Bipartite Inhibition of Drosophila Epidermal Growth Factor Receptor by the Extracellular and Transmembrane Domains of Kekkon1

Genetics - Tập 167 Số 1 - Trang 187-202 - 2004
Diego Alvarado1, Amy H Rice2, Joseph B. Duffy2
1Department of Biology, Indiana University, Bloomington, Indiana 47405, USA
2Department of Biology, Indiana University, Bloomington, Indiana 47405

Tóm tắt

AbstractIn Drosophila, signaling by the epidermal growth factor receptor (EGFR) is required for a diverse array of developmental decisions. Essential to these decisions is the precise regulation of the receptor's activity by both stimulatory and inhibitory molecules. To better understand the regulation of EGFR activity we investigated inhibition of EGFR by the transmembrane protein Kekkon1 (Kek1). Kek1 encodes a molecule containing leucine-rich repeats (LRR) and an immunoglobulin (Ig) domain and is the founding member of the Drosophila Kekkon family. Here we demonstrate with a series of Kek1-Kek2 chimeras that while the LRRs suffice for EGFR binding, inhibition in vivo requires the Kek1 juxta/transmembrane region. We demonstrate directly, and using a series of Kek1-EGFR chimeras, that Kek1 is not a phosphorylation substrate for the receptor in vivo. In addition, we show that EGFR inhibition is unique to Kek1 among Kek family members and that this function is not ligand or tissue specific. Finally, we have identified a unique class of EGFR alleles that specifically disrupt Kek1 binding and inhibition, but preserve receptor activation. Interestingly, these alleles map to domain V of the Drosophila EGFR, a region absent from the vertebrate receptors. Together, our results support a model in which the LRRs of Kek1 in conjunction with its juxta/transmembrane region direct association and inhibition of the Drosophila EGFR through interactions with receptor domain V.

Từ khóa


Tài liệu tham khảo

Adams, 2000, The genome sequence of Drosophila melanogaster, Science, 287, 2185, 10.1126/science.287.5461.2185

Alroy, 1997, The ErbB signaling network in embryogenesis and oncogenesis: signal diversification through combinatorial ligand-receptor interactions, FEBS Lett., 410, 83, 10.1016/S0014-5793(97)00412-2

Alvarado, 2004, Knockouts of Kekkon1 define sequence elements essential for Drosophila epidermal growth factor receptor inhibition, Genetics, 166, 201, 10.1534/genetics.166.1.201

Aroian, 1990, The let-23 gene necessary for C. elegans vulval induction encodes a tyrosine kinase of the EGF receptor sub-family, Nature, 348, 693, 10.1038/348693a0

Ashburner, 1989, Drosophila: A Laboratory Handbook

Blume-Jensen, 2001, Oncogenic kinase signalling, Nature, 411, 355, 10.1038/35077225

Boutros, 2000, Signaling specificity by Frizzled receptors in Drosophila., Science, 288, 1825, 10.1126/science.288.5472.1825

Burgess, 2003, An open-and-shut case? Recent insights into the activation of the EGF/ErbB receptors, Mol. Cell, 12, 541, 10.1016/S1097-2765(03)00350-2

Casci, 1999, Sprouty, an intracellular inhibitor of Ras signaling, Cell, 96, 655, 10.1016/S0092-8674(00)80576-0

Cherbas, 1998, Cell culture, Drosophila: A Practical Approach, 319, 10.1093/oso/9780199636617.003.0010

Clifford, 1989, Coordinately and differentially mutable activities of torpedo, the Drosophila melanogaster homologue of the vertebrate EGF receptor gene, Genetics, 123, 771, 10.1093/genetics/123.4.771

Clifford, 1994, Molecular analysis of the Drosophila EGF receptor homolog reveals that several genetically defined classes of alleles cluster in subdomains of the receptor protein, Genetics, 137, 531, 10.1093/genetics/137.2.531

Csordas, 2000, Sustained down-regulation of the epidermal growth factor receptor by decorin. A mechanism for controlling tumor growth in vivo., J. Biol. Chem., 275, 32879, 10.1074/jbc.M005609200

Derheimer, 2004, Conservation of an inhibitor of the epidermal growth factor receptor, Kekkon1, in dipterans, Genetics, 166, 213, 10.1534/genetics.166.1.213

Diaz-Benjumea, 1990, Behaviour of cells mutant for an EGF receptor homologue of Drosophila in genetic mosaics, Proc. R. Soc. Lond. Ser. B Biol. Sci., 242, 36, 10.1098/rspb.1990.0100

