Mammalian NOTCH-1 Activates β1 Integrins via the Small GTPase R-Ras*

Notch is a central regulator of important cell fate decisions. Notch activation produces diverse cellular effects suggesting the presence of context-dependent control mechanisms. Genetic studies have demonstrated that Notch and integrin mutations have related phenotypes in key developmental processes such as vascular development and somitogenesis. We show that the intracellular domain of mammalian Notch-1 activates integrins without affecting integrin expression. Integrin activation is dependent on γ-secretase-mediated intramembranous cleavage of membrane-bound Notch releasing intracellular Notch that activates R-Ras, independent of CSL-transcription. Notch also reverses H-Ras and Raf-mediated integrin suppression without affecting ERK phosphorylation. Membrane-bound Notch mutants that are inefficiently cleaved or intracellular Notch mutants lacking the ankyrin repeat sequence do not activate R-Ras or integrins. Co-expression of Msx2-interacting nuclear target (MINT) protein with Notch or expression of intracellular Notch-1 truncation mutants lacking the C-terminal transactivation/PEST domain suppresses Notch transcriptional activity without affecting integrin activation. Notch ligand, Delta-like ligand-4, stimulates R-Ras-dependent α5β1 integrin-mediated adhesion, demonstrating the physiological relevance of this pathway. This new CSL-independent Notch/R-Ras pathway provides a molecular mechanism to explain Notch, integrin, and Ras cross-talk during the development of multicellular organisms.

[1]  C. Haslett,et al.  Phospholipase Cϵ Suppresses Integrin Activation* , 2006, Journal of Biological Chemistry.

[2]  A. Friday,et al.  Notch receptor encodes two structurally separable functions in Drosophila: A genetic analysis , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[3]  Lia S. Campos,et al.  Notch, Epidermal Growth Factor Receptor, and β1-Integrin Pathways Are Coordinated in Neural Stem Cells* , 2006, Journal of Biological Chemistry.

[4]  E. Ruoslahti,et al.  R-Ras is a global regulator of vascular regeneration that suppresses intimal hyperplasia and tumor angiogenesis , 2005, Nature Medicine.

[5]  Shang‐Wei Chong,et al.  Off limits--integrins holding boundaries in somitogenesis. , 2005, Trends in cell biology.

[6]  M. Sundaram,et al.  The love-hate relationship between Ras and Notch. , 2005, Genes & development.

[7]  T. Shimazaki,et al.  Prospective characterization of neural stem cells by flow cytometry analysis using a combination of surface markers , 2005, Journal of neuroscience research.

[8]  R. Geisler,et al.  Integrinalpha5 and delta/notch signaling have complementary spatiotemporal requirements during zebrafish somitogenesis. , 2005, Developmental cell.

[9]  U. Banerjee,et al.  The little R cell that could. , 2004, International Journal of Developmental Biology.

[10]  Andrés J. García,et al.  Specific β1 integrins mediate adhesion, migration, and differentiation of neural progenitors derived from the embryonic striatum , 2004, Molecular and Cellular Neuroscience.

[11]  M. Sundaram,et al.  Vulval Development: The Battle between Ras and Notch , 2004, Current Biology.

[12]  M. Wolfe,et al.  Notch 1 signaling regulates peripheral T cell activation. , 2004, Immunity.

[13]  Raphael Kopan,et al.  Anchoring notch genetics and biochemistry; structural analysis of the ankyrin domain sheds light on existing data. , 2004, Molecular cell.

[14]  W. Pear,et al.  Notch and the immune system. , 2003, Immunity.

[15]  K. Raj,et al.  The role of Notch in tumorigenesis: oncogene or tumour suppressor? , 2003, Nature Reviews Cancer.

[16]  J. Furuhjelm,et al.  The C-terminal end of R-Ras contains a focal adhesion targeting signal , 2003, Journal of Cell Science.

[17]  D. Clegg,et al.  Integrins in the development, function and dysfunction of the nervous system. , 2003, Frontiers in bioscience : a journal and virtual library.

