Many cells in a developing embryo, including neurons and their axons and growth cones, must integrate multiple guidance cues to undergo directed growth and migration. The UNC-6/netrin, SLT-1/slit, and VAB-2/Ephrin guidance cues, and their receptors, UNC-40/DCC, SAX-3/Robo, and VAB-1/Eph, are known to be major regulators of cellular growth and migration. One important area of research is identifying the molecules that interpret this guidance information downstream of the guidance receptors to reorganize the actin cytoskeleton. However, how guidance cues regulate the actin cytoskeleton is not well understood. We report here that UNC-40/DCC, SAX-3/Robo, and VAB-1/Eph differentially regulate the abundance and subcellular localization of the WAVE/SCAR actin nucleation complex and its activator, Rac1/CED-10, in the Caenorhabditis elegans embryonic epidermis. Loss of any of these three pathways results in embryos that fail embryonic morphogenesis. Similar defects in epidermal enclosure have been observed when CED-10/Rac1 or the WAVE/SCAR actin nucleation complex are missing during embryonic development in C. elegans. Genetic and molecular experiments demonstrate that in fact, these three axonal guidance proteins differentially regulate the levels and membrane enrichment of the WAVE/SCAR complex and its activator, Rac1/CED-10, in the epidermis. Live imaging of filamentous actin (F-actin) in embryos developing in the absence of individual guidance receptors shows that high levels of F-actin are not essential for polarized cell migrations, but that properly polarized distribution of F-actin is essential. These results suggest that proper membrane recruitment and activation of CED-10/Rac1 and of WAVE/SCAR by signals at the plasma membrane result in polarized F-actin that permits directed movements and suggest how multiple guidance cues can result in distinct changes in actin nucleation during morphogenesis.
Vyšlo v časopise: UNC-40/DCC, SAX-3/Robo, and VAB-1/Eph Polarize F-Actin during Embryonic Morphogenesis by Regulating the WAVE/SCAR Actin Nucleation Complex. PLoS Genet 8(8): e32767. doi:10.1371/journal.pgen.1002863 Kategorie: Research Article prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002863
1. HedgecockEMCulottiJGHallDH 1990 The unc-5, unc-6, and unc-40 genes guide circumferential migrations of pioneer axons and mesodermal cells on the epidermis of C. elegans. Neuron 4 61 85
2. WadsworthWGBhattHHedgecockEM 1996 Neuroglia and pioneer neurons express UNC-6 to provide global and local netrin cues for guiding migrations in C. elegans. Neuron 16 35 46
3. ZallenJAYiBABargmannCI 1998 The conserved immunoglobulin superfamily member SAX-3/Robo directs multiple aspects of axon guidance in C. elegans. Cell 92 217 27
4. ZallenJAKirchSABargmannCI 1999 Genes required for axon pathfinding and extension in the C. elegans nerve ring. Development 126 3679 92
5. BoulinTPocockRHobertO 2006 A novel Eph receptor-interacting IgSF protein provides C. elegans motorneurons with midline guidepost function. Curr Biol 16 1871 83
6. MohamedAMChin-SangID 2006 Characterization of loss-of-function and gain-of-function Eph receptor tyrosine kinase signaling in C. elegans axon targeting and cell migration. Dev Biol 290 164 76
7. Chin-SangIDGeorgeSEDingMMoseleySLLynchASChisholmAD 1999 The ephrin VAB-2/EFN-1 functions in neuronal signaling to regulate epidermal morphogenesis in C. elegans. Cell 99 781 90
8. GeorgeSESimokatKHardinJChisholmAD 1998 The VAB-1 Eph receptor tyrosine kinase functions in neural and epithelial morphogenesis in C. elegans. Cell 92 633 43
9. GheneaSBoudreauJRLagueNPChin-SangID 2005 The VAB-1 Eph receptor tyrosine kinase and SAX-3/Robo neuronal receptors function together during C. elegans embryonic morphogenesis. Development 132 3679 90
