A large percentage of current drugs target G-protein-coupled receptors, which couple to well-known signaling pathways involving cAMP or calcium. G-proteins themselves may subserve a second messenger function. Here, we review the role of tubulin and microtubules in directly mediating effects of heterotrimeric G-proteins on neuronal outgrowth, shape and differentiation. G-protein-tubulin interactions appear to be regulated by neurotransmitter activity, and, in turn, regulate the location of Gα in membrane microdomains (such as lipid rafts) or the cytosol. Tubulin binds with nanomolar affinity to Gsα, Giα1 and Gqα (but not other Gα subunits) as well as Gβ1γ2 subunits. Gα subunits destabilize microtubules by stimulating tubulin’s GTPase, while Gβγ subunits promote microtubule stability. The same region on Gsα that binds adenylyl cyclase and Gβγ also interacts with tubulin, suggesting that cytoskeletal proteins are novel Gα effectors. Additionally, intracellular Giα-GDP, in concert with other GTPase proteins and Gβγ, regulates the position of the mitotic spindle in mitosis. Thus, G-protein activation modulates cell growth and differentiation by directly altering microtubule stability. Further studies are needed to fully establish a structural mechanism of this interaction and its role in synaptic plasticity.
© 2009 S. Karger AG, Basel
- Lipid rafts
- Microtubule-associated protein
- Neurite outgrowth
- Adam RM, Yang W, Di Vizio D, Mukhopadhyay NK, Steen H: Rapid preparation of nuclei-depleted detergent-resistant membrane fractions suitable for proteomics analysis. BMC Cell Biol 2008;9:30.
- Allen JA, Halverson-Tamboli RA, Rasenick MM: Lipid raft microdomains and neurotransmitter signalling. Nat Rev Neurosci 2007;8:128–140.
- Allen JA, Yu JZ, Donati RJ, Rasenick MM: β-Adrenergic receptor stimulation promotes Gαs internalization through lipid rafts: a study in living cells. Mol Pharmacol 2005;67:1493–1504.
- Arthur DB, Akassoglou K, Insel PA: P2Y2 and TrkA receptors interact with Src family kinase for neuronal differentiation. Biochem Biophys Res Commun 2006;347:678–682.
- Arthur DB, Akassoglou K, Insel PA: P2Y2 receptor activates nerve growth factor/TrkA signaling to enhance neuronal differentiation. Proc Natl Acad Sci USA 2005;102:19138–19143.
- Arvanitis DN, Min W, Gong Y, Heng YM, Boggs JM: Two types of detergent-insoluble, glycosphingolipid/cholesterol-rich membrane domains from isolated myelin. J Neurochem 2005;94:1696–1710.
- Bünemann M, Frank M, Lohse MJ: Gi protein activation in intact cells involves subunit rearrangement rather than dissociation. Proc Natl Acad Sci USA 2003;100:16077–16082.
- Burnette DT, Schaefer AW, Ji L, Danuser G, Forscher P: Filopodial actin bundles are not necessary for microtubule advance into the peripheral domain of aplysia neuronal growth cones. Nat Cell Biol 2007;9:1360–1369.
- Calvert PD, Strissel KJ, Schiesser WE, Pugh EN Jr, Arshavsky VY: Light-driven translocation of signaling proteins in vertebrate photoreceptors. Trends Cell Biol 2006;16:560–568.
- Cazillis M, Gonzalez BJ, Billardon C, Lombet A, Fraichard A, Samarut J, Gressens P, Vaudry H, Rostène W: VIP and PACAP induce selective neuronal differentiation of mouse embryonic stem cells. Eur J Neurosci 2004;19:798–808.
- Chen NF, Yu JZ, Skiba NP, Hamm HE, Rasenick MM: A specific domain of Giα required for the transactivation of Giα by tubulin is implicated in the organization of cellular microtubules. J Biol Chem 2003;278:15285–15290.
