title: Involvement of p114-RhoGEF and Lfc in Wnt-3a- and dishevelled-induced RhoA activation and neurite retraction in N1E-115 mouse neuroblastoma cells.
abstract: The Wnt-induced planar cell polarity (PCP) signaling pathway is essential for polarized cell migration and morphogenesis. Dishevelled (Dvl) and its binding protein Daam1 mediate RhoA activation in this pathway. WGEF, a member of the Rho-guanine nucleotide exchange factor (Rho-GEF) family, was shown to play a role in Wnt-induced RhoA activation in Xenopus embryos. However, it has remained unknown which member(s) of a Rho-GEF family are involved in Wnt/Dvl-induced RhoA activation in mammalian cells. Here we identified p114-RhoGEF and Lfc (also called GEF-H1) as the Rho-GEFs responsible for Wnt-3a-induced RhoA activation in N1E-115 mouse neuroblastoma cells. We screened for Rho-GEF-silencing short-hairpin RNAs (shRNAs) that are capable of suppressing Dvl-induced neurite retraction in N1E-115 cells and found that p114-RhoGEF and Lfc shRNAs, but not WGEF shRNA, suppressed Dvl- and Wnt-3a-induced neurite retraction. p114-RhoGEF and Lfc shRNAs also inhibited Dvl- and Wnt-3a-induced RhoA activation, and p114-RhoGEF and Lfc proteins were capable of binding to Dvl and Daam1. Additionally, the Dvl-binding domains of p114-RhoGEF and Lfc inhibited Dvl-induced neurite retraction. Our results suggest that p114-RhoGEF and Lfc are critically involved in Wnt-3a- and Dvl-induced RhoA activation and neurite retraction in N1E-115 cells.
authors.affiliation: Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Takuji
authors.initials: T
authors.lastname: Tsuji
authors.forename: Yusaku
authors.initials: Y
authors.lastname: Ohta
authors.forename: Yuya
authors.initials: Y
authors.lastname: Kanno
authors.forename: Kenzo
authors.initials: K
authors.lastname: Hirose
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Ohashi
authors.forename: Kensaku
authors.initials: K
authors.lastname: Mizuno
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D045744:Cell Line, Tumor; D000072261:Dishevelled Proteins; D004789:Enzyme Activation / Q000187:drug effects; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D006801:Humans; D008246:Lysophospholipids / Q000494:pharmacology; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D016501:Neurites / Q000201:enzymology*; D009447:Neuroblastoma / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D025941:Protein Interaction Mapping; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D064067:Rho Guanine Nucleotide Exchange Factors; D051153:Wnt Proteins / Q000378:metabolism*; D060508:Wnt3 Protein; D060509:Wnt3A Protein; D029867:Xenopus Proteins; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C577402:Arhgef2 protein, mouse; C000606586:DVL1 protein, Xenopus; D000072261:Dishevelled Proteins; C000597170:GEF-H1 protein, Xenopus; D020662:Guanine Nucleotide Exchange Factors; D008246:Lysophospholipids; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D064067:Rho Guanine Nucleotide Exchange Factors; C555506:WNT3A protein, Xenopus; C555510:WNT3A protein, human; D051153:Wnt Proteins; D060508:Wnt3 Protein; D060509:Wnt3A Protein; C555511:Wnt3a protein, mouse; D029867:Xenopus Proteins; C500365:Daam1 protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; C032881:lysophosphatidic acid
references.citation: Nat Rev Genet. 2007 Feb;8(2):126-38
references.pmid: 17230199
references.citation: Circ Res. 2003 Oct 31;93(9):848-56
references.pmid: 14512443
references.citation: Trends Cell Biol. 2009 Mar;19(3):119-29
references.pmid: 19208479
references.citation: Nat Cell Biol. 2002 Apr;4(4):294-301
references.pmid: 11912491
references.citation: Cell. 2001 Dec 28;107(7):843-54
references.pmid: 11779461
references.citation: Genes Dev. 2002 Jul 1;16(13):1587-609
references.pmid: 12101119
references.citation: J Cell Biol. 1994 Aug;126(3):801-10
references.pmid: 8045941
references.citation: Mol Cell Biol. 2002 Feb;22(3):774-83
references.pmid: 11784854
references.citation: Nat Rev Mol Cell Biol. 2005 Feb;6(2):167-80
references.pmid: 15688002
references.citation: Bioessays. 2007 Feb;29(2):120-32
references.pmid: 17226800
references.citation: J Biol Chem. 2000 Feb 4;275(5):3577-82
references.pmid: 10652353
references.citation: J Biol Chem. 2002 Nov 8;277(45):43115-20
references.pmid: 12183458
references.citation: Dev Cell. 2002 May;2(5):525-35
references.pmid: 12015961
references.citation: Nat Methods. 2005 Dec;2(12):967-73
references.pmid: 16299483
references.citation: FEBS Lett. 2000 Nov 24;485(2-3):183-8
references.pmid: 11094164
references.citation: Development. 2005 Oct;132(20):4421-36
references.pmid: 16192308
references.citation: Mol Cell Biol. 2004 May;24(10):4487-501
references.pmid: 15121866
references.citation: J Biol Chem. 1998 Dec 25;273(52):34954-60
references.pmid: 9857026
references.citation: Proc Natl Acad Sci U S A. 2008 Jan 8;105(1):210-5
references.pmid: 18162551
references.citation: Nature. 2002 Dec 12;420(6916):629-35
references.pmid: 12478284
references.citation: EMBO J. 2008 Feb 20;27(4):606-17
references.pmid: 18256687
references.citation: J Biol Chem. 1999 Jul 23;274(30):21464-70
references.pmid: 10409711
references.citation: Dev Cell. 2007 May;12(5):699-712
references.pmid: 17488622
references.citation: Nature. 1997 May 15;387(6630):292-5
references.pmid: 9153394
references.citation: Mol Cell Biol. 1997 Mar;17(3):1201-11
references.pmid: 9032247
references.citation: Dev Biol. 2003 Jan 1;253(1):1-17
references.pmid: 12490194
references.citation: Genes Dev. 2007 Jun 15;21(12):1478-83
references.pmid: 17575049
references.citation: J Biol Chem. 2005 May 13;280(19):19358-63
references.pmid: 15755723
references.citation: Trends Genet. 1998 Nov;14(11):452-8
references.pmid: 9825673
references.citation: Annu Rev Cell Dev Biol. 2004;20:781-810
references.pmid: 15473860
references.citation: Neuron. 2005 Jul 7;47(1):85-100
references.pmid: 15996550
references.citation: Biochem J. 2000 Dec 1;352 Pt 2:319-25
references.pmid: 11085924
references.citation: Nat Genet. 2004 Feb;36(2):190-6
references.pmid: 14704669
references.citation: Genes Dev. 2003 Jan 15;17(2):295-309
references.pmid: 12533515
references.citation: EMBO J. 1999 Feb 1;18(3):578-85
references.pmid: 9927417
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 17;99(19):12085-90
references.pmid: 12196628
references.citation: Dev Cell. 2003 Sep;5(3):367-77
references.pmid: 12967557
references.citation: Mol Cell Biol. 2009 Nov;29(21):5963-73
references.pmid: 19667072
references.citation: Cell. 2001 Apr 6;105(1):81-91
references.pmid: 11301004
references.citation: Genes Cells. 1998 Oct;3(10):659-70
references.pmid: 9893023