title: Novel role for p21-activated kinase 2 in thrombin-induced monocyte migration.
abstract: To understand the role of thrombin in inflammation, we tested its effects on migration of THP-1 cells, a human monocytic cell line. Thrombin induced THP-1 cell migration in a dose-dependent manner. Thrombin induced tyrosine phosphorylation of Pyk2, Gab1, and p115 RhoGEF, leading to Rac1- and RhoA-dependent Pak2 activation. Downstream to Pyk2, Gab1 formed a complex with p115 RhoGEF involving their pleckstrin homology domains. Furthermore, inhibition or depletion of Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, or Pak2 levels substantially attenuated thrombin-induced THP-1 cell F-actin cytoskeletal remodeling and migration. Inhibition or depletion of PAR1 also blocked thrombin-induced activation of Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, and Pak2, resulting in diminished THP-1 cell F-actin cytoskeletal remodeling and migration. Similarly, depletion of Gα12 negated thrombin-induced Pyk2, Gab1, p115 RhoGEF, Rac1, RhoA, and Pak2 activation, leading to attenuation of THP-1 cell F-actin cytoskeletal remodeling and migration. These novel observations reveal that thrombin induces monocyte/macrophage migration via PAR1-Gα12-dependent Pyk2-mediated Gab1 and p115 RhoGEF interactions, leading to Rac1- and RhoA-targeted Pak2 activation. Thus, these findings provide mechanistic evidence for the role of thrombin and its receptor PAR1 in inflammation.
authors.affiliation: From the Department of Physiology, University of Tennessee Health Science Center, Memphis, Tennessee 38163.
authors.forename: Ravisekhar
authors.initials: R
authors.lastname: Gadepalli
authors.forename: Sivareddy
authors.initials: S
authors.lastname: Kotla
authors.forename: Mark R
authors.initials: MR
authors.lastname: Heckle
authors.forename: Shailendra K
authors.initials: SK
authors.lastname: Verma
authors.forename: Nikhlesh K
authors.initials: NK
authors.lastname: Singh
authors.forename: Gadiparthi N
authors.initials: GN
authors.lastname: Rao
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology; D051418:Focal Adhesion Kinase 2 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009000:Monocytes / Q000166:cytology / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D064067:Rho Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism; D013917:Thrombin / Q000235:genetics / Q000378:metabolism*; D054462:p21-Activated Kinases / Q000235:genetics / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; C098587:GAB1 protein, human; C490897:Gab1 protein, mouse; D009479:Neuropeptides; D010750:Phosphoproteins; C107953:RAC1 protein, human; C484602:Rac1 protein, mouse; D064067:Rho Guanine Nucleotide Exchange Factors; C066737:RHOA protein, human; D051418:Focal Adhesion Kinase 2; C103547:PTK2B protein, human; C496053:Ptk2b protein, mouse; C515907:PAK2 protein, human; C515909:Pak2 protein, mouse; D054462:p21-Activated Kinases; D013917:Thrombin; C526380:RhoA protein, mouse; D020830:rac1 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein
keywords: G Protein-coupled Receptors (GPCRs); Inflammation; Macrophages; Migration; Signaling
references.citation: J Biol Chem. 2012 Jul 20;287(30):25490-500
references.pmid: 22661716
references.citation: J Thromb Haemost. 2005 Aug;3(8):1800-14
references.pmid: 16102047
references.citation: Annu Rev Biochem. 2003;72:743-81
references.pmid: 12676796
references.citation: J Biol Chem. 2003 Jul 4;278(27):24644-50
references.pmid: 12714602
references.citation: Arterioscler Thromb Vasc Biol. 2011 May;31(5):1084-92
references.pmid: 21372295
references.citation: Oncogene. 2001 Mar 26;20(13):1570-81
references.pmid: 11313904
references.citation: Science. 1997 Jan 24;275(5299):533-6
references.pmid: 8999798
references.citation: J Clin Invest. 2009 Jan;119(1):136-45
references.pmid: 19065049
references.citation: Mol Cell Biol. 1999 Mar;19(3):1784-99
references.pmid: 10022866
references.citation: Mol Biol Cell. 2012 Nov;23(21):4212-25
references.pmid: 22956765
references.citation: Thromb Haemost. 2008 Feb;99(2):305-15
references.pmid: 18278179
references.citation: Curr Pharm Des. 2003;9(28):2349-65
references.pmid: 14529396
references.citation: Pharmacol Rev. 2001 Jun;53(2):245-82
references.pmid: 11356985
references.citation: Proc Natl Acad Sci U S A. 2003 Jan 21;100(2):733-8
references.pmid: 12515866
references.citation: J Pharmacol Exp Ther. 2003 Nov;307(2):437-42
