M-Ras/R-Ras3, a Transforming Ras Protein Regulated by Sos1, GRF1, and p120 Ras GTPase-activating Protein, Interacts with the Putative Ras Effector AF6*

M-Ras is a Ras-related protein that shares ∼55% identity with K-Ras and TC21. The M-Ras message was widely expressed but was most predominant in ovary and brain. Similarly to Ha-Ras, expression of mutationally activated M-Ras in NIH 3T3 mouse fibroblasts or C2 myoblasts resulted in cellular transformation or inhibition of differentiation, respectively. M-Ras only weakly activated extracellular signal-regulated kinase 2 (ERK2), but it cooperated with Raf, Rac, and Rho to induce transforming foci in NIH 3T3 cells, suggesting that M-Ras signaled via alternate pathways to these effectors. Although the mitogen-activated protein kinase/ERK kinase inhibitor, PD98059, blocked M-Ras-induced transformation, M-Ras was more effective than an activated mitogen-activated protein kinase/ERK kinase mutant at inducing focus formation. These data indicate that multiple pathways must contribute to M-Ras-induced transformation. M-Ras interacted poorly in a yeast two-hybrid assay with multiple Ras effectors, including c-Raf-1, A-Raf, B-Raf, phosphoinositol-3 kinase δ, RalGDS, and Rin1. Although M-Ras coimmunoprecipitated with AF6, a putative regulator of cell junction formation, overexpression of AF6 did not contribute to fibroblast transformation, suggesting the possibility of novel effector proteins. The M-Ras GTP/GDP cycle was sensitive to the Ras GEFs, Sos1, and GRF1 and to p120 Ras GAP. Together, these findings suggest that while M-Ras is regulated by similar upstream stimuli to Ha-Ras, novel targets may be responsible for its effects on cellular transformation and differentiation.

[1]  Channing J Der,et al.  Increasing complexity of Ras signaling , 1998, Oncogene.

[2]  J. Hancock,et al.  Ras Isoforms Vary in Their Ability to Activate Raf-1 and Phosphoinositide 3-Kinase* , 1998, The Journal of Biological Chemistry.

[3]  T. Pawson,et al.  PDZ-domain-mediated interaction of the Eph-related receptor tyrosine kinase EphB3 and the ras-binding protein AF6 depends on the kinase activity of the receptor. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Malumbres,et al.  RAS pathways to cell cycle control and cell transformation. , 1998, Frontiers in bioscience : a journal and virtual library.

[5]  John Kuriyan,et al.  The structural basis of the activation of Ras by Sos , 1998, Nature.

[6]  F. Hobbs,et al.  Identification of a Novel Inhibitor of Mitogen-activated Protein Kinase Kinase* , 1998, The Journal of Biological Chemistry.

[7]  M. White,et al.  A Role for RalGDS and a Novel Ras Effector in the Ras-mediated Inhibition of Skeletal Myogenesis* , 1998, The Journal of Biological Chemistry.

[8]  F. Ribeiro-Neto,et al.  Mitogenic and oncogenic properties of the small G protein Rap1b. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[9]  B. Wasylyk,et al.  Ets transcription factors: nuclear effectors of the Ras-MAP-kinase signaling pathway. , 1998, Trends in biochemical sciences.

[10]  R. Kahn,et al.  Functional association between Arf and RalA in active phospholipase D complex. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Avruch,et al.  Identification of Nore1 as a Potential Ras Effector* , 1998, The Journal of Biological Chemistry.

[12]  A. Wolfman,et al.  Ha-ras and N-ras regulate MAPK activity by distinct mechanisms in vivo , 1998, Oncogene.

[13]  A. Hall,et al.  Rho GTPases and the actin cytoskeleton. , 1998, Science.

[14]  J. Jackson,et al.  Ras-GRF Activates Ha-Ras, but Not N-Ras or K-Ras 4B, Proteinin Vivo * , 1998, The Journal of Biological Chemistry.

[15]  E. Friedman,et al.  Oncogenic Ki-ras but Not Oncogenic Ha-rasBlocks Integrin β1-Chain Maturation in Colon Epithelial Cells* , 1997, The Journal of Biological Chemistry.

