1994
DOI: 10.1073/pnas.91.23.11089
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Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator.

Abstract: The yeast two-hybrid system was used to identif proteins that interact with Ras. The H-Ras protein was found to interact with a g nnucleotide di ostimulator (GDS) that has been previously shown to regulate guanine nudeotide exchange on another member of the Ras protein family, Ral. The interaction is meted by the C-terminal, catalytic segment ofthe RalGDS and can be detected both in vivo, using the two-hybrid system, and in vto, with purfi recombinant proein. The inter of the RaIGDS C-teri segment with Ras is … Show more

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Cited by 272 publications
(207 citation statements)
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“…Cell lysates were incubated with glutathioneSepharose coupled peptides corresponding to RalGDS RBD (Hofer et al, 1994) or Raf-1 RBD (Kolch et al, 1996) as described in Materials and methods. Bound GTP-Ras was eluted from the beads and analysed by immunoblotting with anti-Ras (aRas) or anti-HA (aHA) antibodies.…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…Cell lysates were incubated with glutathioneSepharose coupled peptides corresponding to RalGDS RBD (Hofer et al, 1994) or Raf-1 RBD (Kolch et al, 1996) as described in Materials and methods. Bound GTP-Ras was eluted from the beads and analysed by immunoblotting with anti-Ras (aRas) or anti-HA (aHA) antibodies.…”
Section: Resultsmentioning
confidence: 99%
“…Following lysis in RIPA bu er (50 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.5% DOC, 1% NP40, 0.1% SDS, 25 mM NaF, 2 mM Na 3 VO 4 , aprotinin, leupeptin, pefabloc each 10 mg/ml), protein concentrations were determined using DC Protein Assay (BioRad Laboratories, Hercules, CA, USA). Total cell protein (400 mg) was incubated with glutathione-Sepharose coupled peptides corresponding to the Ras binding domains (RBDs) (5 mg/ml beads) of RalGDS (Hofer et al, 1994) or Raf-1 (Kolch et al, 1996) for 1 h at 48C. The beads were washed three times with RIPA bu er, bound proteins eluted in sample bu er, heated for 5 min at 958C, separated on 12% SDS ± PAGE, transferred to nitrocellulose and subjected to immunoblotting with anti-Ras or anti-HA antibodies.…”
Section: Ras Activationmentioning
confidence: 99%
“…Ras e ector molecules include the Raf kinases (Moodie et al, 1993Jaiswal et al, 1994;Van Aelst et al, 1993;Vojtek et al, 1993), MEKK (Russell et al, 1995), RalGDS family members (Hofer et al, 1994;Kikuchi et al, 1994;Spaargaren and Bischo , 1994;Wolthius et al, 1996), PI 3-OH kinase (PI3K) (Rodriguez-Viciana et al, 1994), Neuro®bromin (DiBattiste et al, 1993) and AF6 (Kuriyama et al, 1996). The mechanisms which determine and regulate the formation of speci®c Rase ector signalling complexes are not known.…”
Section: Introductionmentioning
confidence: 99%
“…Finally, the increasing number of candidate Ras e ectors provides additional evidence for the existence of Raf-independent Ras signaling pathways (Marshall, 1996;Khosravi-Far and Der, 1994). Included in this roster of functionally diverse proteins are the two Ras GTPase activating proteins (p120 and NF1 GAPs), three guanine nucleotide exchange factors of the Rasrelated proteins RalA and RalB (RalGDS, RGL, and Rlf/RGL2) (Kikuchi et al, 1994;Hofer et al, 1994;Spaargaren and Bischo , 1994;Wolthuis et al, 1996;Peterson et al, 1996), phosphatidylinositol-3-OH kinase (PI3K), (Rodriguez-Viciana et al, 1994), and Rin-1 (Han and Colicelli, 1995). Like Raf-1, these proteins show preferential binding to active Ras-GTP, and they require an intact Ras e ector domain (residues 32 ± 40) for this interaction.…”
Section: Introductionmentioning
confidence: 99%