10. HaoJCYuTWFujisawaKCulottiJGGengyo-AndoK 2001 C. elegans slit acts in midline, dorsal-ventral, and anterior-posterior guidance via the SAX-3/Robo receptor. Neuron 32 25 38
11. ChanSSZhengHSuMWWilkRKilleenMT 1996 UNC-40, a C. elegans homolog of DCC (Deleted in Colorectal Cancer), is required in motile cells responding to UNC-6 netrin cues. Cell 87 187 95
12. PatelFPBernadskayaYYChenEJobanputraAPooladiA 2008 The Rac-WAVE/SCAR-Arp2/3 pathway regulates C. elegans morphogenesis by maintaining cytoskeletal and epithelial polarity. Developmental Biology 324 297 309
13. SotoMCQadotaHKasuyaKInoueMTsuboiD 2002 The GEX-2 and GEX-3 proteins are required for tissue morphogenesis and cell migrations in C. elegans. Genes Dev 16 620 32
14. ShakirMAJiangKStruckhoffECDemarcoRSPatelFB 2008 The Arp2/3 activators WAVE and WASP have distinct genetic interactions with Rac GTPases in C. elegans axon guidance. Genetics 179 1957 71
15. BashawGJKleinR 2010 Signaling from axon guidance receptors. Cold Spring Harb Perspect Biol 2 a001941
16. GitaiZYuTWLundquistEATessier-LavigneMBargmannCI 2003 The netrin receptor UNC-40/DCC stimulates axon attraction and outgrowth through enabled and, in parallel, Rac and UNC-115/AbLIM. Neuron 37 53 65
17. LiXSaint-Cyr-ProulxEAktoriesKLamarche-VaneN 2002 Rac1 and Cdc42 but not RhoA or Rho kinase activities are required for neurite outgrowth induced by the Netrin-1 receptor DCC (deleted in colorectal cancer) in N1E-115 neuroblastoma cells. J Biol Chem 277 15207 15214
18. ShekarabiMKennedyTE 2002 The netrin-1 receptor DCC promotes filopodia formation and cell spreading by activating Cdc42 and Rac1. Mol Cell Neurosci 19 1 17
19. ShekarabiMMooreSWTritschNXMorrisSJBouchardJF 2005 Deleted in colorectal cancer binding netrin-1 mediates cell substrate adhesion and recruits Cdc42, Rac1, Pak1, and N-WASP into an intracellular signaling complex that promotes growth cone expansion. J Neurosci 25 3132 3141
20. SahinMGreerPLLinMZPoucherHEberhartJ 2005 Eph-dependent tyrosine phosphorylation of ephexin1 modulates growth cone collapse. Neuron 46 191 204
21. EgeaJNissenUVDufourASahinMGreerP 2005 Regulation of EphA 4 kinase activity is required for a subset of axon guidance decisions suggesting a key role for receptor clustering in Eph function. Neuron 47 515 528
22. FanXLabradorJPHingHBashawGJ 2003 Slit stimulation recruits Dock and Pak to the roundabout receptor and increases Rac activity to regulate axon repulsion at the CNS midline. Neuron 40 113 127
23. Hakeda-SuzukiSNgJTzuJDietzlGSunY 2002 Rac function and regulation during Drosophila development. Nature 416 438 442
24. LundstromAGallioMEnglundCStenebergPHemphalaJ 2004 Vilse, a conserved Rac/Cdc42 GAP mediating Robo repulsion in tracheal cells and axons. Genes Dev 18 2161 2171
25. TotongRAchilleosANanceJ 2007 PAR-6 is required for junction formation but not apicobasal polarization in C. elegans embryonic epithelial cells. Development 134 1259 68
26. Chin-SangIDChisholmAD 2000 Form of the worm: genetics of epidermal morphogenesis in C. elegans. Trends Genet 16 544 51
27. GallyCWisslerFZahreddineHQuintinSLandmannF 2009 Myosin II regulation during C. elegans embryonic elongation: LET-502/ROCK, MRCK-1 and PAK-1, three kinases with different roles. Development 136 3109 19
28. Williams-MassonEMMalikANHardinJ 1997 An actin-mediated two-step mechanism is required for ventral enclosure of the C. elegans hypodermis. Development 124 2889 2901
29. PadrickSBRosenMK 2010 Physical mechanisms of signal integration by WASP family proteins. Annu Rev Biochem 79 707 35
30. KobayashiKKurodaSFukataMNakamuraTNagaseT 1998 p140Sra-1 (specifically Rac1-associated protein) is a novel specific target for Rac1 small GTPase. J Biol Chem 2;273 291 5
