increases SPAK expression levels at the transcriptional level, without affecting SPAK mRNA stability. Knockdown of SPAK expression by siRNA or overexpression of SPAK in cells and transgenic mice shows that SPAK is involved in intestinal permeability in vitro and in vivo. Together, our data suggest that SPAK, the transcription of which is regulated by hyperosmolarity, plays an important role in epithelial barrier function

Topics: Research Article
Publisher: Public Library of Science
OAI identifier:
Provided by: PubMed Central

Suggested articles


  1. (2003). A convenient rabbit model of ocular epithelium damage induced by osmotic dehydration.
  2. (1993). A cooperative interaction between NF-kappa B and Sp1 is required for HIV-1 enhancer activation.
  3. (1994). A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells.
  4. (2004). A porous defense: the leaky epithelial barrier in intestinal disease.
  5. (1986). Activation of the AIDS retrovirus promoter by the cellular transcription factor,
  6. (2003). Aldose reductase induced by hyperosmotic stress mediates cardiomyocyte apoptosis: differential effects of sorbitol and mannitol.
  7. (1993). Association between proto-oncoprotein Rel and TATA-binding protein mediates transcriptional activation by NF-kappa B.
  8. (2002). Association of the Ste20-like kinase (SLK) with the microtubule. Role in Rac1-mediated regulation of actin dynamics during cell adhesion and spreading.
  9. (1999). Both Erk and p38 kinases are necessary for cytokine gene transcription.
  10. (2002). Cation chloride cotransporters interact with the stress-related kinases Ste20-related proline-alanine-rich kinase (SPAK) and oxidative stress response 1 (OSR1).
  11. (1990). Cellular basis for defective electrolyte transport in inflamed human colon.
  12. (1996). Changes in cytoskeletal actin content, F-actin distribution, and surface morphology during HL-60 cell volume regulation.
  13. (2006). Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter.
  14. (2006). Circuitry of nuclear factor kappaB signaling.
  15. (2007). Cloning and characterization of a new intestinal inflammation-associated colonic epithelial Ste20-related protein kinase isoform.
  16. (1995). Cloning and characterization of seven cDNAs for hyperosmolarity-responsive (HOR) genes of Saccharomyces cerevisiae.
  17. (1982). Comparison of the composition of faecal fluid in Crohn’s disease and ulcerative colitis.
  18. (2002). Contribution of F-actin to barrier properties of the blood-joint pathway.
  19. (2001). Cytoskeletal regulation of pulmonary vascular permeability.
  20. (2000). Deficiency of a STE20/PAK family kinase LOK leads to the acceleration of LFA-1 clustering and cell adhesion of activated lymphocytes.
  21. (2000). Dehydration activates an NF-kappaB-driven, COX2-dependent survival mechanism in renal medullary interstitial cells.
  22. (1991). Developmental expression of Sp1 in the mouse.
  23. (1993). Differential regulation of vascular cell adhesion molecule 1 gene expression by specific NF-kappa B subunits in endothelial and epithelial cells.
  24. (1997). Dinarello CA
  25. (2007). Edaravone mimics sphingosine-1-phosphate-induced endothelial barrier enhancement in human microvascular endothelial cells.
  26. (1995). Endothelial interferon regulatory factor 1 cooperates with NF-kappa B as a transcriptional activator of vascular cell adhesion molecule 1.
  27. (1997). Expression of the bumetanide-sensitive Na-K-Cl cotransporter BSC2 is differentially regulated by fluid mechanical and inflammatory cytokine stimuli in vascular endothelium.
  28. (2007). Focal adhesion kinase controls pH-dependent epidermal barrier homeostasis by regulating actin-directed Na+/H+ exchanger 1 plasma membrane localization.
  29. (1998). Functional interference of Sp1 and NF-kappaB through the same DNA binding site.
  30. (1997). High glucose and hyperosmolarity increase secretion of interleukin-1 beta in cultured human aortic endothelial cells.
  31. (2005). Hyperosmolar saline is a proinflammatory stress on the mouse ocular surface.
  32. (2008). Hyperosmolarity causes inflammation through the methylation of protein phosphatase 2A. Inflamm Res.
  33. (2003). Hyperosmolarity enhances the lung capillary barrier.
  34. (2007). Hyperosmolarity-induced apoptosis in human corneal epithelial cells is mediated by cytochrome c and MAPK pathways.
  35. (1999). Hyperosmolarity-induced interleukin-8 expression in human bronchial epithelial cells through p38 mitogen-activated protein kinase.
  36. (2002). Hyperosmotic stimuli inhibit VCAM1 expression in cultured endothelial cells via effects on interferon regulatory factor-1 expression and activity.
  37. (2002). Hyperosmotic stress induces nuclear factor-kappaB activation and interleukin-8 production in human intestinal epithelial cells.
  38. (1995). Immunosuppression by glucocorticoids: inhibition of NF-kappa B activity through induction of I kappa B synthesis.
  39. (2007). Induction of vascular permeability: beta PIX and GIT1 scaffold the activation of extracellular signal-regulated kinase by PAK.
  40. (2003). Inhibition of interferon-gamma expression by osmotic shrinkage of peripheral blood lymphocytes.
  41. (1994). Inhibition of NF-kappa B by sodium salicylate and aspirin.
  42. (2004). Inhibition of p38 MAP kinase- and RICK/NF-kappaB-signaling suppresses inflammatory bowel disease.
  43. (2005). Integrative model of the response of yeast to osmotic shock.
  44. (1989). Intestinal permeability in patients with Crohn’s disease and their healthy relatives.
