Tiam–Rac signaling mediates trans-endocytosis of ephrin receptor EphB2 and is important for cell repulsion

Journal of Cell Biology - Tập 214 Số 6 - Trang 735-752 - 2016
Thomas N. Gaitanos1, Jorg Koerner1, Rüdiger Klein1
1Department of Molecules-Signaling-Development, Max Planck Institute of Neurobiology, 82152 Munich-Martinsried, Germany

Tóm tắt

Ephrin receptors interact with membrane-bound ephrin ligands to regulate contact-mediated attraction or repulsion between opposing cells, thereby influencing tissue morphogenesis. Cell repulsion requires bidirectional trans-endocytosis of clustered Eph–ephrin complexes at cell interfaces, but the mechanisms underlying this process are poorly understood. Here, we identified an actin-regulating pathway allowing ephrinB+ cells to trans-endocytose EphB receptors from opposing cells. Live imaging revealed Rac-dependent F-actin enrichment at sites of EphB2 internalization, but not during vesicle trafficking. Systematic depletion of Rho family GTPases and their regulatory proteins identified the Rac subfamily and the Rac-specific guanine nucleotide exchange factor Tiam2 as key components of EphB2 trans-endocytosis, a pathway previously implicated in Eph forward signaling, in which ephrins act as in trans ligands of Eph receptors. However, unlike in Eph signaling, this pathway is not required for uptake of soluble ligands in ephrinB+ cells. We also show that this pathway is required for EphB2-stimulated contact repulsion. These results support the existence of a conserved pathway for EphB trans-endocytosis that removes the physical tether between cells, thereby enabling cell repulsion.

Từ khóa


Tài liệu tham khảo

Batlle, 2012, Molecular mechanisms of cell segregation and boundary formation in development and tumorigenesis, Cold Spring Harb. Perspect. Biol., 4, a008227, 10.1101/cshperspect.a008227

Boissier, 2013, EphA2 signaling following endocytosis: role of Tiam1, Traffic., 14, 1255, 10.1111/tra.12123

Carpenter, 2006, CellProfiler: image analysis software for identifying and quantifying cell phenotypes, Genome Biol., 7, R100, 10.1186/gb-2006-7-10-r100

Cook, 2014, Rho guanine nucleotide exchange factors: regulators of Rho GTPase activity in development and disease, Oncogene., 33, 4021, 10.1038/onc.2013.362

Corbetta, 2009, Essential role of Rac1 and Rac3 GTPases in neuronal development, FASEB J., 23, 1347, 10.1096/fj.08-121574

Cowan, 2005, Vav family GEFs link activated Ephs to endocytosis and axon guidance, Neuron., 46, 205, 10.1016/j.neuron.2005.03.019

Csépányi-Kömi, 2012, ARHGAP25, a novel Rac GTPase-activating protein, regulates phagocytosis in human neutrophilic granulocytes, Blood., 119, 573, 10.1182/blood-2010-12-324053

Davis, 1994, Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity, Science., 266, 816, 10.1126/science.7973638

Deininger, 2008, The Rab5 guanylate exchange factor Rin1 regulates endocytosis of the EphA4 receptor in mature excitatory neurons, Proc. Natl. Acad. Sci. USA., 105, 12539, 10.1073/pnas.0801174105

Dyer, 2010, Distinct roles of Rac GTPases and the UNC-73/Trio and PIX-1 Rac GTP exchange factors in neuroblast protrusion and migration in C. elegans, Small GTPases., 1, 44, 10.4161/sgtp.1.1.12991

Franke, 2012, miR-124-regulated RhoG reduces neuronal process complexity via ELMO/Dock180/Rac1 and Cdc42 signalling, EMBO J., 31, 2908, 10.1038/emboj.2012.130

Gaitanos, 2004, Peloruside A does not bind to the taxoid site on beta-tubulin and retains its activity in multidrug-resistant cell lines, Cancer Res., 64, 5063, 10.1158/0008-5472.CAN-04-0771

Gong, 2016, Exosomes mediate cell contact-independent ephrin-Eph signaling during axon guidance, J. Cell Biol., 214, 35, 10.1083/jcb.201601085

Groeger, 2007, Co-operative Cdc42 and Rho signalling mediates ephrinB-triggered endothelial cell retraction, Biochem. J., 404, 23, 10.1042/BJ20070146

Gu, 2003, Hematopoietic cell regulation by Rac1 and Rac2 guanosine triphosphatases, Science., 302, 445, 10.1126/science.1088485

Heasman, 2008, Mammalian Rho GTPases: new insights into their functions from in vivo studies, Nat. Rev. Mol. Cell Biol., 9, 690, 10.1038/nrm2476