- Ciruela F, Robbins MJ, Willis AC, McIlhinney RA: Interactions of the C-terminus of metabotropic glutamate receptor type 1α with rat brain proteins: evidence for a direct interaction with tubulin. J Neurochem 1999;72:346–354.
- David-Pfeuty T, Laporte J, Pantaloni D: GTPase activity at ends of microtubules. Nature 1978;272:282–284.
- David-Pfeuty T, Simon C, Pantaloni D: Effect of antimitotic drugs on tubulin GTPase activity and self-assembly. J Biol Chem 1979;254:11696–11702.
- Davis A, Sage CR, Dougherty CA, Farrell KW: Microtubule dynamics modulated by guanosine triphosphate hydrolysis activity of β-tubulin. Science 1994;264:839–842.
- Donati RJ, Rasenick MM: Chronic antidepressant treatment prevents accumulation of Gs in cholesterol-rich, cytoskeletal-associated plasma membrane domains (lipid rafts). Neuropsychopharmacology 2005;30:1238–1245.
- Fleury D, Grenningloh G, Lafanechene L, Antonsson B, Job D, Cohen-Addad C: Preliminary crystallographic study of a complex formed between the α/β-tubulin heterodimer and the neuronal growth-associated protein SCG10. J Struct Biol 2000;131:156–158.
- Foster LJ, de Hoog CL, Mann M: Unbiased quantitative proteomics of lipid rafts reveals high specificity for signaling factors. Proc Natl Acad Sci USA 2003;100:5813–5818.
- Gigant B, Wang C, Ravelli RB, Roussi F, Steinmetz MO, Curmi PA, Sobel A, Knossow M: Structural basis for the regulation of tubulin by vinblastine. Nature 2005;435:519–522.
- Gotta M, Ahringer J: Axis determination in C. elegans: initiating and transducing polarity. Curr Opin Genet Dev 2001;11:367–373.
- He JC, Neves SR, Jordan JD, Iyengar R: Role of the Go/i signaling network in the regulation of neurite outgrowth. Can J Physiol Pharmacol 2006;84:687–694.
- Head BP, Patel HH, Roth DM, Murray F, Swaney JS, Niesman IR, Farquar MG, Insel PA: Microtubules and actin microfilaments regulate lipid raft/caveolae localization of adenylyl cyclase signaling components. J Biol Chem 2006;281:26391–26399.
- Hynes TR, Mervine SM, Yost EA, Sabo JL, Berlot CH: Live cell imaging of Gs and the β2-adrenergic receptor demonstrates that both αs and β1γ7 internalize upon stimulation and exhibit similar trafficking patterns that differ from that of the β2-adrenergic receptor. J Biol Chem 2004;279:44101–44112.
- Hynes TR, Tang L, Mervine SM, Sabo JL, Yost EA, Devreotes PN, Berlot CH: Visualization of G-protein βγ dimers using bimolecular fluorescence complementation demonstrates roles for both β and γ in subcellular targeting. J Biol Chem 2004;279:30279–30286.
- Igarashi M, Strittmatter SM, Vartanian T, Fishman MC: Mediation by G-proteins of signals that cause collapse of growth cones. Science 1993;259:77–79.
- Janich P, Corbeil D: GM1 and GM3 gangliosides highlight distinct lipid microdomains within the apical domain of epithelial cells. FEBS Lett 2007;581:1783–1787.
- Jarzynka MJ, Passey DK, Ignatius PF, Melan MA, Radio NM, Jockers R, Rasenick MM, Brydon L, Witt-Enderby PA: Modulation of melatonin receptors and G-protein function by microtubules. J Pineal Res 2006;41:324–336.
- Jolly C, Mitar I, Sattentau QJ: Requirement for an intact T-cell actin and tubulin cytoskeleton for efficient assembly and spread of human immunodeficiency virus type 1. J Virol 2007;81:5547–5560.
- Kennedy MS, Insel PA: Inhibitors of microtubule assembly enhance β-adrenergic and prostaglandin E1-stimulated cyclic AMP accumulation in S49 lymphoma cells. Mol Pharmacol 1979;16:215–223.
- Lallemand-Breitenbach V, Quesnoit M, Braun V, El Marjou A, Pous C, Goud B, Perez F: CLIPR-59 is a lipid raft-associated protein containing a cytoskeleton-associated protein glycine-rich domain (CAP-Gly) that perturbs microtubule dynamics. J Biol Chem 2004;279:41168–41178.
- Layden BT, Saengsawang W, Donati RJ, Yang S, Mulhearn DC, Johnson ME, Rasenick MM: Structural model of a complex between the heterotrimeric G-protein, Gsα, and tubulin. Biochim Biophys Acta 2008;1783:964–973.
- Li N, Shaw AR, Zhang N, Mak A, Li L: Lipid raft proteomics: analysis of in-solution digest of sodium dodecyl sulfate-solubilized lipid raft proteins by liquid chromatography-matrix-assisted laser desorption/ionization tandem mass spectrometry. Proteomics 2004;4:3156–3166.
- Maekawa S, Morii H, Kumanogoh H, Sano M, Naruse Y, Sokawa Y, Mori N: Localization of neuronal growth-associated, microtubule-destabilizing factor SCG10 in brain-derived raft membrane microdomains. J Biochem 2001;129:691–697.
- Manna T, Grenningloh G, Miller HP, Wilson L: Stathmin family protein SCG10 differentially regulates the plus and minus end dynamics of microtubules at steady state in vitro: implications for its role in neurite outgrowth. Biochemistry 2007;46:3543–3552.
- Marta CB, Montano MB, Taylor CM, Taylor AL, Bansal R, Pfeiffer SE: Signaling cascades activated upon antibody cross-linking of myelin oligodendrocyte glycoprotein: potential implications for multiple sclerosis J Biol Chem 2005;280:8985–8993.
- Marta CB, Taylor CM, Coetzee T, Kim T, Winkler S, Bansal R, Pfeiffer SE: Antibody cross-linking of myelin oligodendrocyte glycoprotein leads to its rapid repartitioning into detergent-insoluble fractions, and altered protein phosphorylation and cell morphology. J Neurosci 2003;23:5461–5471.
- Miura Y, Hanada K, Jones TL: G(s) signaling is intact after disruption of lipid rafts. Biochemistry 2001;40:15418–15423.
- Montoya V, Gutierrez C, Najera O, Leony D, Varela-Ramirez A, Popova J, Rasenick MM, Das S, Roychowdhury S: G-protein βγ subunits interact with αβ- and γ-tubulin and play a role in microtubule assembly in PC12 cells. Cell Motil Cytoskeleton 2007;64:936–950.
- Morii H, Shiraishi-Yamaguchi Y, Mori N: SCG10, a microtubule-destabilizing factor, stimulates the neurite outgrowth by modulating microtubule dynamics in rat hippocampal primary cultured neurons. J Neurobiol 2006;66:1101–1114.
- Popova JS, Rasenick MM: Gβγ mediates the interplay between tubulin dimers and microtubules in the modulation of Gq signaling. J Biol Chem 2003;278:34299–34308.
- Rasenick MM, Stein PJ, Bitensky MW: The regulatory subunit of adenylate cyclase interacts with cytoskeletal components. Nature 1981;294:560–562.
- Rasenick MM, Donati RJ, Popova JS, Yu J-Z: Tubulin as a regulator of G-protein signaling. Meth Enzymol 2004;390:389–403.
- Ravelli RB, Gigant B, Curmi PA, Jourdain I, Lachkar S, Sobel A, Knossow M: Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain. Nature 2004;428:198–202.
- Reidel B, Goldmann T, Giessl A, Wolfrum U: The translocation of signaling molecules in dark adapting mammalian rod photoreceptor cells is dependent on the cytoskeleton. Cell Motil Cytoskeleton 2008;65:785–800.
- Riederer BM, Pellier V, Antonsson B, Di Paolo G, Stimpson SA, Lutjens R, Catsicas S, Grenningloh G: Regulation of microtubule dynamics by the neuronal growth-associated protein SCG10. Proc Natl Acad Sci USA 1997;94:741–745.
