title: Rap1 potentiates endothelial cell junctions by spatially controlling myosin II activity and actin organization.
abstract: Reorganization of the actin cytoskeleton is responsible for dynamic regulation of endothelial cell (EC) barrier function. Circumferential actin bundles (CAB) promote formation of linear adherens junctions (AJs) and tightening of EC junctions, whereas formation of radial stress fibers (RSF) connected to punctate AJs occurs during junction remodeling. The small GTPase Rap1 induces CAB formation to potentiate EC junctions; however, the mechanism underlying Rap1-induced CAB formation remains unknown. Here, we show that myotonic dystrophy kinase-related CDC42-binding kinase (MRCK)-mediated activation of non-muscle myosin II (NM-II) at cell-cell contacts is essential for Rap1-induced CAB formation. Our data suggest that Rap1 induces FGD5-dependent Cdc42 activation at cell-cell junctions to locally activate the NM-II through MRCK, thereby inducing CAB formation. We further reveal that Rap1 suppresses the NM-II activity stimulated by the Rho-ROCK pathway, leading to dissolution of RSF. These findings imply that Rap1 potentiates EC junctions by spatially controlling NM-II activity through activation of the Cdc42-MRCK pathway and suppression of the Rho-ROCK pathway.
authors.affiliation: Department of Cell Biology, National Cerebral and Cardiovascular Center Research Institute, Suita, Osaka 565-8565, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Ando
authors.forename: Shigetomo
authors.initials: S
authors.lastname: Fukuhara
authors.forename: Takahiro
authors.initials: T
authors.lastname: Moriya
authors.forename: Yutaro
authors.initials: Y
authors.lastname: Obara
authors.forename: Norimichi
authors.initials: N
authors.lastname: Nakahata
authors.forename: Naoki
authors.initials: N
authors.lastname: Mochizuki
mesh_terms: D000199:Actins / Q000378:metabolism*; D015153:Blotting, Western; D002448:Cell Adhesion; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D007365:Intercellular Junctions / Q000378:metabolism*; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D065636:Myotonin-Protein Kinase; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020764:cdc42 GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D034741:RNA, Small Interfering; D000089804:Shelterin Complex; C440809:TERF2IP protein, human; D034501:Telomere-Binding Proteins; C118666:CDC42BPB protein, human; D011505:Protein-Tyrosine Kinases; D065636:Myotonin-Protein Kinase; D054460:rho-Associated Kinases; D024462:Myosin Type II; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins
references.citation: Mol Cell Biol. 2005 Jan;25(1):136-46
references.pmid: 15601837
references.citation: J Exp Med. 2010 Apr 12;207(4):881-96
references.pmid: 20308363
references.citation: Mol Biol Cell. 2005 May;16(5):2207-17
references.pmid: 15728722
references.citation: Nat Rev Mol Cell Biol. 2012 Oct;13(10):673-9
references.pmid: 22931853
references.citation: J Biol Chem. 1985 Aug 25;260(18):10027-31
references.pmid: 3839510
references.citation: Mol Biol Cell. 2006 Feb;17(2):966-76
references.pmid: 16339077
references.citation: J Clin Invest. 2001 Sep;108(5):689-701
references.pmid: 11544274
references.citation: Cell. 2008 Oct 3;135(1):123-36
references.pmid: 18854160
references.citation: Nat Cell Biol. 2005 Mar;7(3):255-61
references.pmid: 15723050
references.citation: Trends Cardiovasc Med. 2007 Jul;17(5):151-6
references.pmid: 17574122
references.citation: Nat Rev Mol Cell Biol. 2009 Mar;10(3):165-77
references.pmid: 19234476
references.citation: J Biol Chem. 2004 Nov 5;279(45):46621-30
references.pmid: 15333633
references.citation: J Biol Chem. 2010 Apr 16;285(16):11760-4
references.pmid: 20181950
references.citation: Mol Cell Biol. 1998 Jan;18(1):130-40
references.pmid: 9418861
references.citation: Cell Signal. 2011 Dec;23(12):2056-64