Dickson, 1992, Prepattern in the developing Drosophila eye revealed by an activated torso-sevenless chimeric receptor, Genes Dev., 6, 2327, 10.1101/gad.6.12a.2327

Dominguez, 1998, Multiple functions of the EGF receptor in Drosophila eye development, Curr. Biol., 8, 1039, 10.1016/S0960-9822(98)70441-5

Fehon, 1990, Molecular interactions between the protein products of the neurogenic loci Notch and Delta, two EGF-homologous genes in Drosophila., Cell, 61, 523, 10.1016/0092-8674(90)90534-L

Ferguson, 2003, EGF activates its receptor by removing interactions that autoinhibit ectodomain dimerization, Mol. Cell, 11, 507, 10.1016/S1097-2765(03)00047-9

Freeman, 1996, Reiterative use of the EGF receptor triggers differentiation of all cell types in the Drosophila eye, Cell, 87, 651, 10.1016/S0092-8674(00)81385-9

Freeman, 1998, Complexity of EGF receptor signalling revealed in Drosophila., Curr. Opin. Genet. Dev., 8, 407, 10.1016/S0959-437X(98)80110-X

Garrett, 2002, Crystal structure of a truncated epidermal growth factor receptor extracellular domain bound to transforming growth factor alpha, Cell, 110, 763, 10.1016/S0092-8674(02)00940-6

Garrett, 2003, The crystal structure of a truncated ErbB2 ectodomain reveals an active conformation, poised to interact with other ErbB receptors, Mol. Cell, 11, 495, 10.1016/S1097-2765(03)00048-0

Ghiglione, 1999, The transmembrane molecule kekkon 1 acts in a feedback loop to negatively regulate the activity of the Drosophila EGF receptor during oogenesis, Cell, 96, 847, 10.1016/S0092-8674(00)80594-2

Ghiglione, 2003, Mechanism of inhibition of the Drosophila and mammalian EGF receptors by the transmembrane protein Kekkon 1, Development, 130, 4483, 10.1242/dev.00617

Gotoh, 1992, A highly conserved tyrosine residue at codon 845 within the kinase domain is not required for the transforming activity of human epidermal growth factor receptor, Biochem. Biophys. Res. Commun., 186, 768, 10.1016/0006-291X(92)90812-Y

Guichard, 1999, rhomboid and Star interact synergistically to promote EGFR/MAPK signaling during Drosophila wing vein development, Development, 126, 2663, 10.1242/dev.126.12.2663

Guichard, 2002, A screen for dominant mutations applied to components in the Drosophila EGF-R pathway, Proc. Natl. Acad. Sci. USA, 99, 3752, 10.1073/pnas.052028699

Hanks, 1988, The protein kinase family: conserved features and deduced phyologeny of the catalytic domains, Science, 241, 42, 10.1126/science.3291115

Harari, 2000, Molecular mechanisms underlying ErbB2/HER2 action in breast cancer, Oncogene, 19, 6102, 10.1038/sj.onc.1203973

Hime, 1997, D-Cbl, the Drosophila homologue of the c-Cbl proto-oncogene, interacts with the Drosophila EGF receptor in vivo, despite lacking C-terminal adaptor binding sites, Oncogene, 14, 2709, 10.1038/sj.onc.1201223

Holbro, 2003, The ErbB receptors and their role in cancer progression, Exp. Cell Res., 284, 99, 10.1016/S0014-4827(02)00099-X

Hubbard, 2000, Protein tyrosine kinase structure and function, Annu. Rev. Biochem., 69, 373, 10.1146/annurev.biochem.69.1.373

Huppert, 1997, Feedback regulation is central to Delta-Notch signalling required for Drosophila wing vein morphogenesis, Development, 124, 3283, 10.1242/dev.124.17.3283

Iozzo, 1999, Decorin is a biological ligand for the epidermal growth factor receptor, J. Biol. Chem., 274, 4489, 10.1074/jbc.274.8.4489

Keleman, 2001, Short- and long-range repulsion by the Drosophila Unc5 netrin receptor, Neuron, 32, 605, 10.1016/S0896-6273(01)00505-0

Keleman, 2002, Comm sorts robo to control axon guidance at the Drosophila midline, Cell, 110, 415, 10.1016/S0092-8674(02)00901-7

