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RLIP76, an effector of the GTPase Ral, interacts with the AP2 complex: involvement of the Ral pathway in receptor endocytosis
V. Jullien-Flores, Y. Mahe, G. Mirey, C. Leprince, B. Meunier-Bisceuil, A. Sorkin, J.H. Camonis
Journal of Cell Science 2000 113: 2837-2844;

Summary

RLIP76 is a modular protein that was identified as a putative effector of Ral, a GTPase activated during Ras signaling. To explore further the contribution of the Ral-RLIP76 pathway to Ras signaling, we have looked for partners of RLIP76. Mu2, the medium chain of the AP2 complex is shown to interact with RLIP76. We show also that in vivo endogenous AP2 and RLIP76 form a complex and that this in vivo interaction is independent of cells being stimulated by a growth factor. Furthermore, RLIP76 differentiates AP2 from AP1 in vivo as RLIP76 differentiates mu2 from mu1 in vitro and in two hybrid assays. We show that activated Ral interferes with both tranferrin receptor endocytosis and epidermal growth factor (EGF) receptor endocytosis in HeLa cells. We propose a model where the Ral-RLIP76 pathway connects signal transduction and endocytosis through interaction on one hand between the Ras-Ral pathway and RLIP, on the other hand between RLIP and proteins belonging to the endocytotic machinery.

REFERENCES

    1. Aguilar, R. C.,
    2. Ohno, H.,
    3. Roche, K. W. and
    4. Bonifacino, J. S.
    (1997). Functional domain mapping of the clathrin-associated adaptor medium chains mu1 and mu2. J. Biol. Chem 272, 27160–27166
    OpenUrlAbstract/FREE Full Text
    1. Albright, C. F.,
    2. Giddings, B. W.,
    3. Liu, J.,
    4. Vito, M. and
    5. Weinberg, R. A.
    (1993). Characterization of a guanine nucleotide dissociation stimulator for a ras-related GTPase. EMBO J 12, 339–347
    OpenUrlPubMedWeb of Science
    1. Bauer, B.,
    2. Mirey, G.,
    3. Vetter, I. R.,
    4. Garcia-Ranea, J. A.,
    5. Valencia, A.,
    6. Wittinghofer, A.,
    7. Camonis, J. H. and
    8. Cool, R. H.
    (1999). Effector recognition by the small GTP-binding proteins Ras and Ral. J. Biol Chem 274, 17763–17770
    OpenUrlAbstract/FREE Full Text
    1. Boll, W.,
    2. Ohno, H.,
    3. Songyang, Z.,
    4. Rapoport, I.,
    5. Cantley, L. C.,
    6. Bonifacino, J. S. and
    7. Kirchhausen, T.
    (1996). Sequence requirements for the recognition of tyrosine-based endocytic signals by clathrin AP-2 complexes. EMBO J 15, 5789–5795
    OpenUrlPubMedWeb of Science
    1. Bonifacino, J. S. and
    2. Dell'Angelica, E. C.
    ) (1999). Molecular bases for the recognition of tyrosine-based sorting signals. J. Cell Biol 145, 923–926
    OpenUrlFREE Full Text
    1. Cantor, S. B.,
    2. Urano, T. and
    3. Feig, L. A.
    (1995). Identification and characterization of Ral-binding protein 1, a potential downstream target of Ral GTPases. Mol. Cell. Biol 15, 4578–4584
    OpenUrlAbstract/FREE Full Text
    1. Cao, T. T.,
    2. Mays, R. W. and
    3. von Zastrow, M.
    (1998). Regulated endocytosis of G-protein-coupled receptors by a biochemically and functionally distinct subpopulation of clathrin-coated pits. J. Biol. Chem 273, 24592–24602
    OpenUrlAbstract/FREE Full Text
    1. Carter, L. L.,
    2. Redelmeier, T. E.,
    3. Woollenweber, L. A. and
    4. Schmid, S. L.
    (1993). Multiple GTP-binding proteins participate in clathrin-coated vesicle-mediated endocytosis. J. Cell Biol 120, 37–45
    OpenUrlAbstract/FREE Full Text
    1. Carter, R. E. and
    2. Sorkin, A.
    (1998). Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera. J. Biol. Chem 273, 35000–35007
