Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Nov 8;91(23):11089–11093. doi: 10.1073/pnas.91.23.11089

Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator.

F Hofer 1, S Fields 1, C Schneider 1, G S Martin 1
PMCID: PMC45172  PMID: 7972015

Abstract

The yeast two-hybrid system was used to identify proteins that interact with Ras. The H-Ras protein was found to interact with a guanine nucleotide dissociation stimulator (GDS) that has been previously shown to regulate guanine nucleotide exchange on another member of the Ras protein family, Ral. The interaction is mediated by the C-terminal, noncatalytic segment of the RalGDS and can be detected both in vivo, using the two-hybrid system, and in vitro, with purified recombinant proteins. The interaction of the RalGDS C-terminal segment with Ras is specific, dependent on activation of Ras by GTP, and blocked by a mutation that affects Ras effector function. These characteristics are similar to those previously demonstrated for the interaction between Ras and its putative effector, Raf, suggesting that the RalGDS may also be a Ras effector. Consistent with this idea, the RalGDS was found to inhibit the binding of Raf to Ras.

Full text

PDF
11092

Images in this article

11090Image
on p.11090
11091Image
on p.11091
11092Image
on p.11092

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albright C. F., Giddings B. W., Liu J., Vito M., Weinberg R. A. Characterization of a guanine nucleotide dissociation stimulator for a ras-related GTPase. EMBO J. 1993 Jan;12(1):339–347. doi: 10.1002/j.1460-2075.1993.tb05662.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  3. Bielinski D. F., Pyun H. Y., Linko-Stentz K., Macara I. G., Fine R. E. Ral and Rab3a are major GTP-binding proteins of axonal rapid transport and synaptic vesicles and do not redistribute following depolarization stimulated synaptosomal exocytosis. Biochim Biophys Acta. 1993 Sep 19;1151(2):246–256. doi: 10.1016/0005-2736(93)90109-d. [DOI] [PubMed] [Google Scholar]
  4. Boguski M. S., McCormick F. Proteins regulating Ras and its relatives. Nature. 1993 Dec 16;366(6456):643–654. doi: 10.1038/366643a0. [DOI] [PubMed] [Google Scholar]
  5. Bowtell D., Fu P., Simon M., Senior P. Identification of murine homologues of the Drosophila son of sevenless gene: potential activators of ras. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6511–6515. doi: 10.1073/pnas.89.14.6511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chien C. T., Bartel P. L., Sternglanz R., Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. doi: 10.1073/pnas.88.21.9578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duchesne M., Schweighoffer F., Parker F., Clerc F., Frobert Y., Thang M. N., Tocqué B. Identification of the SH3 domain of GAP as an essential sequence for Ras-GAP-mediated signaling. Science. 1993 Jan 22;259(5094):525–528. doi: 10.1126/science.7678707. [DOI] [PubMed] [Google Scholar]
  8. Durfee T., Becherer K., Chen P. L., Yeh S. H., Yang Y., Kilburn A. E., Lee W. H., Elledge S. J. The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. Genes Dev. 1993 Apr;7(4):555–569. doi: 10.1101/gad.7.4.555. [DOI] [PubMed] [Google Scholar]
  9. Field J., Xu H. P., Michaeli T., Ballester R., Sass P., Wigler M., Colicelli J. Mutations of the adenylyl cyclase gene that block RAS function in Saccharomyces cerevisiae. Science. 1990 Jan 26;247(4941):464–467. doi: 10.1126/science.2405488. [DOI] [PubMed] [Google Scholar]
  10. Fields S., Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. doi: 10.1038/340245a0. [DOI] [PubMed] [Google Scholar]
  11. Kataoka T., Powers S., Cameron S., Fasano O., Goldfarb M., Broach J., Wigler M. Functional homology of mammalian and yeast RAS genes. Cell. 1985 Jan;40(1):19–26. doi: 10.1016/0092-8674(85)90304-6. [DOI] [PubMed] [Google Scholar]