[18]  Larry Kedes,et al.  HES and HERP families: Multiple effectors of the notch signaling pathway , 2003, Journal of cellular physiology.

[19]  J. Alberola-Ila,et al.  The Ras/MAPK cascade and the control of positive selection , 2003, Immunological reviews.

[20]  T. Honjo,et al.  Regulation of marginal zone B cell development by MINT, a suppressor of Notch/RBP-J signaling pathway. , 2003, Immunity.

[21]  Yan Li,et al.  Regulation of Notch1 and Dll4 by Vascular Endothelial Growth Factor in Arterial Endothelial Cells: Implications for Modulating Arteriogenesis and Angiogenesis , 2003, Molecular and Cellular Biology.

[22]  J. Schrader,et al.  Ras and relatives--job sharing and networking keep an old family together. , 2002, Experimental hematology.

[23]  Keith Brennan,et al.  CSL-independent Notch signalling: a checkpoint in cell fate decisions during development? , 2002, Current opinion in genetics & development.

[24]  R. Hynes.  A reevaluation of integrins as regulators of angiogenesis , 2002, Nature Medicine.

[25]  M. Hansen,et al.  Suppression of integrin activation by activated Ras or Raf does not correlate with bulk activation of ERK MAP kinase. , 2002, Molecular biology of the cell.

[26]  U. Suter,et al.  Notch1 and its ligands Delta-like and Jagged are expressed and active in distinct cell populations in the postnatal mouse brain , 2002, Mechanisms of Development.

[27]  L. Hood,et al.  Activated Notch4 Inhibits Angiogenesis: Role of β1-Integrin Activation , 2002, Molecular and Cellular Biology.

[28]  Yukinori Endo,et al.  Bimodal functions of Notch-mediated signaling are involved in neural crest formation during avian ectoderm development. , 2002, Development.

[29]  P. Heitzler,et al.  Novel Notch alleles reveal a Deltex-dependent pathway repressing neural fate , 2001, Current Biology.

[30]  E. Formstecher,et al.  PEA-15 mediates cytoplasmic sequestration of ERK MAP kinase. , 2001, Developmental cell.

[31]  S. Baylin,et al.  Notch signaling induces cell cycle arrest in small cell lung cancer cells. , 2001, Cancer research.

[32]  A. Hajnal,et al.  Notch Inhibition of RAS Signaling Through MAP Kinase Phosphatase LIP-1 During C. elegans Vulval Development , 2001, Science.

[33]  J. Modolell,et al.  The EGF receptor and N signalling pathways act antagonistically in Drosophila mesothorax bristle patterning. , 2001, Development.

[34]  G. Weinmaster,et al.  Ligand-induced signaling in the absence of furin processing of Notch1. , 2001, Developmental biology.

[35]  I. Bernstein,et al.  Immobilization of Notch ligand, Delta-1, is required for induction of notch signaling. , 2000, Journal of cell science.

[36]  Jon C. Aster,et al.  Essential Roles for Ankyrin Repeat and Transactivation Domains in Induction of T-Cell Leukemia by Notch1 , 2000, Molecular and Cellular Biology.

[37]  J. Downward,et al.  The effector loop and prenylation site of R-Ras are involved in the regulation of integrin function , 2000, Oncogene.

[38]  L. Lam,et al.  Suppression of Erythroid but Not Megakaryocytic Differentiation of Human K562 Erythroleukemic Cells by Notch-1* , 2000, The Journal of Biological Chemistry.

[39]  J. Sundberg,et al.  Notch signaling is essential for vascular morphogenesis in mice. , 2000, Genes & development.

[40]  Raphael Kopan,et al.  A ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. , 2000, Molecular cell.

[41]  A. Gaultier,et al.  Integrins: Regulators of embryogenesis , 2000, Biology of the cell.

[42]  A. Jarman,et al.  Antagonism of EGFR and notch signalling in the reiterative recruitment of Drosophila adult chordotonal sense organ precursors. , 1999, Development.

[43]  J. Downward,et al.  The small GTP-binding protein R-Ras can influence integrin activation by antagonizing a Ras/Raf-initiated integrin suppression pathway. , 1999, Molecular biology of the cell.