- Roychowdhury S, Martinez L, Salgado L, Das S, Rasenick MM: G-protein activation is prerequisite for functional coupling between Gα/Gβγ and tubulin/microtubules. Biochem Biophys Res Commun 2006;340:441–448.
- Roychowdhury S, Panda D, Wilson L, Rasenick MM: G-protein α subunits activate tubulin GTPase and modulate microtubule polymerization dynamics. J Biol Chem 1999;274:13485–13490.
- Roychowdhury S, Rasenick MM: G-protein β1γ2 subunits promote microtubule assembly. J Biol Chem 1997;272:31576–31581.
- Roychowdhury S, Rasenick MM: Submembraneous microtubule cytoskeleton: regulation of microtubule assembly by heterotrimeric G-proteins. FEBS J 2008;275:4654– 4663.
- Rudolph SA, Hegstrand LR, Greengard P, Malawista SE: The interaction of colchicine with hormone-sensitive adenylate cyclase in human leukocytes. Mol Pharmacol 1979;16:805–812.
- Rybin VO, Xu X, Lisanti MP, Steinberg SF: Differential targeting of β-adrenergic receptor subtypes and adenylyl cyclase to cardiomyocyte caveolae. A mechanism to functionally regulate the cAMP signaling pathway. J Biol Chem 2000;275:41447–41457.
- Sarma T, Voyno-Yasenetskaya T, Hope TJ, Rasenick MM: Heterotrimeric G-proteins associate with microtubules during differentiation in PC12 pheochromocytoma cells. FASEB J 2003;17:848–859.
- Sato M, Cismowski MJ, Toyota E, Smrcka AV, Lucchesi PA, Chilian WM, Lanier SM: Identification of a receptor-independent activator of G-protein signaling (AGS8) in ischemic heart and its interaction with Gβγ. Proc Natl Acad Sci USA 2006;103:797–802.
- Schaefer M, Petronczki M, Dorner D, Forte M, Knoblich JA: Heterotrimeric G-proteins direct two modes of asymmetric cell division in the Drosophila nervous system. Cell 2001;107:183–194.
- Siller KH, Cabernard C, Doe CQ: The NuMA-related MUD protein binds PINS and regulates spindle orientation in Drosophila neuroblasts. Nat Cell Biol 2006;8:594–600.
- Simons K, Toomre D: Lipid rafts and signal transduction. Nat Rev Mol Cell Biol 2000;1:31–39.
- Slepak VZ, Hurley JB: Mechanism of light-induced translocation of arrestin and transducin in photoreceptors: interaction-restricted diffusion. IUBMB Life 2008;60:2–9.
- Stepanova T, Slemmer J, Hoogenraad CC, Lansbergen G, Dortland B, De Zeeuw CI, Grosveld F, van Cappellen G, Akhmanova A, Galjart N: Visualization of microtubule growth in cultured neurons via the use of EB3-GFP (end-binding protein 3-green fluorescent protein). J Neurosci 2003;23:2655–2664.
- Thomas CJ, Tall GG, Adhikari A, Sprang SR: Ric-8A catalyzes guanine nucleotide exchange on Gαi1 bound to the GPR/GoLoco exchange inhibitor AGS3. J Biol Chem 2008;283:23150–23160.
- Wang N, Yan K, Rasenick MM: Tubulin binds specifically to the signal-transducing proteins, Gsα and Giα1. J Biol Chem 1990;265:1239–1242.
- Willard FS, Kimple RJ, Siderovski DP: Return of the GDI: the GoLoco motif in cell division. Annu Rev Biochem 2004;73:925–951.
- Xiao H, Verdier-Pinard P, Fernandez-Fuentes N, Burd B, Angeletti R, Fiser A, Horwitz SB, Orr GA: Insights into the mechanism of microtubule stabilization by taxol. Proc Natl Acad Sci USA 2006;103:10166–10173.
