Effector Recognition by the Small GTP-binding Proteins Ras and Ral*

The Ral effector protein RLIP76 (also called RIP/RalBP1) binds to Ral·GTP via a region that shares no sequence homology with the Ras-binding domains of the Ser/Thr kinase c-Raf-1 and the Ral-specific guanine nucleotide exchange factors. Whereas the Ras-binding domains have a similar ubiquitin-like structure, the Ral-binding domain of RLIP was predicted to comprise a coiled-coil region. In order to obtain more information about the specificity and the structural mode of the interaction between Ral and RLIP, we have performed a sequence space and a mutational analysis. The sequence space analysis of a comprehensive nonredundant assembly of Ras-like proteins strongly indicated that positions 36 and 37 in the core of the effector region are tree-determinant positions for all subfamilies of Ras-like proteins and dictate the specificity of the interaction of these GTPases with their effector proteins. Indeed, we could convert the specific interaction with Ras effectors and RLIP by mutating these residues in Ras and Ral. We therefore conclude that positions 36 and 37 are critical for the discrimination between Ras and Ral effectors and that, despite the absence of sequence homology between the Ral-binding and the Ras-binding domains, their mode of interaction is most probably similar.

[1]  Miguel A. Andrade-Navarro,et al.  Automated genome sequence analysis and annotation , 1999, Bioinform..

[2]  R. Wolthuis,et al.  Structure determination of the Ras-binding domain of the Ral-specific guanine nucleotide exchange factor Rlf. , 1998, Biochemistry.

[3]  M. Schwartz,et al.  An activated Rac mutant functions as a dominant negative for membrane ruffling , 1998, Oncogene.

[4]  A. Valencia,et al.  Shaping of Drosophila Alcohol Dehydrogenase Through Evolution: Relationship with Enzyme Functionality , 1998, Journal of Molecular Evolution.

[5]  S. Ryu,et al.  Activation of phospholipase D1 by direct interaction with ADP‐ribosylation factor 1 and RalA , 1998, FEBS letters.

[6]  R. Wolthuis,et al.  Activation of the Small GTPase Ral in Platelets , 1998, Molecular and Cellular Biology.

[7]  A. Wittinghofer,et al.  Kinetic analysis by fluorescence of the interaction between Ras and the catalytic domain of the guanine nucleotide exchange factor Cdc25Mm. , 1998, Biochemistry.

[8]  S. Beck,et al.  TAPASIN, DAXX, RGL2, HKE2 and four new genes (BING 1, 3 to 5) form a dense cluster at the centromeric end of the MHC. , 1998, Journal of molecular biology.

[9]  R. Wolthuis,et al.  Ras-dependent activation of the small GTPase Ral , 1998, Current Biology.

[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]  S. Yokoyama,et al.  Interactions of the Amino Acid Residue at Position 31 of the c-Ha-Ras Protein with Raf-1 and RalGDS* , 1998, The Journal of Biological Chemistry.

[12]  K. Jakobs,et al.  Specific Inhibition of Phorbol Ester-stimulated Phospholipase D by Clostridium sordellii Lethal Toxin and Clostridium difficile Toxin B-1470 in HEK-293 Cells , 1998, The Journal of Biological Chemistry.

[13]  T. Urano,et al.  An Eps Homology (EH) Domain Protein That Binds to the Ral-GTPase Target, RalBP1* , 1997, The Journal of Biological Chemistry.

[14]  R. Wolthuis,et al.  Stimulation of gene induction and cell growth by the Ras effector Rlf , 1997, The EMBO journal.

[15]  Hans Robert Kalbitzer,et al.  Structure of the Ras-binding domain of RalGEF and implications for Ras binding and signalling , 1997, Nature Structural Biology.

[16]  Lan Huang,et al.  Three-dimensional structure of the Ras-interacting domain of RalGDS , 1997, Nature Structural Biology.

[17]  A. Kikuchi,et al.  The post‐translational modifications of Ral and Rac1 are important for the action of Ral‐binding protein 1, a putative effector protein of Ral , 1997, FEBS letters.

[18]  S. Hammond,et al.  RalA interacts directly with the Arf-responsive, PIP2-dependent phospholipase D1. , 1997, Biochemical and biophysical research communications.

[19]  B. Roufogalis,et al.  Identification and Characterization of a Calmodulin-binding Domain in Ral-A, a Ras-related GTP-binding Protein Purified from Human Erythrocyte Membrane* , 1997, The Journal of Biological Chemistry.

[20]  Y. Matsuura,et al.  Characterization of Ral GDP Dissociation Stimulator-like (RGL) Activities to Regulate c-fos Promoter and the GDP/GTP Exchange of Ral* , 1997, The Journal of Biological Chemistry.

[21]  A. Pellicer,et al.  rsc: a novel oncogene with structural and functional homology with the gene family of exchange factors for Ral , 1997, Oncogene.

[22]  M. White,et al.  RalGDS Functions in Ras- and cAMP-mediated Growth Stimulation* , 1997, The Journal of Biological Chemistry.

[23]  T. Kataoka,et al.  Synergistic activation of c-fos promoter activity by Raf and Ral GDP dissociation stimulator , 1997, Oncogene.

[24]  S. Peterson,et al.  Identification of a Novel RalGDS-related Protein as a Candidate Effector for Ras and Rap1* , 1996, Journal of Biological Chemistry.

[25]  Ralf Janknecht,et al.  Ras/Rap effector specificity determined by charge reversal , 1996, Nature Structural Biology.

[26]  A. Wittinghofer,et al.  Interactions between Ras proteins and their effectors. , 1996, Current opinion in biotechnology.