Himanen, 2010, Architecture of Eph receptor clusters, Proc. Natl. Acad. Sci. USA., 107, 10860, 10.1073/pnas.1004148107

Huber, 2008, FGD2, a CDC42-specific exchange factor expressed by antigen-presenting cells, localizes to early endosomes and active membrane ruffles, J. Biol. Chem., 283, 34002, 10.1074/jbc.M803957200

Irie, 2002, EphB receptors regulate dendritic spine development via intersectin, Cdc42 and N-WASP, Nat. Neurosci., 5, 1117, 10.1038/nn964

Itoh, 2002, Activation of rac and cdc42 video imaged by fluorescent resonance energy transfer-based single-molecule probes in the membrane of living cells, Mol. Cell. Biol., 22, 6582, 10.1128/MCB.22.18.6582-6591.2002

Jaiswal, 2013, Deciphering the molecular and functional basis of Dbl family proteins: a novel systematic approach toward classification of selective activation of the Rho family proteins, J. Biol. Chem., 288, 4486, 10.1074/jbc.M112.429746

Jørgensen, 2009, Cell-specific information processing in segregating populations of Eph receptor ephrin-expressing cells, Science., 326, 1502, 10.1126/science.1176615

Joset, 2010, Pincher-generated Nogo-A endosomes mediate growth cone collapse and retrograde signaling, J. Cell Biol., 188, 271, 10.1083/jcb.200906089

Kaneko, 2005, Rho mediates endocytosis of epidermal growth factor receptor through phosphorylation of endophilin A1 by Rho-kinase, Genes Cells., 10, 973, 10.1111/j.1365-2443.2005.00895.x

Kania, 2016, Mechanisms of ephrin-Eph signalling in development, physiology and disease, Nat. Rev. Mol. Cell Biol., 17, 240, 10.1038/nrm.2015.16

Khelfaoui, 2009, Inhibition of RhoA pathway rescues the endocytosis defects in Oligophrenin1 mouse model of mental retardation, Hum. Mol. Genet., 18, 2575, 10.1093/hmg/ddp189

Klein, 2012, Eph/ephrin signalling during development, Development., 139, 4105, 10.1242/dev.074997

Kullander, 2001, Ephrin-B3 is the midline barrier that prevents corticospinal tract axons from recrossing, allowing for unilateral motor control, Genes Dev., 15, 877, 10.1101/gad.868901

Lauterbach, 2006, Release of full-length EphB2 receptors from hippocampal neurons to cocultured glial cells, J. Neurosci., 26, 11575, 10.1523/JNEUROSCI.2697-06.2006

Lisabeth, 2013, Eph receptor signaling and ephrins, Cold Spring Harb. Perspect. Biol., 5, a009159, 10.1101/cshperspect.a009159

Marchesi, 2014, DEPDC1B coordinates de-adhesion events and cell-cycle progression at mitosis, Dev. Cell., 31, 420, 10.1016/j.devcel.2014.09.009

Marston, 2003, Rac-dependent trans-endocytosis of ephrinBs regulates Eph-ephrin contact repulsion, Nat. Cell Biol., 5, 879, 10.1038/ncb1044

Nakano-Kobayashi, 2009, The Rho-linked mental retardation protein OPHN1 controls synaptic vesicle endocytosis via endophilin A1, Curr. Biol., 19, 1133, 10.1016/j.cub.2009.05.022

Nakaya, 2006, Opposite effects of rho family GTPases on engulfment of apoptotic cells by macrophages, J. Biol. Chem., 281, 8836, 10.1074/jbc.M510972200

Negrete, 2005, EphrinB2 is the entry receptor for Nipah virus, an emergent deadly paramyxovirus, Nature., 436, 401, 10.1038/nature03838

Nishimura, 2006, Role of numb in dendritic spine development with a Cdc42 GEF intersectin and EphB2, Mol. Biol. Cell., 17, 1273, 10.1091/mbc.E05-07-0700

Parker, 2004, Reverse endocytosis of transmembrane ephrin-B ligands via a clathrin-mediated pathway, Biochem. Biophys. Res. Commun., 323, 17, 10.1016/j.bbrc.2004.07.209

Pasquale, 2008, Eph-ephrin bidirectional signaling in physiology and disease, Cell., 133, 38, 10.1016/j.cell.2008.03.011

Pasquale, 2010, Eph receptors and ephrins in cancer: bidirectional signalling and beyond, Nat. Rev. Cancer., 10, 165, 10.1038/nrc2806

Pernet, 2009, Nipah virus entry can occur by macropinocytosis, Virology., 395, 298, 10.1016/j.virol.2009.09.016