references.pmid: 12966163
references.citation: J Biol Chem. 2004 Oct 8;279(41):43273-84
references.pmid: 15292195
references.citation: Circ Res. 2009 Apr 24;104(8):943-51
references.pmid: 19265041
references.citation: J Biol Chem. 2007 Dec 7;282(49):35666-78
references.pmid: 17932039
references.citation: Mol Biol Cell. 2012 Jun;23(11):2076-91
references.pmid: 22496418
references.citation: Trends Cell Biol. 2003 Mar;13(3):122-30
references.pmid: 12628344
references.citation: J Biol Chem. 1996 Oct 11;271(41):25452-8
references.pmid: 8810315
references.citation: J Biol Chem. 2013 Apr 26;288(17):12232-43
references.pmid: 23467409
references.citation: Nature. 2002 Dec 12;420(6916):629-35
references.pmid: 12478284
references.citation: J Clin Invest. 1992 Oct;90(4):1614-21
references.pmid: 1328304
references.citation: J Biol Chem. 2000 Nov 17;275(46):35807-13
references.pmid: 10964909
references.citation: J Biol Chem. 2009 Sep 4;284(36):24133-43
references.pmid: 19574218
references.citation: Mol Cell Biol. 2009 Jun;29(11):3018-32
references.pmid: 19289496
references.citation: Arterioscler Thromb Vasc Biol. 2008 Oct;28(10):1844-50
references.pmid: 18617644
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 19;104(25):10619-24
references.pmid: 17537919
references.citation: Mol Cancer Res. 2004 Jul;2(7):395-402
references.pmid: 15280447
references.citation: Blood. 1998 Aug 15;92(4):1259-67
references.pmid: 9694714
references.citation: Circ Res. 1999 May 28;84(10):1186-93
references.pmid: 10347093
references.citation: Blood. 2001 Aug 1;98(3):667-73
references.pmid: 11468165
references.citation: J Biol Chem. 1998 Aug 14;273(33):21194-202
references.pmid: 9694876
references.citation: J Biol Chem. 2010 May 7;285(19):14259-66
references.pmid: 20197276
references.citation: J Cell Biol. 2006 Jul 31;174(3):437-45
references.pmid: 16864657
references.citation: Circ Res. 2003 Aug 8;93(3):221-9
references.pmid: 12855672
references.citation: J Neurosci. 2008 Jan 16;28(3):732-6
references.pmid: 18199772
references.citation: Trends Cell Biol. 2007 Jan;17(1):36-43
references.pmid: 17126549
references.citation: Blood. 1991 Jul 15;78(2):423-34
references.pmid: 2070078
references.citation: J Biol Chem. 2002 Nov 29;277(48):46791-8
references.pmid: 12244099
references.citation: Biochem J. 1992 Sep 15;286 ( Pt 3):945-50
references.pmid: 1417754
references.citation: J Cell Biol. 2000 Sep 18;150(6):1375-84
references.pmid: 10995442
references.citation: Nat Rev Mol Cell Biol. 2005 Feb;6(2):167-80
references.pmid: 15688002
references.citation: Am J Physiol Cell Physiol. 2006 Jan;290(1):C172-82
references.pmid: 16148030
references.citation: Cell. 2004 Jan 23;116(2):167-79
references.pmid: 14744429
references.citation: J Cell Sci. 2004 Sep 1;117(Pt 19):4343-54
references.pmid: 15331659
references.citation: Nat Rev Cardiol. 2010 Feb;7(2):77-86
references.pmid: 20065951
references.citation: Nature. 2011 May 19;473(7347):317-25
references.pmid: 21593864
references.citation: Cancer Res. 2011 Nov 1;71(21):6561-6
references.pmid: 22009534
references.citation: N Engl J Med. 2005 Apr 21;352(16):1685-95
references.pmid: 15843671
references.citation: EMBO J. 2012 Jul 18;31(14):3118-29
references.pmid: 22728827
references.citation: Mol Biol Cell. 2002 Jun;13(6):2132-46
references.pmid: 12058075
references.citation: J Biol Chem. 2000 Aug 11;275(32):24470-6
references.pmid: 10821841
references.citation: Mol Biol Cell. 2003 Apr;14(4):1691-708
references.pmid: 12686619
references.citation: Mol Pharmacol. 2003 Nov;64(5):1199-209
references.pmid: 14573770
references.citation: J Biol Chem. 1996 Aug 30;271(35):20997-1000
references.pmid: 8798379
references.citation: J Biol Chem. 2006 Aug 11;281(32):23285-95
references.pmid: 16787925
references.citation: Circulation. 2011 Feb 1;123(4):e18-e209
references.pmid: 21160056
references.citation: J Biol Chem. 2006 Jul 21;281(29):19925-38
references.pmid: 16687399
references.citation: Mol Biol Cell. 2009 Nov;20(21):4596-610
references.pmid: 19776354
references.citation: Blood. 2001 Jan 15;97(2):359-66
references.pmid: 11154209
references.citation: J Cell Sci Suppl. 1988;9:169-84
references.pmid: 2855803
references.citation: Circ Res. 2009 May 8;104(9):1066-75
references.pmid: 19359598
references.citation: Arterioscler Thromb Vasc Biol. 2012 Nov;32(11):2652-61
references.pmid: 22922962
references.citation: Mol Cell Biol. 1999 Mar;19(3):1881-91
references.pmid: 10022875