references.pmid: 21840392
references.citation: J Cell Sci. 2008 Jul 1;121(Pt 13):2115-22
references.pmid: 18565824
references.citation: Science. 2002 Feb 15;295(5558):1285-8
references.pmid: 11847339
references.citation: J Biol Chem. 2004 Aug 20;279(34):35127-32
references.pmid: 15166221
references.citation: FEBS Lett. 2000 Nov 24;485(2-3):183-8
references.pmid: 11094164
references.citation: Exp Cell Res. 1999 Aug 1;250(2):576-80
references.pmid: 10413610
references.citation: Nat Cell Biol. 2006 Nov;8(11):1223-34
references.pmid: 17060906
references.citation: J Cell Biol. 2006 Oct 9;175(1):135-46
references.pmid: 17015620
references.citation: Mol Cell Biol. 2001 Apr;21(8):2767-78
references.pmid: 11283256
references.citation: J Cell Biol. 2011 Aug 22;194(4):643-56
references.pmid: 21844208
references.citation: Arterioscler Thromb Vasc Biol. 2012 Nov;32(11):2694-701
references.pmid: 22922960
references.citation: Mol Biol Cell. 2011 Dec;22(23):4647-56
references.pmid: 21976697
references.citation: Nat Cell Biol. 2007 Dec;9(12):1392-400
references.pmid: 18037882
references.citation: J Cell Biol. 2004 Aug 2;166(3):393-405
references.pmid: 15277544
references.citation: FEBS Lett. 2011 May 6;585(9):1260-8
references.pmid: 21457715
references.citation: J Biol Chem. 2005 Jan 7;280(1):815-25
references.pmid: 15504743
references.citation: Circulation. 2012 Jun 26;125(25):3142-58
references.pmid: 22661514
references.citation: J Cell Biol. 2012 Mar 5;196(5):641-52
references.pmid: 22391038
references.citation: Mol Cell Biol. 2004 Aug;24(15):6690-700
references.pmid: 15254236
references.citation: J Cell Biol. 2007 Oct 22;179(2):247-54
references.pmid: 17954608
references.citation: BMC Biol. 2010 Feb 02;8:11
references.pmid: 20122254
references.citation: J Muscle Res Cell Motil. 1998 Nov;19(8):839-54
references.pmid: 10047984
references.citation: Nat Methods. 2009 May;6(5):363-9
references.pmid: 19337237
references.citation: Nat Cell Biol. 2010 Jul;12(7):696-702
references.pmid: 20543839
references.citation: Blood. 2005 Mar 1;105(5):1950-5
references.pmid: 15374886
references.citation: J Physiol. 2003 Jun 1;549(Pt 2):489-500
references.pmid: 12692179
references.citation: Science. 1999 Mar 26;283(5410):2083-5
references.pmid: 10092231
references.citation: Nat Rev Mol Cell Biol. 2009 Nov;10(11):778-90
references.pmid: 19851336
references.citation: Cell. 2005 May 6;121(3):451-63
references.pmid: 15882626
references.citation: FEBS Lett. 2005 Sep 12;579(22):4966-72
references.pmid: 16115630
references.citation: Mol Biol Cell. 2003 May;14(5):1745-56
references.pmid: 12802051
references.citation: Curr Biol. 2009 Apr 28;19(8):668-74
references.pmid: 19345098
references.citation: Dev Cell. 2012 Jan 17;22(1):158-71
references.pmid: 22209329
references.citation: Am J Physiol Lung Cell Mol Physiol. 2008 Aug;295(2):L363-9
references.pmid: 18515405
references.citation: J Cell Biol. 2005 Aug 1;170(3):443-53
references.pmid: 16043513
references.citation: J Cell Sci. 2000 Feb;113 ( Pt 3):471-82
references.pmid: 10639334
references.citation: Trends Cell Biol. 2011 Sep;21(9):499-505
references.pmid: 21763139
references.citation: Nat Cell Biol. 2008 May;10(5):513-26
references.pmid: 18425120
references.citation: Arterioscler Thromb Vasc Biol. 2012 Apr;32(4):988-96
references.pmid: 22328776
references.citation: Mol Biol Cell. 2010 Feb 15;21(4):584-96
references.pmid: 20032304
references.citation: J Biochem Mol Biol. 2006 Mar 31;39(2):132-9
references.pmid: 16584626
references.citation: Mol Biol Cell. 2012 Jan;23(2):310-23
references.pmid: 22090347
references.citation: PLoS One. 2009 Nov 30;4(11):e8027
references.pmid: 19956566
references.citation: J Biol Chem. 2005 Mar 25;280(12):11675-82
references.pmid: 15661741
references.citation: Am J Pathol. 2008 Feb;172(2):521-33
references.pmid: 18187566