title: Integrin-linked kinase modulates lipopolysaccharide- and Helicobacter pylori-induced nuclear factor κB-activated tumor necrosis factor-α production via regulation of p65 serine 536 phosphorylation.
abstract: Integrin-linked kinase (ILK) is a ubiquitously expressed and highly conserved serine-threonine protein kinase that regulates cellular responses to a wide variety of extracellular stimuli. ILK is involved in cell-matrix interactions, cytoskeletal organization, and cell signaling. ILK signaling has also been implicated in oncogenesis and progression of cancers. However, its role in the innate immune system remains unknown. Here, we show that ILK mediates pro-inflammatory signaling in response to lipopolysaccharide (LPS). Pharmacological or genetic inhibition of ILK in mouse embryonic fibroblasts and macrophages selectively blocks LPS-induced production of the pro-inflammatory cytokine tumor necrosis factor α (TNF-α). ILK is required for LPS-induced activation of nuclear factor κB (NF-κB) and transcriptional induction of TNF-α. The modulation of LPS-induced TNF-α synthesis by ILK does not involve the classical NF-κB pathway, because IκB-α degradation and p65 nuclear translocation are both unaffected by ILK inhibition. Instead, ILK is involved in an alternative activation of NF-κB signaling by modulating the phosphorylation of p65 at Ser-536. Furthermore, ILK-mediated alternative NF-κB activation through p65 Ser-536 phosphorylation also occurs during Helicobacter pylori infection in macrophages and gastric cancer cells. Moreover, ILK is required for H. pylori-induced TNF-α secretion in macrophages. Although ILK-mediated phosphorylation of p65 at Ser-536 is independent of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway during LPS stimulation, upon H. pylori infection this event is dependent on the PI3K/Akt pathway. Our findings implicate ILK as a critical regulatory molecule for the NF-κB-mediated pro-inflammatory signaling pathway, which is essential for innate immune responses against pathogenic microorganisms.
authors.affiliation: From the Centre for Cancer Research, MIMR-PHI Institute of Medical Research, and Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia.
authors.forename: Afsar U
authors.initials: AU
authors.lastname: Ahmed
authors.affiliation: From the Centre for Cancer Research, MIMR-PHI Institute of Medical Research, and Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia.
authors.forename: Soroush T
authors.initials: ST
authors.lastname: Sarvestani
authors.affiliation: From the Centre for Cancer Research, MIMR-PHI Institute of Medical Research, and Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Gantier
authors.affiliation: From the Centre for Cancer Research, MIMR-PHI Institute of Medical Research, and Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia bryan.williams@mimr-phi.org.
authors.forename: Bryan R G
authors.initials: BR
authors.lastname: Williams
authors.affiliation: From the Centre for Cancer Research, MIMR-PHI Institute of Medical Research, and Department of Molecular and Translational Science, Monash University, Clayton, Victoria 3168, Australia greg.hannigan@mimr-phi.org.
authors.forename: Gregory E
authors.initials: GE
authors.lastname: Hannigan
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D005786:Gene Expression Regulation; D016481:Helicobacter Infections / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D016480:Helicobacter pylori / Q000502:physiology*; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism*; D019892:eIF-2 Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000099200:Scaffold Protein ILK
chemical_list: D008070:Lipopolysaccharides; D016328:NF-kappa B; D017346:Protein Serine-Threonine Kinases; D012694:Serine; D014409:Tumor Necrosis Factor-alpha; D019892:eIF-2 Kinase; D000099200:Scaffold Protein ILK
keywords: Helicobacter pylori; Inflammation; Innate Immunity; Integrin-linked Kinase (ILK); Lipopolysaccharide (LPS); NF-kappa B; Phosphatidylinositide 3-Kinase (PI 3-Kinase); Small Molecule; Tumor Necrosis Factor (TNF); p65
references.citation: Annu Rev Pharmacol Toxicol. 1995;35:655-77
references.pmid: 7598511
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 15;101(24):8841-2
references.pmid: 15187230
references.citation: J Neurosci. 2008 Aug 13;28(33):8246-56
references.pmid: 18701687
references.citation: Acta Histochem. 2013 Mar;115(2):128-36
references.pmid: 22683084
references.citation: Carcinogenesis. 2007 Sep;28(9):1893-901
references.pmid: 17468516
references.citation: J Biol Chem. 2003 Aug 29;278(35):32552-60
references.pmid: 12807870
references.citation: Biochem Biophys Res Commun. 1999 Apr 29;258(1):17-20
references.pmid: 10222227
references.citation: J Immunol. 2009 Dec 15;183(12):8099-109
references.pmid: 20007577
references.citation: Nat Immunol. 2010 Aug;11(8):734-42
references.pmid: 20639876
references.citation: Mol Cell. 2009 Oct 9;36(1):131-40
references.pmid: 19818716
references.citation: Biochem J. 2006 May 15;396(1):193-200
references.pmid: 16472257
references.citation: J Biol Chem. 2011 Sep 30;286(39):34036-50
references.pmid: 21816815
references.citation: Nat Cell Biol. 2001 Oct;3(10):891-6