31. ChenZBorekDPadrickSBGomezTSMetlagelZ 2010 Structure and control of the actin regulatory WAVE complex. Nature 468 533 8
32. ReddienPWHorvitzHR 2000 CED-2/CrkII and CED-10/Rac control phagocytosis and cell migration in Caenorhabditis elegans. Nat Cell Biol 2 131 6
33. MoorthySChenLBennettV 2000 Caenorhabditis elegans beta-G spectrin is dispensable for establishment of epithelial polarity, but essential for muscular and neuronal function. J Cell Biol 149 915 30
34. BernadskayaYYPatelFBHsuHTSotoMC 2011 Arp2/3 promotes junction formation and maintenance in the C. elegans intestine by regulating membrane association of apical proteins. Molecular Biology of the Cell 22 2886 99
35. LundquistEAReddienPWHartwiegEHorvitzHRBargmannCI 2001 Three C. elegans Rac proteins and several alternative Rac regulators control axon guidance, cell migration and apoptotic cell phagocytosis. Development 128 4475 88
36. BoulterEGarcia-MataRGuilluyCDubashARossiGBrennwaldPJBurridgeK 2010 Regulation of Rho GTPase crosstalk, degradation and activity by RhoGDI1. Nat Cell Biol 12 477 83
37. Garcia-MataRBoulterEBurridgeK 2011 The ‘invisible hand’: regulation of RHO GTPases by RHOGDIs. Nat Rev Mol Cell Biol 12 493 504
38. AndersonP 1995 Mutagenesis. Methods Cell Biol 48 31 58
39. YuTWHaoJCLimWTessier-LavigneMBargmannCI 2002 Shared receptors in axon guidance: SAX-3/Robo signals via UNC-34/Enabled and a Netrin-independent UNC-40/DCC function. Nat Neurosci 5 1147 54
40. MohamedAMBourdreauJRYuFPChin-SangID 2012 The Caenorhabditis elegans Eph receptor activates NCK and N-WASP, and inhibits Ena/VASP to regulate growth cone dynamics during axon guidance. PLoS Genet 2 e1002513 doi:10.1371/journal.pgen.1002513
41. ClarkSGChiuC 2003 C. elegans ZAG-1, a Zn-finger-homeodomain protein, regulates axonal development and neuronal differentiation. Development 130 3781 3794
42. YdenbergCASmithBABreitsprecherDGellesJGoodeBL 2011 Cease-fire at the leading edge: new perspectives on actin filament branching, debranching, and cross-linking. Cytoskeleton 68 596 602
43. BreitsprecherDKoestlerSAChizhovINemethovaMMuellerJGoodeBLSmallJVRottnerKFaixJ 2011 Cofilin cooperates with fascin to disassemble filopodial actin filaments. J Cell Sci 124 3305 18
44. ParsonsJTHorwitzARSchwartzMA 2010 Cell adhesion: integrating cytoskeletal dynamics and cellular tension. Nat Rev Mol Cell Biol 11 633 43
45. PalamidessiAFrittoliEGarréMFarettaMMioneM 2008 Endocytic trafficking of Rac is required for the spatial restriction of signaling in cell migration. Cell 134 135 47
46. BrisbinSLiuJBourdeauJPengJEvangelistaMChin-SangI 2009 A role for C. elegans Eph RTK signaling in PTEN regulation. Dev Cell 17 459 69
47. SuzukiYHanM 2006 Genetic redundancy masks diverse functions of the tumor suppressor gene PTEN during C. elegans development. Genes Dev 20 423 8
48. IkegamiRSimokatKZhengHBrownLGarrigaG 2012 Semaphorin and Eph receptor signaling guide a series of cell movements for ventral enclosure in C. elegans. Curr Biol 22 1 11
49. NoegelAABlau-WasserRSultanaHMüllerRIsraelL 2004 The cyclase-associated protein CAP as regulator of cell polarity and cAMP signaling in Dictyostelium. Mol Biol Cell 15 934 45
50. EllisREJacobsonDMHorvitzHR 1991 Genes required for the engulfment of cell corpses during programmed cell death in Caenorhabditis elegans. Genetics 129 79 94
51. FrancisRWaterstonRH 1991 Muscle cell attachment in Caenorhabditis elegans. J Cell Biol 114 465 479
52. KoppenMSimskeJSSimsPAFiresteinBLHallDH 2001 Cooperative regulation of AJM-1 controls junctional integrity in Caenorhabditis elegans epithelia. Nat Cell Biol 3 983 991
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