Kim, 1998, Signal transduction by epidermal growth factor and heregulin via the kinase-deficient ErbB3 protein, Biochem. J., 334, 189, 10.1042/bj3340189

Kimmel, 1990, The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype, Genes Dev., 4, 712, 10.1101/gad.4.5.712

Klingbeil, 1999, A basic residue, Lys 782, composes part of the ATP-binding site on the epidermal growth factor receptor tyrosine kinase, Arch. Biochem. Biophys., 363, 27, 10.1006/abbi.1998.1052

Klueg, 2002, Creation of a GAL4/UAS-coupled inducible gene expression system for use in Drosophila cultured cell lines, Genesis, 34, 119, 10.1002/gene.10148

Kramer, 1999, Sprouty: a common antagonist of FGF and EGF signaling pathways in Drosophila., Development, 126, 2515, 10.1242/dev.126.11.2515

Kuja-Panula, 2003, AMIGO, a transmembrane protein implicated in axon tract development, defines a novel protein family with leucine-rich repeats, J. Cell Biol., 160, 963, 10.1083/jcb.200209074

Lax, 1988, Localization of a major receptor-binding domain for epidermal growth factor by affinity labeling, Mol. Cell Biol., 8, 1831

Lee, 1995, Requirement for neuregulin receptor erbB2 in neural and cardiac development, Nature, 378, 394, 10.1038/378394a0

Lee, 1999, Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis, Neuron, 22, 451, 10.1016/S0896-6273(00)80701-1

Lesokhin, 1999, Several levels of EGF receptor signaling during photoreceptor specification in wild-type, Ellipse, and null mutant Drosophila., Dev. Biol., 205, 129, 10.1006/dbio.1998.9121

Moriki, 2001, Activation of preformed EGF receptor dimers by ligand-induced rotation of the transmembrane domain, J. Mol. Biol., 311, 1011, 10.1006/jmbi.2001.4923

Murphy, 1995, The breathless FGF receptor homolog, a downstream target of Drosophila C/EBP in the developmental control of cell migration, Development, 121, 2255, 10.1242/dev.121.8.2255

Musacchio, 1996, The Drosophila kekkon genes: novel members of both the leucine-rich repeat and immunoglobulin superfamilies expressed in the CNS, Dev. Biol., 178, 63, 10.1006/dbio.1996.0198

Neuman-Silberberg, 1993, The Drosophila dorsoventral patterning gene gurken produces a dorsally localized RNA and encodes a TGFα-like protein, Cell, 75, 165, 10.1016/S0092-8674(05)80093-5

Nilson, 1999, EGF receptor signaling in Drosophila oogenesis, Curr. Top. Dev. Biol., 44, 203, 10.1016/S0070-2153(08)60471-8

Ogiso, 2002, Crystal structure of the complex of human epidermal growth factor and receptor extracellular domains, Cell, 110, 775, 10.1016/S0092-8674(02)00963-7

Olayioye, 2000, The ErbB signaling network: receptor heterodimerization in development and cancer, EMBO J., 19, 3159, 10.1093/emboj/19.13.3159

Pai, 2000, D-cbl, a negative regulator of the Egfr pathway, is required for dorsoventral patterning in Drosophila oogenesis, Cell, 103, 51, 10.1016/S0092-8674(00)00104-5

Patel, 1998, Decorin activates the epidermal growth factor receptor and elevates cytosolic Ca2+ in A431 carcinoma cells, J. Biol. Chem., 273, 3121, 10.1074/jbc.273.6.3121

Price, 1989, The maternal ventralizing locus torpedo is allelic to faint little ball, an embryonic lethal, and encodes the Drosophila EGF receptor homolog, Cell, 56, 1085, 10.1016/0092-8674(89)90641-7

Qian, 1995, Intermolecular association and trans-phosphorylation of different neu-kinase forms permit SH2-dependent signaling and oncogenic transformation, Oncogene, 10, 211

Queenan, 1997, Ectopic activation of torpedo/Egfr, a Drosophila receptor tyrosine kinase, dorsalizes both the eggshell and the embryo, Development, 124, 3871, 10.1242/dev.124.19.3871

Raz, 1991, Interallelic complementation among DER/flb alleles: implications for the mechanism of signal transduction by receptor-tyrosine kinases, Genetics, 129, 191, 10.1093/genetics/129.1.191

Reichman-Fried, 1994, Elucidation of the role of breathless, a Drosophila FGF receptor homolog, in tracheal cell migration, Genes Dev., 8, 428, 10.1101/gad.8.4.428