    OpenUrlAbstract/FREE Full Text
    1. Gossen, M.,
    2. Freundlieb, S.,
    3. Bender, G.,
    4. Muller, G.,
    5. Hillen, W. and
    6. Bujard, H.
    (1995). Transcriptional activation by tetracyclines in mammalian cells. Science 268, 1766–1769
    OpenUrlAbstract/FREE Full Text
    1. Hofer, F.,
    2. Fields, S.,
    3. Schneider, C. and
    4. Martin, G. S.
    (1994). Activated Ras interacts with the Ral guanine nucleotide dissociation stimulator. Proc. Nat. Acad. Sci. USA 91, 11089–11093
    OpenUrlAbstract/FREE Full Text
    1. Ikeda, M.,
    2. Ishida, O.,
    3. Hinoi, T.,
    4. Kishida, S. and
    5. Kikuchi, A.
    (1998). Identification and characterization of a novel protein interacting with Ral-binding protein 1, a putative effector protein of Ral. J. Biol. Chem 273, 814–821
    OpenUrlAbstract/FREE Full Text
    1. Jullien-Flores, V.,
    2. Dorseuil, O.,
    3. Romero, F.,
    4. Letourneur, F.,
    5. Saragosti, S.,
    6. Berger, R.,
    7. Tavitian, A.,
    8. Gacon, G. and
    9. Camonis, J.
    (1995). Bridging theRal GTPase to Rho pathways: RLIP76, a Ral effector with CDC42/Rac GTPase activating protein (GAP) activity. J. Biol. Chem 270, 22473–22477
    OpenUrlAbstract/FREE Full Text
    1. Kikuchi, A.,
    2. Demo, S. D.,
    3. Ye, Z.-H.,
    4. Chen, Y.-W. and
    5. Williams, L. T.
    (1994). RalGDS family members interact with the effector loop of ras p21. Mol. Cell. Biol 14, 7483–7491
    OpenUrlAbstract/FREE Full Text
    1. Kim, J. H.,
    2. Lee, S. D.,
    3. Han, J. M.,
    4. Lee, T. G.,
    5. Kim, Y.,
    6. Park, J. B.,
    7. Lambeth, J. D.,
    8. Suh, P. G. and
    9. Ryu, S. H.
    (1998). Activation of phospholipase D1 by direct interaction with ADP-ribosylation factor 1 and RalA. FEBS Lett 430, 231–235
    OpenUrlCrossRefPubMedWeb of Science
    1. L'Allemain, G.,
    2. Seuwen, K.,
    3. Velu, T. and
    4. Pouyssegur, J.
    (1989). Signal transduction in hamster fibroblasts overexpressing the human EGF receptor. Growth Factors 1, 311–321
    OpenUrlCrossRefPubMed
    1. Lopez-Barahona, M.,
    2. Bustelo, X. R. and
    3. Barbacid, M.
    (1996). The TC21 oncoprotein interacts with the Ral guanosine nucleotide dissociation factor. Oncogene 12, 463–470
    OpenUrlPubMed
    1. Matsubara, K.,
    2. Hinoi, T.,
    3. Koyama, S. and
    4. Kikuchi, A.
    (1998). Post-translational modifications of Ral and Rac1 are important for the action of Ral-binding protein 1, a putative effector protein of Ral. FEBS Lett 410, 169–174
    OpenUrl
    1. Nakashima, S.,
    2. Morinaka, K.,
    3. Koyama, S.,
    4. Ikeda, M.,
    5. Kishida, M.,
    6. Okawa, K.,
    7. Iwamatsu, A.,
    8. Kishida, S. and
    9. Kikuchi, A.
    (1999). Small G protein Ral and its downstream molecules regulate endocytosis of EGF and insulin receptors. EMBO J 18, 3629–3642
    OpenUrlAbstract
    1. Nesterov, A.,
    2. Carter, R. E.,
    3. Sorkina, T.,
    4. Gill, G. N. and
    5. Sorkin, A.
    (1999). Inhibition of the receptor-binding function of clathrin adaptor protein AP-2 by dominant-negative mutant mu2 subunit and its effects on endocytosis. EMBO J 18, 2489–2499
    OpenUrlAbstract
    1. Ohno, H.,
    2. Stewart, J.,
    3. Fournier, M. C.,
    4. Bosshart, H.,
    5. Rhee, I.,
    6. Miyatake, S.,
    7. Saito, T.,
    8. Gallusser, A.,
    9. Kirchhausen, T. and
    10. Bonifacino, J. S.
    (1995). Interaction of tyrosine-based sorting signals with clathrin-associated proteins. Science 269, 1872–1875
