Differential Structural Requirements for Interaction of Ras Protein with Its Distinct Downstream Effectors
Open accessAbstract
Ras proteins have multiple effectors of distinct structures that do not share significant structural homology at their Ras interaction sites. To prove possible differences in their recognition mechanisms of Ras, we screened 44 human Ha-Ras proteins carrying mutations in the effector region and its flanking sequences for interaction with human Raf-1, Schizosaccharomyces pombe Byr2, and Saccharomyces cerevisiae adenylyl cyclase. The Ras binding specificities were largely shared between Raf-1 and Byr2 although Ras mutants, Y32F, T35S, and A59E, had their affinities for Byr2 selectively reduced. The only exception was Ras(D38N), which lost the ability to bind Raf-1 while retaining the activity to bind Byr2 and complement the Byr2- phenotype of S. pombe. On the other hand, adenylyl cyclase had quite distinct requirements for Ras residues; mutations P34G and T58A selectively abolished the ability to bind and activate it without considerably affecting the interaction with Raf-1 and Byr2. Y32F mutant, whereas losing the ability to activate Raf-1 and Byr2, could activate adenylyl cyclase efficiently. In addition, V45E mutation was found to impair the ability of Ras to activate both Raf-1 and adenylyl cyclase without significantly affecting the binding affinities for them. These results demonstrate that significant differences exist in the recognition mechanisms by which the three effector molecules associate with Ras and suggest that a region of Ras required for activation of the effectors in general may exist separately from that for binding the effectors.
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Activated RAS proteins can interact with many downstream effector proteins, such as RAF kinase, son of sevenless (SOS), phosphatidylinositol 3-kinase (PI3K) and Ral guanine nucleotide dissociation stimulating factors (RalGDS), which regulate cell events such as cell proliferation and apoptosis.
“Activated RAS proteins can interact with many downstream effector proteins, such as RAF kinase, son of sevenless (SOS), phosphatidylinositol 3-kinase (PI3K) and Ral guanine nucleotide dissociation stimulating factors (RalGDS), which regulate cell events such as cell proliferation and apoptosis.323–325”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [323]
Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961 · see citing works
Saiki, R. K., Scharf, S., Faloona, F., Mullis, K. B., Horn, G. T., Erlich, H. A., & Arnheim, N. (1985) · Science · doi:10.1126/science.2999980 · Cited by 8,956 · see citing works
Transformation of intact yeast cells treated with alkali cations
Ito, H., Fukuda, Y., Murata, K., & Kimura, A. (1983) · Journal of Bacteriology · doi:10.1128/jb.153.1.163-168.1983 · Cited by 7,518 · see citing works
Smith, D. B., & Johnson, K. S. (1988) · Gene · doi:10.1016/0378-1119(88)90005-4 · Cited by 5,974 · see citing works
Mammalian Ras interacts directly with the serine/threonine kinase Raf.
Vojtek, A., Hollenberg, S., & Cooper, J. A. (1993) · Cell · doi:10.1016/0092-8674(93)90307-c · Cited by 1,971 · see citing works
Phosphatidylinositol-3-OH kinase direct target of Ras
Rodriguez-Viciana, P., Warne, P. H., Dhand, R., Vanhaesebroeck, B., Gout, I., Fry, M. J., Waterfield, M. D., & Downward, J. (1994) · Nature · doi:10.1038/370527a0 · Cited by 1,954 · see citing works
PAI, E. F., Krengel, U., Petsko, G. A., Goody, R. S., Kabsch, W., & Wittinghofer, A. (1990) · The EMBO Journal · doi:10.1002/j.1460-2075.1990.tb07409.x · Cited by 1,074 · see citing works
Milburn, M. V., Tong, L., deVos, A. M., Brünger, A. T., Yamaizumi, Z., Nishimura, S., & Kim, S. (1990) · Science · doi:10.1126/science.2406906 · Cited by 1,062 · see citing works
Complexes of Ras⋅GTP with Raf-1 and Mitogen-Activated Protein Kinase Kinase
Moodie, S. A., Willumsen, B. M., Weber, M. J., & Wolfman, A. (1993) · Science · doi:10.1126/science.8503013 · Cited by 968 · see citing works
A ras-related gene with transformation suppressor activity
Kitayama, H., Sugimoto, Y., Matsuzaki, T., Ikawa, Y., & Noda, M. (1989) · Cell · doi:10.1016/0092-8674(89)90985-9 · Cited by 942 · see citing works
Normal and oncogenic p21ras proteins bind to the amino-terminal regulatory domain of c-Raf-1
Zhang, X., Settleman, J. E., Kyriakis, J. M., Takeuchi-Suzuki, E., Elledge, S. J., Marshall, M. S., Bruder, J. T., Rapp, U. R., & Avruch, J. (1993) · Nature · doi:10.1038/364308a0 · Cited by 859 · see citing works
nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine.
Maundrell, K. (1990) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)38525-4 · Cited by 841 · see citing works
Direct interaction of Ras and the amino-terminal region of Raf-1 in vitro
Warne, P. H., Vician, P. R., & Downward, J. (1993) · Nature · doi:10.1038/364352a0 · Cited by 729 · see citing works
Multiple ras functions can contribute to mammalian cell transformation
White, M. A., Nicolette, C. A., Minden, A., Polverino, A. J., Aelst, L. V., Karin, M., & Wigler, M. H. (1995) · Cell · doi:10.1016/0092-8674(95)90507-3 · Cited by 650 · see citing works
Nassar, N. N., Horn, G., Herrmann, C., Scherer, A., McCormick, F. P., & Wittinghofer, A. (1995) · Nature · doi:10.1038/375554a0 · Cited by 631 · see citing works
Complex formation between RAS and RAF and other protein kinases.
Aelst, L. V., Barr, M. M., Marcus, S., Polverino, A. J., & Wigler, M. H. (1993) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.90.13.6213 · Cited by 623 · see citing works
Raf meets Ras: completing the framework of a signal transduction pathway
Avruch, J., Zhang, X.-F., & Kyriakis, J. M. (1994) · Trends in Biochemical Sciences · doi:10.1016/0968-0004(94)90005-1 · Cited by 577 · see citing works
The ins and outs of Raf kinases
Daum, G., Eisenmann-Tappe, I., Fries, H.-W., Troppmair, J., & Rapp, U. R. (1994) · Trends in Biochemical Sciences · doi:10.1016/0968-0004(94)90133-3 · Cited by 524 · see citing works
Okazaki, K., Okazaki, N., Kume, K., Jinno, S., Tanaka, K., & Okayama, H. (1990) · Nucleic Acids Research · doi:10.1093/nar/18.22.6485 · Cited by 426 · see citing works
DNA sequence and characterization of the S. cerevisiae gene encoding adenylate cyclase
Kataoka, T., Broek, D., & Wigler, M. H. (1985) · Cell · doi:10.1016/0092-8674(85)90179-5 · Cited by 425 · see citing works
Self-citationKrengel, U., Schlichting, I., Scherer, A., Schumann, R., Frech, M., John, J., Kabsch, W., PAI, E. F., & Wittinghofer, A. (1990) · Cell · doi:10.1016/0092-8674(90)90018-a · Cited by 386 · see citing works
Cellular interactions in development : a practical approach
Hartley, D. A. (1993) · Oxford University Press eBooks · Cited by 378 · see citing works
Pizon, V., Chardin, P., Janoueix‐Lerosey, I., Olofsson, B., & Tavitian, A. (1988) · PubMed · Cited by 336 · see citing works
Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator.
Hofer, F., Fields, S., Schneider, C., & Martin, G. S. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.23.11089 · Cited by 287 · see citing works
Identification of effector residues and a neutralizing epitope of Ha-ras-encoded p21.
Sigal, I. S., Gibbs, W. W., D'Alonzo, J., & Scolnick, E. M. (1986) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.83.13.4725 · Cited by 285 · see citing works
Spaargaren, M., & Bischoff, J. R. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.26.12609 · Cited by 265 · see citing works
Role of a ras homolog in the life cycle of schizosaccharomyces pombe
Fukui, Y., Kozasa, T., Kaziro, Y., Takeda, T., & Yamamoto, M. (1986) · Cell · doi:10.1016/0092-8674(86)90767-1 · Cited by 265 · see citing works
Self-citationPremeiotic DNA synthesis in fission yeast
Egel, R., & EGEL-MITANI, M. (1974) · Experimental Cell Research · doi:10.1016/0014-4827(74)90626-0 · Cited by 263 · see citing works
ralGDS family members interact with the effector loop of ras p21.