[16]  A. Kimmelman,et al.  Identification and characterization of R-ras3: a novel member of the RAS gene family with a non-ubiquitous pattern of tissue distribution , 1997, Oncogene.

[17]  S. Grant,et al.  A role for the Ras signalling pathway in synaptic transmission and long-term memory , 1997, Nature.

[18]  Ken Matsumoto,et al.  Novel small GTPase M-Ras participates in reorganization of actin cytoskeleton , 1997, Oncogene.

[19]  K. Kaibuchi,et al.  The Ras Target AF-6 Interacts with ZO-1 and Serves as a Peripheral Component of Tight Junctions in Epithelial Cells , 1997, The Journal of cell biology.

[20]  M. Itoh,et al.  Afadin: A Novel Actin Filament–binding Protein with One PDZ Domain Localized at Cadherin-based Cell-to-Cell Adherens Junction , 1997, The Journal of cell biology.

[21]  R. Kucherlapati,et al.  K-ras is an essential gene in the mouse with partial functional overlap with N-ras. , 1997, Genes & development.

[22]  Jun Miyoshi,et al.  K-Ras is essential for the development of the mouse embryo , 1997, Oncogene.

[23]  M. Matsuda,et al.  Activation of R-Ras by Ras-Guanine Nucleotide-releasing Factor* , 1997, The Journal of Biological Chemistry.

[24]  M. White,et al.  Signaling through mitogen-activated protein kinase and Rac/Rho does not duplicate the effects of activated Ras on skeletal myogenesis , 1997, Molecular and cellular biology.

[25]  F. McCormick,et al.  Cdc42 regulates anchorage-independent growth and is necessary for Ras transformation , 1997, Molecular and cellular biology.

[26]  M. White,et al.  Protein binding and signaling properties of RIN1 suggest a unique effector function. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[27]  P. Warne,et al.  Role of Phosphoinositide 3-OH Kinase in Cell Transformation and Control of the Actin Cytoskeleton by Ras , 1997, Cell.

[28]  M. Su,et al.  Role of p38 and JNK mitogen-activated protein kinases in the activation of ternary complex factors , 1997, Molecular and cellular biology.

[29]  C. Der,et al.  The Ras-related Protein Rheb Is Farnesylated and Antagonizes Ras Signaling and Transformation* , 1997, The Journal of Biological Chemistry.

[30]  H. Yao,et al.  cAMP Activates MAP Kinase and Elk-1 through a B-Raf- and Rap1-Dependent Pathway , 1997, Cell.

[31]  P. Warne,et al.  R-Ras can activate the phosphoinositide 3-kinase but not the MAP kinase arm of the Ras effector pathways , 1997, Current Biology.

[32]  A. Wittinghofer,et al.  How Ras-related proteins talk to their effectors. , 1996, Trends in biochemical sciences.

[33]  Jonathan A. Cooper,et al.  TC21 causes transformation by Raf-independent signaling pathways , 1996, Molecular and cellular biology.

[34]  M. Wigler,et al.  A Role for the Ral Guanine Nucleotide Dissociation Stimulator in Mediating Ras-induced Transformation* , 1996, The Journal of Biological Chemistry.

[35]  M. Wigler,et al.  Oncogenic Ras activation of Raf/mitogen-activated protein kinase-independent pathways is sufficient to cause tumorigenic transformation , 1996, Molecular and cellular biology.

[36]  C. Der,et al.  Involvement of the Switch 2 Domain of Ras in Its Interaction with Guanine Nucleotide Exchange Factors (*) , 1996, The Journal of Biological Chemistry.

[37]  D. Bar-Sagi,et al.  The Ras superfamily of GTPases 1 , 1996, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

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

[39]  M. Wigler,et al.  Stimulation of Membrane Ruffling and MAP Kinase Activation by Distinct Effectors of RAS , 1996, Science.

[40]  M. Nakafuku,et al.  Identification of AF-6 and Canoe as Putative Targets for Ras (*) , 1996, The Journal of Biological Chemistry.

[41]  C. Der,et al.  Overexpression of the Ras-related TC21/R-Ras2 protein may contribute to the development of human breast cancers. , 1996, Oncogene.