references.pmid: 11584270
references.citation: J Biol Chem. 1999 Oct 22;274(43):30353-6
references.pmid: 10521409
references.citation: World J Gastroenterol. 2013 Aug 21;19(31):5085-93
references.pmid: 23964142
references.citation: Cancer Biol Ther. 2011 May 1;11(9):826-38
references.pmid: 21358264
references.citation: Cell Microbiol. 2002 May;4(5):285-96
references.pmid: 12064285
references.citation: Annu Rev Immunol. 1989;7:625-55
references.pmid: 2540776
references.citation: J Biol Chem. 2006 May 12;281(19):13275-13284
references.pmid: 16513650
references.citation: J Cell Biol. 2008 Feb 25;180(4):681-9
references.pmid: 18283114
references.citation: Cancer Res. 2006 Jan 1;66(1):393-403
references.pmid: 16397254
references.citation: Immunity. 2010 Apr 23;32(4):518-30
references.pmid: 20362473
references.citation: Nat Rev Mol Cell Biol. 2006 Jan;7(1):20-31
references.pmid: 16493410
references.citation: J Immunol. 2010 Sep 15;185(6):3685-93
references.pmid: 20696864
references.citation: Oncogene. 2011 Feb 3;30(5):521-34
references.pmid: 20838383
references.citation: Eur J Immunol. 2013 Mar;43(3):779-92
references.pmid: 23310953
references.citation: Scand J Gastroenterol. 1997 Mar;32(3):203-11
references.pmid: 9085455
references.citation: Am J Physiol Cell Physiol. 2009 Oct;297(4):C814-22
references.pmid: 19587216
references.citation: J Biol Chem. 2005 Jun 24;280(25):23496-501
references.pmid: 15849198
references.citation: Oncogene. 2004 May 6;23(21):3883-7
references.pmid: 15021910
references.citation: Cancer Lett. 2012 Sep 28;322(2):133-8
references.pmid: 22459353
references.citation: J Biol Chem. 2002 Feb 1;277(5):3109-16
references.pmid: 11724787
references.citation: Biochem Biophys Res Commun. 2008 Mar 28;368(1):94-9
references.pmid: 18222170
references.citation: Circulation. 2006 Nov 21;114(21):2271-9
references.pmid: 17088456
references.citation: Immunol Rev. 2012 Mar;246(1):125-40
references.pmid: 22435551
references.citation: J Cell Sci. 2008 Oct 1;121(Pt 19):3121-32
references.pmid: 18799788
references.citation: Science. 2001 Aug 31;293(5535):1653-7
references.pmid: 11533489
references.citation: J Biol Chem. 2006 Sep 15;281(37):26976-84
references.pmid: 16840782
references.citation: Oncogene. 2006 Dec 14;25(59):7747-57
references.pmid: 16799642
references.citation: Nat Rev Cancer. 2005 Jan;5(1):51-63
references.pmid: 15630415
references.citation: J Immunol. 1998 May 1;160(9):4535-42
references.pmid: 9574560
references.citation: J Immunol. 2007 Nov 15;179(10):7121-7
references.pmid: 17982104
references.citation: Nat Immunol. 2008 Apr;9(4):361-8
references.pmid: 18297073
references.citation: Nature. 2007 Oct 18;449(7164):862-6
references.pmid: 17943123
references.citation: Immunol Rev. 2012 Mar;246(1):77-94
references.pmid: 22435548
references.citation: J Biol Chem. 2002 Sep 13;277(37):34223-8
references.pmid: 12091382
references.citation: J Exp Med. 2007 Nov 26;204(12):2963-76
references.pmid: 17998391
references.citation: J Biol Chem. 2001 Jun 1;276(22):18934-40
references.pmid: 11259436
references.citation: FASEB J. 2007 Sep;21(11):2642-54
references.pmid: 17431096
references.citation: Oncogene. 2011 Jun 16;30(24):2778-88
references.pmid: 21278793
references.citation: Nature. 1996 Jan 4;379(6560):91-6
references.pmid: 8538749
references.citation: Leuk Res. 2010 Oct;34(10):1358-65
references.pmid: 20193963
references.citation: Oncogene. 2010 Dec 2;29(48):6343-56
references.pmid: 20838384
references.citation: J Immunol. 2003 Jun 1;170(11):5630-5
references.pmid: 12759443
references.citation: Cell. 2006 Jun 2;125(5):943-55
references.pmid: 16751103
references.citation: J Immunol. 2007 Jun 15;178(12):7686-93
references.pmid: 17548605
references.citation: PLoS One. 2010 Aug 23;5(8):e12356
references.pmid: 20827300
references.citation: Proc Natl Acad Sci U S A. 2005 Jun 28;102(26):9300-5
references.pmid: 15972330
references.citation: J Biochem Mol Biol. 2006 Jan 31;39(1):1-8
references.pmid: 16466631
references.citation: Mol Cell Biol. 2001 Oct;21(20):7065-77
references.pmid: 11564889
references.citation: Adv Exp Med Biol. 2007;598:49-61
references.pmid: 17892204
references.citation: Annu Rev Immunol. 1995;13:437-57
references.pmid: 7542010
references.citation: Nat Immunol. 2004 Nov;5(11):1166-74
references.pmid: 15489856
references.citation: Inflamm Bowel Dis. 1999 May;5(2):119-33
references.pmid: 10338381
references.citation: Apoptosis. 2012 Dec;17(12):1261-74
references.pmid: 23054083
references.citation: Nat Immunol. 2004 May;5(5):495-502
references.pmid: 15107846
references.citation: Science. 1998 Dec 11;282(5396):2085-8
references.pmid: 9851930
references.citation: J Biol Chem. 2005 Oct 14;280(41):34538-47
references.pmid: 16105840
references.citation: Oncogene. 2011 Oct 27;30(43):4375-85
references.pmid: 21602880
references.citation: Breast Cancer Res. 2009;11(3):R25
references.pmid: 19409087
references.citation: J Immunol. 2001 Jan 1;166(1):574-81
references.pmid: 11123339
references.citation: Immunology. 2004 Oct;113(2):153-62
references.pmid: 15379975
references.citation: Matrix Biol. 2010 Apr;29(3):161-5
references.pmid: 20018240
references.citation: Immunol Rev. 2012 Mar;246(1):14-35
references.pmid: 22435545