Kikuchi, A., Demo, S. D., Ye, Z. H., Chen, Y. W., & Williams, L. T. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.11.7483 · Cited by 259 · see citing works
Self-citationA conserved kinase cascade for MAP kinase activation in yeast
Errede, B., & Levin, D. E. (1993) · Current Opinion in Cell Biology · doi:10.1016/0955-0674(93)90112-4 · Cited by 251 · see citing works
Two Distinct Raf Domains Mediate Interaction with Ras
Brtva, T. R., Drugan, J. K., Ghosh, S., Terrell, R. S., Campbell, S. L., Bell, R. M., & Der, C. J. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.17.9809 · Cited by 247 · see citing works
Evidence for the in vitro and in vivo interaction of Ras with protein kinase C zeta.
Díaz-Meco, M. T., Lozano, J., Municio, M. M., Berra, E., Frutos, S., Sanz, L., & Moscat, J. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)31753-8 · Cited by 220 · see citing works
Wang, Y., Xu, H. P., Riggs, M. G., et al. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.7.3554 · Cited by 209 · see citing works
The ras oncogene--an important regulatory element in lower eucaryotic organisms
Gibbs, W. W., & Marshall, M. S. (1989) · Microbiological Reviews · doi:10.1128/mr.53.2.171-185.1989 · Cited by 205 · see citing works
Mutational analysis of a ras catalytic domain.
Willumsen, B. M., Papageorge, A. G., Kung, H. F., Bekesi, E., Robins, T., Johnsen, M., Vass, W. C., & Lowy, D. R. (1986) · Molecular and Cellular Biology · doi:10.1128/mcb.6.7.2646 · Cited by 195 · see citing works
The effector interactions of p21ras
Marshall, M. S. (1993) · Trends in Biochemical Sciences · doi:10.1016/0968-0004(93)90175-m · Cited by 191 · see citing works
Chuang, E., Barnard, D., Hettich, L., Zhang, X. F., Avruch, J., & Marshall, M. S. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.8.5318 · Cited by 184 · see citing works
Koide, H., Satoh, T., Nakafuku, M., & Kaziro, Y. (1993) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.90.18.8683 · Cited by 176 · see citing works
Hu, C., Kariya, K., Tamada, M., Akasaka, K., Shirouzu, M., Yokoyama, S., & Kataoka, T. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.51.30274 · Cited by 153 · see citing works
Self-citationThe Effect of Posttranslational Modifications on the Interaction of Ras2 with Adenylyl Cyclase
Kuroda, Y., Suzuki, N., & Kataoka, T. (1993) · Science · doi:10.1126/science.8430318 · Cited by 143 · see citing works
Self-citationInvolvement of ras in sexual differentiation but not in growth control in fission yeast
Nadin‐Davis, S. A., Nasim, A., & Beach, D. (1986) · The EMBO Journal · doi:10.1002/j.1460-2075.1986.tb04593.x · Cited by 138 · see citing works
Suzuki, N., Choe, H. R., Nishida, Y., Yamawaki‐Kataoka, Y., Ohnishi, S., Tamaoki, T., & Kataoka, T. (1990) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.87.22.8711 · Cited by 128 · see citing works
Self-citationProtein Kinase Byr2 Is a Target of Ras1 in the Fission Yeast Schizosaccharomyces pombe
Masuda, T., Kariya, K., Shinkai, M., Okada, T., & Kataoka, T. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.5.1979 · Cited by 73 · see citing works
Self-citationAssociation of MEK1 with p21 ras GMPPNP Is Dependent on B-Raf
Moodie, S. A., Paris, M. J., Kölch, W., & Wolfman, A. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.11.7153 · Cited by 73 · see citing works
The post-translational modification of ras p21 is important for Raf-1 activation.
Kikuchi, A., & Williams, L. T. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)32126-9 · Cited by 69 · see citing works
Self-citationMutations that abolish the ability of Ha-Ras to associate with Raf-1.
