Spatially restricted activation of RhoA signalling at epithelial junctions by p114RhoGEF drives junction formation and morphogenesis.
Signalling by the GTPase RhoA, a key regulator of epithelial cell behaviour, can stimulate opposing processes: RhoA can promote junction formation and apical constriction, and reduce adhesion and cell spreading. Molecular mechanisms are thus required that ensure spatially restricted and process-specific RhoA activation. For many fundamental processes, including assembly of the epithelial junctional complex, such mechanisms are still unknown. Here we show that p114RhoGEF is a junction-associated protein that drives RhoA signalling at the junctional complex and regulates tight-junction assembly and epithelial morphogenesis. p114RhoGEF is required for RhoA activation at cell-cell junctions, and its depletion stimulates non-junctional Rho signalling and induction of myosin phosphorylation along the basal domain. Depletion of GEF-H1, a RhoA activator inhibited by junctional recruitment, does not reduce junction-associated RhoA activation. p114RhoGEF associates with a complex containing myosin II, Rock II and the junctional adaptor cingulin, indicating that p114RhoGEF is a component of a junction-associated Rho signalling module that drives spatially restricted activation of RhoA to regulate junction formation and epithelial morphogenesis.
Department of Cell Biology, UCL Institute of Ophthalmology, University College London, Bath Street, London, UK.
Stephen J
SJ
Terry
Ceniz
C
Zihni
Ahmed
A
Elbediwy
Elisa
E
Vitiello
Isabelle V
IV
Leefa Chong San
Maria S
MS
Balda
Karl
K
Matter
D002460:Cell Line; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D064067:Rho Guanine Nucleotide Exchange Factors; D015398:Signal Transduction / Q000502:physiology*; D019108:Tight Junctions / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
C577230:ARHGEF18 protein, human; D020662:Guanine Nucleotide Exchange Factors; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D064067:Rho Guanine Nucleotide Exchange Factors; D020742:rhoA GTP-Binding Protein
Science. 2005 Mar 11;307(5715):1603-9
15761148
J Cell Biol. 2008 Nov 17;183(4):625-33
19001128
Nature. 2002 Dec 12;420(6916):629-35
12478284
J Cell Biol. 2000 May 15;149(4):775-82
10811819
Nat Cell Biol. 2002 Apr;4(4):294-301
11912491
BMC Cell Biol. 2009 Dec 22;10:95
20028514
Methods. 2003 Jul;30(3):228-34
12798137
J Biol Chem. 2003 Apr 18;278(16):13615-8
12606561
J Biol Chem. 2009 Apr 24;284(17):11454-66
19261619
Oncogene. 2005 Jan 6;24(1):65-76
15558029
J Cell Sci. 2005 Oct 15;118(Pt 20):4667-78
16188940
J Cell Sci. 2010 Jun 1;123(Pt 11):1841-50
20484664
Circ Res. 2003 Oct 31;93(9):848-56
14512443
Nat Rev Mol Cell Biol. 2008 Sep;9(9):690-701
18719708
Mol Biol Cell. 2006 Aug;17(8):3569-77
16723500
Trends Cell Biol. 2010 Mar;20(3):142-9
20061152
Mol Cell Biol. 1999 Feb;19(2):1334-45
9891067
Trends Cell Biol. 2008 May;18(5):210-9
18394899
J Biol Chem. 2001 Sep 7;276(36):33305-8
11457821
J Cell Biol. 2007 Jul 30;178(3):517-27
17646397
J Cell Biol. 1997 Jun 16;137(6):1421-31
9182672
J Cell Biol. 2003 Jul 21;162(2):223-32
12860967
EMBO Rep. 2009 Oct;10(10):1125-31
19730435
Curr Opin Cell Biol. 2002 Oct;14(5):546-56
12231348
Nat Rev Mol Cell Biol. 2005 Feb;6(2):167-80
15688002
Physiology (Bethesda). 2010 Feb;25(1):16-26
20134025
J Biol Chem. 1989 Aug 5;264(22):13131-9
2526812
J Cell Biol. 2003 Mar 3;160(5):729-40
12604587
Biochem J. 2000 Dec 1;352 Pt 2:319-25
11085924
J Cell Biol. 1999 Dec 27;147(7):1569-82
10613913
Mol Cell Biol. 2006 Mar;26(6):2387-98
16508013
Trends Cell Biol. 2003 Apr;13(4):169-76
12667754
Mol Biol Cell. 2007 Sep;18(9):3429-39
17596509
J Cell Biol. 1996 Aug;134(4):1031-49
8769425
Nat Rev Mol Cell Biol. 2004 May;5(5):355-66
15122349
Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10629-33
7479854
Mol Biol Cell. 2009 Dec;20(24):5074-85
19828736
Dev Cell. 2005 May;8(5):777-86
15866167