[42]  C. Der,et al.  Activation of Rac1, RhoA, and mitogen-activated protein kinases is required for Ras transformation , 1995, Molecular and cellular biology.

[43]  S. Aaronson,et al.  A novel insertional mutation in the TC21 gene activates its transforming activity in a human leiomyosarcoma cell line. , 1995, Oncogene.

[44]  C. Der,et al.  Tyrosine phosphorylation regulates the adhesions of ras-transformed breast epithelia , 1995, The Journal of cell biology.

[45]  John Calvin Reed,et al.  R-Ras promotes apoptosis caused by growth factor deprivation via a Bcl- 2 suppressible mechanism , 1995, The Journal of cell biology.

[46]  C. Der,et al.  Guanine nucleotide exchange factors: Activators of the Ras superfamily of proteins , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.

[47]  F. McCormick,et al.  An essential role for Rac in Ras transformation , 1995, Nature.

[48]  M. Wigler,et al.  Multiple ras functions can contribute to mammalian cell transformation , 1995, Cell.

[49]  A. Vojtek,et al.  Ras-Raf interaction: two-hybrid analysis. , 1995, Methods in enzymology.

[50]  C. Der,et al.  Biological assays for Ras transformation. , 1995, Methods in enzymology.

[51]  A. Ashworth,et al.  Assay and expression of mitogen-activated protein kinase, MAP kinase kinase, and Raf. , 1995, Methods in enzymology.

[52]  C. Hauser,et al.  Ras-mediated transcription activation: analysis by transient cotransfection assays. , 1995, Methods in enzymology.

[53]  C. Der,et al.  R-Ras induces malignant, but not morphologic, transformation of NIH3T3 cells. , 1994, Oncogene.

[54]  Michael Karin,et al.  Membrane targeting of the nucleotide exchange factor Sos is sufficient for activating the Ras signaling pathway , 1994, Cell.

[55]  C. Der,et al.  Membrane-targeting potentiates guanine nucleotide exchange factor CDC25 and SOS1 activation of Ras transforming activity. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[56]  N. Ahn,et al.  Transformation of mammalian cells by constitutively active MAP kinase kinase. , 1994, Science.

[57]  S. Aaronson,et al.  A human oncogene of the RAS superfamily unmasked by expression cDNA cloning. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[58]  C. Der,et al.  Activated or dominant inhibitory mutants of Rap1A decrease the oxidative burst of Epstein-Barr virus-transformed human B lymphocytes. , 1994, The Journal of biological chemistry.

[59]  C. Marshall,et al.  Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells , 1994, Cell.

[60]  Mark S. Boguski,et al.  Proteins regulating Ras and its relatives , 1993, Nature.

[61]  P. Nowell,et al.  Cloning of the ALL-1 fusion partner, the AF-6 gene, involved in acute myeloid leukemias with the t(6;11) chromosome translocation. , 1993, Cancer research.

[62]  J. Bischoff,et al.  Bcl-2 associates with the ras-related protein R-rasp23 , 1993, Nature.

[63]  Julian Downward,et al.  Epidermal growth factor regulates p21 ras through the formation of a complex of receptor, Grb2 adapter protein, and Sos nucleotide exchange factor , 1993, Cell.

[64]  R. Treisman,et al.  The SRF accessory protein Elk-1 contains a growth factor-regulated transcriptional activation domain , 1993, Cell.

[65]  A. Jesaitis,et al.  Inhibition of Rap1A binding to cytochrome b558 of NADPH oxidase by phosphorylation of Rap1A. , 1991, Science.

[66]  D. Lowy,et al.  Identification of small clusters of divergent amino acids that mediate the opposing effects of ras and Krev-1. , 1990, Science.

[67]  W. Herr,et al.  Differential transcriptional activation by Oct-1 and Oct-2: Interdependent activation domains induce Oct-2 phosphorylation , 1990, Cell.

[68]  A. Pellicer,et al.  Differential expression of the ras gene family in mice , 1987, Molecular and cellular biology.

[69]  C. Der,et al.  Biological and biochemical properties of human ras H genes mutated at codon 61 , 1986, Cell.

[70]  C. Cepko,et al.  Construction and applications of a highly transmissible murine retrovirus shuttle vector , 1984, Cell.