Shirouzu, M., Koide, H., Fujita‐Yoshigaki, J., Oshio, H., Toyama, Y., Yamasaki, K., Fuhrman, S. A., Villafranca, E., Kaziro, Y., & Yokoyama, S. (1994) · PubMed · Cited by 57 · see citing works
Self-citationGhosh, S., & Bell, R. M. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)47350-4 · Cited by 56 · see citing works
Identification of amino acid residues required for Ras p21 target activation.
Marshall, M. S., Davis, L. J., Keys, R. D., et al. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.8.3997 · Cited by 54 · see citing works
Nur-e-Kamal, M. S. A., Sizeland, A. M., D’Abaco, G. M., & Maruta, H. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)45958-3 · Cited by 53 · see citing works
Raf-1 N-Terminal Sequences Necessary for Ras-Raf Interaction and Signal Transduction
Pumiglia, K. M., Chow, Y., Fabian, J. R., et al. (1995) · Molecular and Cellular Biology · doi:10.1128/mcb.15.1.398 · Cited by 50 · see citing works
Minato, T., Wang, J., Akasaka, K., Okada, T., Suzuki, N., & Kataoka, T. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)31899-9 · Cited by 45 · see citing works
Self-citationp21-ras effector domain mutants constructed by "cassette" mutagenesis.
Stone, J. C., Vass, W. C., Willumsen, B. M., & Lowy, D. R. (1988) · Molecular and Cellular Biology · doi:10.1128/mcb.8.8.3565 · Cited by 43 · see citing works
Yamasaki, K., Kawai, G., Ito, Y., Muto, Y., Fujita‐Yoshigaki, J., Miyazawa, T., Nishimura, S., & Yokoyama, S. (1989) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(89)90780-8 · Cited by 30 · see citing works
Self-citationShirouzu, M., Fujita‐Yoshigaki, J., Ito, Y., et al. (1992) · PubMed · Cited by 29 · see citing works
Self-citationYamasaki, K., Shirouzu, M., Muto, Y., Fujita‐Yoshigaki, J., Koide, H., Ito, Y., Kawai, G., Hattori, S., & Yokoyama, S. (1994) · Biochemistry · doi:10.1021/bi00167a009 · Cited by 26 · see citing works
Self-citationA Constitutive Effector Region on the C-terminal Side of Switch I of the Ras Protein
Fujita‐Yoshigaki, J., Shirouzu, M., Ito, Y., Hattori, S., Furuyama, S., Nishimura, S., & Yokoyama, S. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.9.4661 · Cited by 24 · see citing works
Self-citationFujita‐Yoshigaki, J., Shirouzu, M., Koide, H., Nishimura, S., & Yokoyama, S. (1991) · FEBS Letters · doi:10.1016/0014-5793(91)80665-p · Cited by 13 · see citing works
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- Authors
- Akasaka, K., Tamada, M., Wang, F., Kariya, K.-I., Shima, F., Kikuchi, A., Yamamoto, M., Shirouzu, M., Yokoyama, S., & Kataoka, T.
- Affiliations
- Kobe University (JP)
- Hiroshima University (JP)
- The University of Tokyo (JP)
- RIKEN (JP)
- Corresponding author
- Kataoka, Tohru
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1996-03
- Volume
- 271
- Issue
- 10
- Pages
- 5353-5360
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2083260280
- Cited by
- 69
- References
- 57
- Citations by year
- 2025: 2 · 2023: 2 · 2019: 2 · 2018: 1 · 2016: 2 · 2014: 1 · 2013: 1 · 2012: 1
- FWCI
- 3.41
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- Ras proteins, Ras effectors, Raf-1, adenylate cyclase, RAS mutations
- MeSH terms
- Adenylyl Cyclases, Amino Acid Sequence, Antibodies, Monoclonal, Base Sequence, Binding Sites, Genetic Complementation Test, Humans, Kinetics, Molecular Sequence Data, Proto-Oncogene Proteins, Saccharomyces cerevisiae, Schizosaccharomyces, Structure-Activity Relationship, Polymerase Chain Reaction, Proto-Oncogene Proteins p21(ras), Mutagenesis, Site-Directed, Protein Serine-Threonine Kinases, Point Mutation, DNA Primers, ras Proteins, Proto-Oncogene Proteins c-raf
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