[44]  A. Martinez-Arias,et al.  An activity of Notch regulates JNK signalling and affects dorsal closure in Drosophila , 1999, Current Biology.

[45]  William J. Ray,et al.  A presenilin-1-dependent γ-secretase-like protease mediates release of Notch intracellular domain , 1999, Nature.

[46]  Alan Hall,et al.  Rho GTPases Control Polarity, Protrusion, and Adhesion during Cell Movement , 1999, The Journal of cell biology.

[47]  J. Ramos,et al.  The Death Effector Domain of PEA-15 Is Involved in Its Regulation of Integrin Activation* , 1998, The Journal of Biological Chemistry.

[48]  M. W. Young,et al.  Ligand-induced cleavage and regulation of nuclear entry of Notch in Drosophila melanogaster embryos. , 1998, Genes & development.

[49]  T. Honjo,et al.  Roles of the ankyrin repeats and C-terminal region of the mouse notch1 intracellular region. , 1998, Nucleic acids research.

[50]  U. Lendahl,et al.  Notch and neurogenesis , 1998, Journal of neuroscience research.

[51]  M. Pfaff,et al.  Integrin affinity modulation. , 1998, Trends in cell biology.

[52]  Raphael Kopan,et al.  Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain , 1998, Nature.

[53]  A. Colombatti,et al.  Differential cation regulation of the alpha 5 beta 1 integrin-mediated adhesion of leukemic cells to the central cell-binding domain of fibronectin. , 1997, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[54]  S. Minoguchi,et al.  Involvement of RBP-J in biological functions of mouse Notch1 and its derivatives. , 1997, Development.

[55]  M. Schwartz,et al.  Suppression of Integrin Activation: A Novel Function of a Ras/Raf-Initiated MAP Kinase Pathway , 1997, Cell.

[56]  J. Bos,et al.  Minimal Ras-binding domain of Raf1 can be used as an activation-specific probe for Ras , 1997, Oncogene.

[57]  H. Roehl,et al.  Roles of the RAM and ANK domains in signaling by the C. elegans GLP‐1 receptor. , 1996, The EMBO journal.

[58]  O. Bogler,et al.  Notch signaling inhibits muscle cell differentiation through a CBF1-independent pathway. , 1996, Development.

[59]  J. Boulter,et al.  Expression Patterns ofJagged, Delta1, Notch1, Notch2,andNotch3Genes Identify Ligand–Receptor Pairs That May Function in Neural Development , 1996, Molecular and Cellular Neuroscience.

[60]  Kristiina Vuori,et al.  Integrin Activation by R-ras , 1996, Cell.

[61]  J. Hsieh,et al.  Truncated mammalian Notch1 activates CBF1/RBPJk-repressed genes by a mechanism resembling that of Epstein-Barr virus EBNA2 , 1996, Molecular and cellular biology.

[62]  H. Weintraub,et al.  Signal transduction by activated mNotch: importance of proteolytic processing and its regulation by the extracellular domain. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[63]  Christel Brou,et al.  Signalling downstream of activated mammalian Notch , 1995, Nature.

[64]  J. Rossant,et al.  Notch1 is required for the coordinate segmentation of somites. , 1995, Development.

[65]  M. Fortini,et al.  The suppressor of hairless protein participates in notch receptor signaling , 1994, Cell.

[66]  H. Weintraub,et al.  The intracellular domain of mouse Notch: a constitutively activated repressor of myogenesis directed at the basic helix-loop-helix region of MyoD. , 1994, Development.

[67]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[68]  S. Artavanis-Tsakonas,et al.  Nucleotide sequence from the neurogenic locus Notch implies a gene product that shares homology with proteins containing EGF-like repeats , 1985, Cell.

[69]  Z. Paroush,et al.  EGFR signaling attenuates Groucho-dependent repression to antagonize Notch transcriptional output , 2005, Nature Genetics.

[70]  A. Garcı́a-Bellido,et al.  Activation and function of Notch at the dorsal-ventral boundary of the wing imaginal disc. , 1996, Development.