    OpenUrlAbstract/FREE Full Text
    1. Park, S.-H. and
    2. Weinberg, R. A.
    (1996). A putative effector of Ral has homology to Rho/Rac GTPase activating proteins. Oncogene 11, 2349–2355
    OpenUrl
    1. Schmid, S. L.,
    2. McNiven, M. A. and
    3. De Camilli, P.
    (1998). Dynamin and its partners: a progress report. Curr. Opin. Cell. Biol 10, 504–512
    OpenUrlCrossRefPubMedWeb of Science
    1. Spaargaren, M. and
    2. Bischoff, J. R.
    (1994). Identification of the guanine nucleotide dissociation stimulator for Ral as a putative effector molecule of R-ras, H-ras, K-ras and Rap. Proc. Nat. Acad. Sci. USA 91, 12609–12613
    OpenUrlAbstract/FREE Full Text
    1. Urano, T.,
    2. Emkey, R. and
    3. Feig, L. A.
    (1996). Ral-GTPases mediate a distinct downstream signaling pathway from Ras that facilitates cellular transformation. EMBO J 15, 810–816
    OpenUrlPubMedWeb of Science
    1. Vojtek, A. B.,
    2. Hollenberg, S. M. and
    3. Cooper, J. A.
    (1993). Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell 74, 205–214
    OpenUrlCrossRefPubMedWeb of Science
    1. White, M. A.,
    2. Vale, T.,
    3. Camonis, J. H.,
    4. Schaefer, E. and
    5. Wigler, M. H.
    (1996). A role for Ral guanine nucleotide dissociation stimulator in mediating Ras-induced transformation. J. Biol. Chem 271, 16439–16442
    OpenUrlAbstract/FREE Full Text
    1. Wilde, A.,
    2. Beattie, E. C.,
    3. Lem, L.,
    4. Riethof, D. A.,
    5. Liu, S. H.,
    6. Mobley, W. C.,
    7. Soriano, P. and
    8. Brodsky, F. M.
    (1999). EGF receptor signaling stimulates SRC kinase phosphorylation of clathrin, influencing clathrin redistribution and EGF uptake. Cell 96, 677–687
    OpenUrlCrossRefPubMedWeb of Science
    1. Wolthuis, R. M.,
    2. Bauer, B.,
    3. van't Veer, L. J.,
    4. de Vries-Smits, A. M.,
    5. Cool, R. H.,
    6. Spaargaren, M.,
    7. Wittinghofer, A.,
    8. Burgering, B. M. and
    9. Bos, J. L.
    (1996). RalGDS-like factor (Rlf) is a novel Ras and Rap 1A-associating protein. Oncogene 13, 353–362
    OpenUrlPubMedWeb of Science
    1. Wolthuis, R. M.,
    2. de Ruiter, N. D.,
    3. Cool, R. H. and
    4. Bos, J. L.
    (1997). Stimulation of gene induction and cell growth by the Ras effector Rlf. EMBO J 16, 6748–6761
    OpenUrlAbstract
    1. Wolthuis, R. M.,
    2. Franke, B.,
    3. van Triest, M.,
    4. Bauer, B.,
    5. Cool, R. H.,
    6. Camonis, J. H.,
    7. Akkerman, J. W. and
    8. Bos, J. L.
    (1998). Activation of the small GTPase Ral in platelets. Mol. Cell. Biol 18, 2486–2491
    OpenUrlAbstract/FREE Full Text
    1. Wolthuis, R. M.,
    2. Zwartkruis, F.,
    3. Moen, T. C. and
    4. Bos, J. L.
    (1998). Ras-dependent activation of the small GTPase Ral. Curr. Biol 8, 471–474
    OpenUrlCrossRefPubMedWeb of Science
    1. Yamaguchi, A.,
    2. Urano, T.,
    3. Goi, T. and
    4. Feig, L. A.
    (1997). An Eps homology (EH) domain protein that binds to the Ral-GTPase target, RalBP1. J. Biol. Chem 272, 31230–31234
    OpenUrlAbstract/FREE Full Text
    1. Zhang, Y. Q. and
    2. Broadie, K.
    (1999). Cloning, mapping and tissue-specific expression of Drosophila clathrin-associated protein AP50 gene. Gene 233, 171–179
    OpenUrlCrossRefPubMedWeb of Science
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Journal Article
RLIP76, an effector of the GTPase Ral, interacts with the AP2 complex: involvement of the Ral pathway in receptor endocytosis
V. Jullien-Flores, Y. Mahe, G. Mirey, C. Leprince, B. Meunier-Bisceuil, A. Sorkin, J.H. Camonis
Journal of Cell Science 2000 113: 2837-2844;
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