Roth, 1995, cornichon and the EGF receptor signaling process are necessary for both anterior-posterior and dorsal-ventral pattern formation in Drosophila., Cell, 81, 967, 10.1016/0092-8674(95)90016-0

Santra, 2000, An anti-oncogenic role for decorin. Down-regulation of ErbB2 leads to growth suppression and cytodifferentiation of mammary carcinoma cells, J. Biol. Chem., 275, 35153, 10.1074/jbc.M006821200

Santra, 2002, Decorin binds to a narrow region of the epidermal growth factor (EGF) receptor, partially overlapping but distinct from the EGF-binding epitope, J. Biol. Chem., 277, 35671, 10.1074/jbc.M205317200

Schlessinger, 2000, Cell signaling by receptor tyrosine kinases, Cell, 103, 211, 10.1016/S0092-8674(00)00114-8

Schlessinger, 2002, All signaling is local?, Mol. Cell, 10, 218, 10.1016/S1097-2765(02)00607-X

Schlessinger, 2003, SH2 and PTB domains in tyrosine kinase signaling, Sci. STKE 2003: RE12.

Schnepp, 1998, EGF domain swap converts a Drosophila EGF receptor activator into an inhibitor, Genes Dev., 12, 908, 10.1101/gad.12.7.908

Schweitzer, 1997, A thousand and one roles for the Drosophila EGF receptor, Trends Genet., 13, 191, 10.1016/S0168-9525(97)01091-3

Schweitzer, 1995, Inhibition of Drosophila EGF receptor activation by the secreted protein Argos, Nature, 376, 699, 10.1038/376699a0

Simcox, 1997, Differential requirement for EGF-like ligands in Drosophila wing development, Mech. Dev., 62, 41, 10.1016/S0925-4773(96)00643-0

Sliwkowski, 1994, Coexpression of erbB2 and erbB3 proteins reconstitutes a high affinity receptor for heregulin, J. Biol. Chem., 269, 14661, 10.1016/S0021-9258(17)36676-0

Speicher, 1998, Neurotactin functions in concert with other identified CAMs in growth cone guidance in Drosophila., Neuron, 20, 221, 10.1016/S0896-6273(00)80451-1

Stamos, 2002, Structure of the EGF receptor kinase domain alone and in complex with a 4-anilinoquinazoline inhibitor, J. Biol. Chem., 277, 46265, 10.1074/jbc.M207135200

Stevens, 1998, Twin peaks: Spitz and Argos star in patterning of the Drosophila egg, Cell, 95, 291, 10.1016/S0092-8674(00)81759-6

Stoscheck, 1986, Role of epidermal growth factor in carcinogenesis, Cancer Res., 46, 1030

Sturtevant, 1993, The Drosophila rhomboid gene mediates the localized formation of wing veins and interacts genetically with components of the EGF-R signaling pathway, Genes Dev., 7, 961, 10.1101/gad.7.6.961

Tio, 1997, The Drosophila TGF alpha homolog Spitz acts in photoreceptor recruitment in the developing retina, Development, 124, 343, 10.1242/dev.124.2.343

Ullrich, 1990, Signal transduction by receptors with tyrosine kinase activity, Cell, 61, 203, 10.1016/0092-8674(90)90801-K

Van Buskirk, 1999, Versatility in signalling: multiple responses to EGF receptor activation during Drosophila oogenesis, Trends Cell Biol., 9, 1, 10.1016/S0962-8924(98)01413-5

Wasserman, 1998, An autoregulatory cascade of EGF receptor signaling patterns the Drosophila egg, Cell, 95, 355, 10.1016/S0092-8674(00)81767-5

Wieschaus, 1984, Mutations affecting the pattern of the larval cuticle in Drosophila melanogaster. 3. Zygotic loci on the X-chromosome and the 4th chromosome. Wilhelm Roux’s Arch, Dev. Biol., 193, 296

Yarden, 1987, Epidermal growth factor induces rapid, reversible aggregation of the purified epidermal growth factor receptor, Biochemistry, 26, 1443, 10.1021/bi00379a035

Yarden, 2001, Untangling the ErbB signalling network, Nat. Rev. Mol. Cell. Biol., 2, 127, 10.1038/35052073

Yarden, 1988, Growth factor receptor tyrosine kinases, Annu. Rev. Biochem., 57, 443, 10.1146/annurev.bi.57.070188.002303