Qin, 2010, Structural characterization of the EphA4-Ephrin-B2 complex reveals new features enabling Eph-ephrin binding promiscuity, J. Biol. Chem., 285, 644, 10.1074/jbc.M109.064824

Riedl, 2008, Lifeact: a versatile marker to visualize F-actin, Nat. Methods., 5, 605, 10.1038/nmeth.1220

Roberts, 2008, Rho Family GTPase modification and dependence on CAAX motif-signaled posttranslational modification, J. Biol. Chem., 283, 25150, 10.1074/jbc.M800882200

Rooney, 2010, The Rac activator STEF (Tiam2) regulates cell migration by microtubule-mediated focal adhesion disassembly, EMBO Rep., 11, 292, 10.1038/embor.2010.10

Sahin, 2005, Eph-dependent tyrosine phosphorylation of ephexin1 modulates growth cone collapse, Neuron., 46, 191, 10.1016/j.neuron.2005.01.030

Schaupp, 2014, The composition of EphB2 clusters determines the strength in the cellular repulsion response, J. Cell Biol., 204, 409, 10.1083/jcb.201305037

Schindelin, 2012, Fiji: an open-source platform for biological-image analysis, Nat. Methods., 9, 676, 10.1038/nmeth.2019

Seiradake, 2010, An extracellular steric seeding mechanism for Eph-ephrin signaling platform assembly, Nat. Struct. Mol. Biol., 17, 398, 10.1038/nsmb.1782

Seiradake, 2014, FLRT structure: balancing repulsion and cell adhesion in cortical and vascular development, Neuron., 84, 370, 10.1016/j.neuron.2014.10.008

Shamah, 2001, EphA receptors regulate growth cone dynamics through the novel guanine nucleotide exchange factor ephexin, Cell., 105, 233, 10.1016/S0092-8674(01)00314-2

Shutes, 2007, Specificity and mechanism of action of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases, J. Biol. Chem., 282, 35666, 10.1074/jbc.M703571200

Smyth, 2012, Actin cytoskeleton rest stops regulate anterograde traffic of connexin 43 vesicles to the plasma membrane, Circ. Res., 110, 978, 10.1161/CIRCRESAHA.111.257964

Swanson, 2008, Shaping cups into phagosomes and macropinosomes, Nat. Rev. Mol. Cell Biol., 9, 639, 10.1038/nrm2447

Takeuchi, 2015, Eph/ephrin reverse signalling induces axonal retraction through RhoA/ROCK pathway, J. Biochem., 158, 245, 10.1093/jb/mvv042

Tanaka, 2004, Tiam1 mediates neurite outgrowth induced by ephrin-B1 and EphA2, EMBO J., 23, 1075, 10.1038/sj.emboj.7600128

Tcherkezian, 2007, Current knowledge of the large RhoGAP family of proteins, Biol. Cell., 99, 67, 10.1042/BC20060086

Terawaki, 2010, The PHCCEx domain of Tiam1/2 is a novel protein- and membrane-binding module, EMBO J., 29, 236, 10.1038/emboj.2009.323

Tolias, 2007, The Rac1 guanine nucleotide exchange factor Tiam1 mediates EphB receptor-dependent dendritic spine development, Proc. Natl. Acad. Sci. USA., 104, 7265, 10.1073/pnas.0702044104

Um, 2014, Dynamic control of excitatory synapse development by a Rac1 GEF/GAP regulatory complex, Dev. Cell., 29, 701, 10.1016/j.devcel.2014.05.011

Wennerberg, 2002, RhoG signals in parallel with Rac1 and Cdc42, J. Biol. Chem., 277, 47810, 10.1074/jbc.M203816200

Xu, 2009, Ephrin-B3 reverse signaling through Grb4 and cytoskeletal regulators mediates axon pruning, Nat. Neurosci., 12, 268, 10.1038/nn.2254

Xu, 2013, Boundary formation in the development of the vertebrate hindbrain, Wiley Interdiscip. Rev. Dev. Biol., 2, 735, 10.1002/wdev.106

Yoo, 2010, EphA8-ephrinA5 signaling and clathrin-mediated endocytosis is regulated by Tiam-1, a Rac-specific guanine nucleotide exchange factor, Mol. Cells., 29, 603, 10.1007/s10059-010-0075-2

Zhuang, 2007, Regulation of EphA2 receptor endocytosis by SHIP2 lipid phosphatase via phosphatidylinositol 3-Kinase-dependent Rac1 activation, J. Biol. Chem., 282, 2683, 10.1074/jbc.M608509200

Zimmer, 2003, EphB-ephrinB bi-directional endocytosis terminates adhesion allowing contact mediated repulsion, Nat. Cell Biol., 5, 869, 10.1038/ncb1045