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

[28]  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.

[29]  R. Bhullar,et al.  Characterization of human platelet GTPase activating protein for the Ral GTP-binding protein. , 1996, Biochimica et biophysica acta.

[30]  J. Downward,et al.  Activation of phosphoinositide 3‐kinase by interaction with Ras and by point mutation. , 1996, The EMBO journal.

[31]  Gudrun Horn,et al.  Differential Interaction of the Ras Family GTP-binding Proteins H-Ras, Rap1A, and R-Ras with the Putative Effector Molecules Raf Kinase and Ral-Guanine Nucleotide Exchange Factor , 1996, The Journal of Biological Chemistry.

[32]  Anthony J. Muslin,et al.  Ras‐interacting domain of RGL blocks Ras‐dependent signal transduction in Xenopus oocytes , 1996, FEBS letters.

[33]  T. Urano,et al.  Ral‐GTPases mediate a distinct downstream signaling pathway from Ras that facilitates cellular transformation. , 1996, The EMBO journal.

[34]  J. Camonis,et al.  p 95 vav Associates with the Nuclear Protein Ku-70 , 1995 .

[35]  Park Sh,et al.  A putative effector of Ral has homology to Rho/Rac GTPase activating proteins. , 1995 .

[36]  Zhimin Lu,et al.  Involvement of Ral GTPase in v-Src-induced phospholipase D activation , 1995, Nature.

[37]  J. Camonis,et al.  Bridging Ral GTPase to Rho Pathways , 1995, The Journal of Biological Chemistry.

[38]  A. Wittinghofer,et al.  The 2.2 Å crystal structure of the Ras-binding domain of the serine/threonine kinase c-Raf1 in complex with RaplA and a GTP analogue , 1995, Nature.

[39]  D. Fry,et al.  Solution structure of the Ras-binding domain of c-Raf-1 and identification of its Ras interaction surface. , 1995, Biochemistry.

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

[41]  Alfred Wittinghofer,et al.  Quantitative Analysis of the Complex between p21 and the Ras-binding Domain of the Human Raf-1 Protein Kinase (*) , 1995, The Journal of Biological Chemistry.

[42]  C. Sander,et al.  A method to predict functional residues in proteins , 1995, Nature Structural Biology.

[43]  J. Bischoff,et al.  Identification of the guanine nucleotide dissociation stimulator for Ral as a putative effector molecule of R-ras, H-ras, K-ras, and Rap. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[44]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[45]  S. Fields,et al.  Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[46]  S. Demo,et al.  ralGDS family members interact with the effector loop of ras p21 , 1994, Molecular and cellular biology.

[47]  M. Shirouzu,et al.  Mutations that abolish the ability of Ha-Ras to associate with Raf-1. , 1994, Oncogene.

[48]  Véronique Picard,et al.  A rapid and efficient one-tube PCR-based mutagenesis technique using Pfu DNA polymerase , 1994, Nucleic Acids Res..

[49]  T. Saito,et al.  Purification of glutathione S-transferase fusion proteins as a non-degraded form by using a protease-negative E. coli strain, AD202. , 1994, Nucleic acids research.

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

[51]  I. Macara,et al.  Ral and Rab3a are major GTP-binding proteins of axonal rapid transport and synaptic vesicles and do not redistribute following depolarization stimulated synaptosomal exocytosis. , 1993, Biochimica et biophysica acta.

[52]  G. Wildey,et al.  Isolation of cDNA clones and tissue expression of rat ral A and ral B GTP-binding proteins. , 1993, Biochemical and biophysical research communications.

[53]  R. Scheller,et al.  Association of three small GTP‐binding proteins with cholinergic synaptic vesicles , 1993, FEBS letters.

[54]  R. Weinberg,et al.  Characterization of a guanine nucleotide dissociation stimulator for a ras‐related GTPase. , 1993, The EMBO journal.

[55]  S. Henikoff,et al.  Amino acid substitution matrices from protein blocks. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[56]  R. Bhullar.  Identification of some of the brain Gn27 as the ral gene product Comparison between the brain and platelet Gn‐proteins , 1992, FEBS letters.

[57]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[58]  L. Feig,et al.  Characterization of a GTPase-activating protein for the Ras-related Ral protein. , 1991, The Journal of biological chemistry.

[59]  I. Schlichting,et al.  Guanine nucleotide binding properties of the mammalian RalA protein produced in Escherichia coli. , 1990, The Journal of biological chemistry.

[60]  R. Haslam,et al.  Identification of multiple ral gene products in human platelets that account for some but not all of the platelet Gn‐proteins , 1990, FEBS letters.

[61]  R. Snyderman,et al.  Identification of the ral and rac1 gene products, low molecular mass GTP-binding proteins from human platelets. , 1989, The Journal of biological chemistry.

[62]  P. Chardin,et al.  Coding sequences of human ralA and ralB cDNAs. , 1989, Nucleic acids research.

[63]  B. Olofsson,et al.  Expression of the ras-related ralA, rho12 and rab genes in adult mouse tissues. , 1988, Oncogene.

[64]  P. Chardin,et al.  The ral gene: a new ras related gene isolated by the use of a synthetic probe. , 1986, The EMBO journal.

[65]  W. Kabsch,et al.  Dictionary of protein secondary structure: Pattern recognition of hydrogen‐bonded and geometrical features , 1983, Biopolymers.

[66]  Gerald R. Fink,et al.  Methods in Yeast Genetics: A Laboratory Course Manual , 1987 .

[67]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[68]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[69]  S. Cantor,et al.  Identification and Characterization of Ral-Binding Protein 1, a Potential Downstream Target of Ral GTPases , 2022 .