  45. (2006). JNK and ERK MAP kinases mediate induction of IL-1beta, TNF-alpha and IL-8 following hyperosmolar stress in human limbal epithelial cells.
  46. (2000). Kapus A
  47. (2001). Kusumi A
  48. (2004). Lavoinne A
  49. (2005). Luminal hyperosmolarity decreases Na transport and impairs barrier function of sheep rumen epithelium.
  50. (1993). Mechanism of initiator-mediated transcription: evidence for a functional interaction between the TATA-binding protein and DNA in the absence of a specific recognition sequence.
  51. (2007). Mice lacking NKCC1 are protected from development of bacteremia and hypothermic sepsis secondary to bacterial pneumonia.
  52. (1999). Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human.
  53. (2000). Mitogen-activated protein phosphorylation in endothelial cells exposed to hyperosmolar conditions.
  54. (2000). Modulation of CFTR gene expression in HT-29 cells by extracellular hyperosmolarity.
  55. (2007). Molecular pathogenesis of inflammatory bowel disease: genotypes, phenotypes and personalized medicine.
  56. (2005). Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters.
  57. (1988). Neonatal necrotizing enterocolitis. Inflammatory bowel disease of the newborn.
  58. (1998). NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses.
  59. (1993). NF-kappa B subunit-specific regulation of the interleukin-8 promoter.
  60. (1989). NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control.
  61. (2008). Nuclear factor-kappaB is a critical mediator of Ste20-like proline-/ alanine-rich kinase regulation in intestinal inflammation.
  62. (1988). Organic anions and the diarrhea of inflammatory bowel disease.
  63. (1988). Osmoregulation by slow changes in aldose reductase and rapid changes in sorbitol flux.
  64. (2004). Osmoregulation of taurine transporter function and expression in retinal pigment epithelial, ganglion, and muller cells.
  65. (1989). Osmotic regulation of aldose reductase protein synthesis in renal medullary cells.
  66. (2004). p21-activated kinase regulates endothelial permeability through modulation of contractility.
  67. (1998). p38 MAPK is required for CD40-induced gene expression and proliferation in B lymphocytes.
  68. (1999). Pak to the future.
  69. (2003). PASK (proline-alanine-rich STE20-related kinase), a regulatory kinase of the Na-K-Cl cotransporter (NKCC1).
  70. (1997). Post-transcriptional regulation contributes to Drosophila clock gene mRNA cycling.
  71. (2003). Proinflammatory cytokines disrupt epithelial barrier function by apoptosisindependent mechanisms.
  72. (2006). Proteasome-mediated degradation of IkappaBalpha and processing of p105 in Crohn disease and ulcerative colitis.
  73. (1986). Purification and biochemical characterization of the promoter-specific transcription factor,
  74. (2000). Reactive oxygen intermediates are involved in IL-8 production induced by hyperosmotic stress in human bronchial epithelial cells.
  75. (1980). Ribonucleic acid synthesis in embryonic chick muscle, rates of synthesis and half-lives of transfer and ribosomal RNA species.
  76. (1982). Role of endothelial cell cytoskeleton in control of endothelial permeability.
  77. (1998). Role of MEKK1 in cell survival and activation of JNK and ERK pathways defined by targeted gene disruption.
  78. (1997). Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis.
  79. (1995). Role of transcriptional activation of I kappa B alpha in mediation of immunosuppression by glucocorticoids.
  80. (2006). Selective ablation of matrix metalloproteinase-2 exacerbates experimental colitis: contrasting role of gelatinases in the pathogenesis of colitis.
  81. (1985). Short-term stimulation of lipogenesis by triiodothyronine in maintenance cultures of rat hepatocytes.
  82. (1998). Signal transduction by the c-Jun N-terminal kinase (JNK)–from inflammation to development.
  83. (1999). Signaling by the germinal center kinase family of protein kinases.
  84. (2004). Signaling to NF-kappaB.
  85. (1994). Sp1 is a critical factor for the monocytic specific expression of human CD14.
  86. (2004). SPAK kinase is a substrate and target of PKCtheta in T-cell receptor-induced AP-1 activation pathway.
  87. (2000). SPAK, a STE20/SPS1-related kinase that activates the p38 pathway.
  88. (2004). Stimulation of matrix metalloproteinases by hyperosmolarity via a JNK pathway in human corneal epithelial cells.
  89. (2004). Sturm A
  90. (2001). The hepatitis B virus X protein induces HIV-1 replication and transcription in synergy with T-cell activation signals: functional roles of NFkappaB/NF-AT and SP1-binding sites in the HIV-1 long terminal repeat promoter.
  91. (1989). The involvement of NFkappa B in beta-interferon gene regulation reveals its role as widely inducible mediator of signal transduction.
  92. (2001). Toll-like receptor-mediated NF-kappaB activation: a phylogenetically conserved paradigm in innate immunity.
  93. (1999). Transcriptional activation by NF-kappaB requires multiple coactivators.
  94. (2005). Transforming growth factor-beta regulation of epithelial tight junction proteins enhances barrier function and blocks enterohemorrhagic Escherichia coli O157:H7-induced increased permeability.
  95. (1995). Triggering of the human interleukin-6 gene by interferon-gamma and tumor necrosis factor-alpha in monocytic cells involves cooperation between interferon regulatory factor-1, NF kappa B, and Sp1 transcription factors.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.