- Yan K, Green E, Belga F, Rasenick MM: Synaptic membrane G-proteins are complexed with tubulin in situ. J Neurochem 1996;66:1489–1495.
- Yan K, Popova JS, Moss A, Shah B, Rasenick MM: Tubulin stimulates adenylyl cyclase activity in C6 glioma cells by bypassing the β-adrenergic receptor: a potential mechanism of G-protein activation. J Neurochem 2001;76:182–190.
- Yu H, Wakim B, Li M, Halligan B, Tint S, Patel SB: Quantifying raft proteins in neonatal mouse brain by ‘tube-gel’ protein digestion label-free shotgun proteomics. Proteome Sci 2007;5:17.
- Yu JZ, Rasenick MM: Real-time visualization of a fluorescent Gαs: dissociation of the activated G-protein from plasma membrane. Mol Pharmacol 2002;61:352–359.
- Van Rossum D, Hanisch UK: Cytoskeletal dynamics in dendritic spines: direct modulation by glutamate receptors? Trends Neurosci 1999;22:290–295.
- Popova JS, Rasenick MM: Clathrin-mediated endocytosis of M3 muscarinic receptors. Roles for Gβγ and tubulin. J Biol Chem 2004;279:30410–30418.
- Jordan BA, Trapaidze N, Gomes I, Nivarthi R, Devi LA: Oligomerization of opioid receptors with β2-adrenergic receptors: a role in trafficking and mitogen-activated protein kinase activation. Proc Natl Acad Sci USA 2001;98:343–348.
- Daaka Y, Luttrell LM, Lefkowitz RJ: Switching of the coupling of the β2-adrenergic receptor to different G-proteins by protein kinase A: Nature 1997;390:88–91.
- McGraw DW, Elwing JM, Fogel KM, Wang WC, Glinka CB, Mihlbachler KA, Rothenberg ME, Liggett SB: Crosstalk between Gi and Gq/Gs pathways in airway smooth muscle regulates bronchial contractility and relaxation. J Clin Invest 2007;117:1391–1398.
- Popova JS, Rasenick MM: Muscarinic receptor activation promotes the membrane association of tubulin for the regulation of Gq-mediated phospholipase Cβ1 signaling. J Neurosci 2000;20:2774–2782.
- Popova JS, Greene AK, Wang J, Rasenick MM: Phosphatidylinositol 4,5-bisphosphate modifies tubulin participation in phospholipase Cβ1 signaling. J Neurosci 2002;22:1668–1678.
- Ciruela F, McIlhinney RA: Metabotropic glutamate receptor type 1α and tubulin assemble into dynamic interacting complexes. J Neurochem 2001;76:750–758.
- Du Q, Macara IG: Mammalian Pins is a conformational switch that links NuMA to heterotrimeric G-proteins. Cell 2004;119:503–516.
- Chen R, Tong W, Mintseris J, Li L, Weng Z: ZDOCK predictions for the CAPRI challenge. Proteins 2003;52:68–73.
- Comeau SR, Gatchell DW, Vajda S, Camacho CJ: ClusPro: a fully automated algorithm for protein-protein docking. Nucleic Acids Res 2004;32:W96–W99.
- Sunahara RK, Tesmer JJ, Gilman AG, Sprang SR: Crystal structure of the adenylyl cyclase activator Gsα. Science 1997;278:1943–1947.
Mark M. Rasenick
Departments of Physiology and Biophysics, Psychiatry, and
Graduate Program in Neuroscience, University of Illinois
Chicago, IL 60612-7342 (USA)
Tel. +1 312 996 6641, Fax +1 312 996 1414, E-Mail firstname.lastname@example.org
Received: September 19, 2008
Accepted after revision: November 5, 2008
Published online: February 12, 2009
Number of Print Pages : 9
Number of Figures : 2, Number of Tables : 0, Number of References : 78
Vol. 17, No. 1, Year 2009 (Cover Date: February 2009)
Journal Editor: Ip N.Y. (Hong Kong)
ISSN: 1424-862X (Print), eISSN: 1424-8638 (Online)
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