  12. Lai C. C., Boguski M., Broek D., Powers S. Influence of guanine nucleotides on complex formation between Ras and CDC25 proteins. Mol Cell Biol. 1993 Mar;13(3):1345–1352. doi: 10.1128/mcb.13.3.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lowy D. R., Willumsen B. M. Function and regulation of ras. Annu Rev Biochem. 1993;62:851–891. doi: 10.1146/annurev.bi.62.070193.004223. [DOI] [PubMed] [Google Scholar]
  14. Martin G. A., Yatani A., Clark R., Conroy L., Polakis P., Brown A. M., McCormick F. GAP domains responsible for ras p21-dependent inhibition of muscarinic atrial K+ channel currents. Science. 1992 Jan 10;255(5041):192–194. doi: 10.1126/science.1553544. [DOI] [PubMed] [Google Scholar]
  15. Moodie S. A., Willumsen B. M., Weber M. J., Wolfman A. Complexes of Ras.GTP with Raf-1 and mitogen-activated protein kinase kinase. Science. 1993 Jun 11;260(5114):1658–1661. doi: 10.1126/science.8503013. [DOI] [PubMed] [Google Scholar]
  16. Munder T., Fürst P. The Saccharomyces cerevisiae CDC25 gene product binds specifically to catalytically inactive ras proteins in vivo. Mol Cell Biol. 1992 May;12(5):2091–2099. doi: 10.1128/mcb.12.5.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Petitjean A., Hilger F., Tatchell K. Comparison of thermosensitive alleles of the CDC25 gene involved in the cAMP metabolism of Saccharomyces cerevisiae. Genetics. 1990 Apr;124(4):797–806. doi: 10.1093/genetics/124.4.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Polakis P., McCormick F. Structural requirements for the interaction of p21ras with GAP, exchange factors, and its biological effector target. J Biol Chem. 1993 May 5;268(13):9157–9160. [PubMed] [Google Scholar]
  19. Powers S., O'Neill K., Wigler M. Dominant yeast and mammalian RAS mutants that interfere with the CDC25-dependent activation of wild-type RAS in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Feb;9(2):390–395. doi: 10.1128/mcb.9.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schiestl R. H., Gietz R. D. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet. 1989 Dec;16(5-6):339–346. doi: 10.1007/BF00340712. [DOI] [PubMed] [Google Scholar]
  21. Shou C., Farnsworth C. L., Neel B. G., Feig L. A. Molecular cloning of cDNAs encoding a guanine-nucleotide-releasing factor for Ras p21. Nature. 1992 Jul 23;358(6384):351–354. doi: 10.1038/358351a0. [DOI] [PubMed] [Google Scholar]
  22. Suzuki N., Tsujino K., Minato T., Nishida Y., Okada T., Kataoka T. Antibody mimicking the action of RAS proteins on yeast adenylyl cyclase: implication for RAS-effector interaction. Mol Cell Biol. 1993 Feb;13(2):769–774. doi: 10.1128/mcb.13.2.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Toda T., Uno I., Ishikawa T., Powers S., Kataoka T., Broek D., Cameron S., Broach J., Matsumoto K., Wigler M. In yeast, RAS proteins are controlling elements of adenylate cyclase. Cell. 1985 Jan;40(1):27–36. doi: 10.1016/0092-8674(85)90305-8. [DOI] [PubMed] [Google Scholar]
  24. Van Aelst L., Barr M., Marcus S., Polverino A., Wigler M. Complex formation between RAS and RAF and other protein kinases. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6213–6217. doi: 10.1073/pnas.90.13.6213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Van Aelst L., Boy-Marcotte E., Camonis J. H., Thevelein J. M., Jacquet M. The C-terminal part of the CDC25 gene product plays a key role in signal transduction in the glucose-induced modulation of cAMP level in Saccharomyces cerevisiae. Eur J Biochem. 1990 Nov 13;193(3):675–680. doi: 10.1111/j.1432-1033.1990.tb19386.x. [DOI] [PubMed] [Google Scholar]
  26. Vojtek A. B., Hollenberg S. M., Cooper J. A. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. doi: 10.1016/0092-8674(93)90307-c. [DOI] [PubMed] [Google Scholar]
  27. Volknandt W., Pevsner J., Elferink L. A., Scheller R. H. Association of three small GTP-binding proteins with cholinergic synaptic vesicles. FEBS Lett. 1993 Feb 8;317(1-2):53–56. doi: 10.1016/0014-5793(93)81490-q. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES