Gβγ Mediates the Interplay between Tubulin Dimers and Microtubules in the Modulation of Gq Signaling

Agonist stimulation causes tubulin association with the plasma membrane and activation of PLCβ1 through direct interaction with, and transactivation of, Gαq. Here we demonstrate that Gβγ interaction with tubulin down-regulates this signaling pathway. Purified Gβγ, alone or with phosphatidylinositol 4,5-bisphosphate (PIP2), inhibited carbachol-evoked membrane recruitment of tubulin and Gαq transactivation by tubulin. Polymerization of microtubules elicited by Gβγ overrode tubulin translocation to the membrane in response to carbachol stimulation. Gβγ sequestration of tubulin reduced the inhibition of PLCβ1 observed at high tubulin concentration. Gβ1γ2 interacted preferentially with tubulin-GDP, whereas Gαq was transactivated by tubulin-GTP. Prenylation of the γ2 polypeptide was required for Gβγ/tubulin interaction. Both confocal microscopy and coimmunoprecipitation studies revealed the spatiotemporal pattern of Gβγ/tubulin interaction during carbachol stimulation of neuroblastoma SK-N-SH cells. In resting cells Gβγ localized predominantly at the cell membrane, whereas tubulin was found in well defined microtubules in the cytosol. Within 2 min of agonist exposure, a subset of tubulin translocated to the plasma membrane and colocalized with Gβ. Fifteen min post-carbachol addition, tubulin and Gβ colocalized in vesicle-like structures in the cytosol. Gβ/tubulin colocalization increased after pretreatment of cells with the microtubule-depolymerizing agent, colchicine, and was inhibited by taxol. Taxol also inhibited carbachol-induced PIP2 hydrolysis. It is suggested that Gβγ/tubulin interaction mediates internalization of membrane-associated tubulin at the offset of PLCβ1 signaling. Newly cytosolic Gβγ/tubulin complexes might promote microtubule polymerization attenuating further tubulin association with the plasma membrane. Thus G protein-coupled receptors might evoke Gα and Gβγ to orchestrate regulation of phospholipase signaling by tubulin dimers and control of cell shape by microtubules. Agonist stimulation causes tubulin association with the plasma membrane and activation of PLCβ1 through direct interaction with, and transactivation of, Gαq. Here we demonstrate that Gβγ interaction with tubulin down-regulates this signaling pathway. Purified Gβγ, alone or with phosphatidylinositol 4,5-bisphosphate (PIP2), inhibited carbachol-evoked membrane recruitment of tubulin and Gαq transactivation by tubulin. Polymerization of microtubules elicited by Gβγ overrode tubulin translocation to the membrane in response to carbachol stimulation. Gβγ sequestration of tubulin reduced the inhibition of PLCβ1 observed at high tubulin concentration. Gβ1γ2 interacted preferentially with tubulin-GDP, whereas Gαq was transactivated by tubulin-GTP. Prenylation of the γ2 polypeptide was required for Gβγ/tubulin interaction. Both confocal microscopy and coimmunoprecipitation studies revealed the spatiotemporal pattern of Gβγ/tubulin interaction during carbachol stimulation of neuroblastoma SK-N-SH cells. In resting cells Gβγ localized predominantly at the cell membrane, whereas tubulin was found in well defined microtubules in the cytosol. Within 2 min of agonist exposure, a subset of tubulin translocated to the plasma membrane and colocalized with Gβ. Fifteen min post-carbachol addition, tubulin and Gβ colocalized in vesicle-like structures in the cytosol. Gβ/tubulin colocalization increased after pretreatment of cells with the microtubule-depolymerizing agent, colchicine, and was inhibited by taxol. Taxol also inhibited carbachol-induced PIP2 hydrolysis. It is suggested that Gβγ/tubulin interaction mediates internalization of membrane-associated tubulin at the offset of PLCβ1 signaling. Newly cytosolic Gβγ/tubulin complexes might promote microtubule polymerization attenuating further tubulin association with the plasma membrane. Thus G protein-coupled receptors might evoke Gα and Gβγ to orchestrate regulation of phospholipase signaling by tubulin dimers and control of cell shape by microtubules. Binding of agonists to G protein-coupled receptors causes receptors to interact with specific G protein α-βγ heterotrimers, which, in turn, triggers the replacement of GDP for GTP on the α subunit and the functional dissociation and activation of both α and βγ subunits. Gα and Gβγ are then able to activate effectors and initiate signal transduction. Tubulin is a cytoskeletal protein that forms microtubules (1Desai A. Mitchison T.J. Annu. Rev. Cell Dev. Biol. 1997; 13: 83-117Crossref PubMed Scopus (1978) Google Scholar, 2Stewart R.J. Farrell K.W. Wilson L. Biochemistry. 1990; 29: 6489-6498Crossref PubMed Scopus (68) Google Scholar) and regulates G protein-mediated signaling (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar, 4Roychowdhury S. Rasenick M.M. Biochemistry. 1994; 33: 9800-9805Crossref PubMed Scopus (71) Google Scholar, 5Wang N. Yan K. Rasenick M.M. J. Biol. Chem. 1990; 265: 1239-1242Abstract Full Text PDF PubMed Google Scholar) through binding and hydrolysis of GTP. Transfer of GTP from tubulin to the α subunit of Gi1, Gs, and Gq (transactivation) leads to Gα coupling to, and regulation of, adenylyl cyclase and phospholipase Cβ1 (PLCβ1) 1The abbreviations used are: PLCβ1, phospholipase Cβ1; AAGTP, P3(4-azidoanilido)-P1-5′-GTP; tubulin-AAGTP, dimeric tubulin with AAGTP bound; GDPβS, guanyl-5′-yl thiophosphate; HPLC, high-performance liquid chromatography; Pipes, 1,4-piperazinediethanesulfonic acid; DTT, dithiothreitol; CHAPS, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; QNB, l-quinuclidinyl [phenyl-4(n)]benzilate, muscarinic receptor antagonist; PIP2, phosphatidylinositol 4,5-bisphosphate; IP3, inositol 1,4,5-trisphosphate; GRK2, G protein receptor kinase 2; DMEM, Dulbecco's modified Eagle's medium; PBS, phosphate-buffered saline; GTPγS, guanosine 5′-3-O-(thio)triphosphate; protein S. Rasenick M.M. Biochemistry. 1994; 33: 9800-9805Crossref PubMed Scopus (71) Google Scholar, 5Wang N. Yan K. Rasenick M.M. J. Biol. Chem. 1990; 265: 1239-1242Abstract Full Text PDF PubMed Google Scholar, Cell Biol. PubMed Scopus Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1994; Full Text PDF PubMed Google Tubulin to the plasma membrane to G by muscarinic receptors (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar, J.S. J. Rasenick M.M. J. Neurosci. PubMed Google Scholar) or receptors J. PubMed Scopus Google Scholar) causes microtubule and association of tubulin with plasma membrane in cells. binding to the plasma membrane tubulin which, in turn, PLCβ1 J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google of tubulin at the membrane PLCβ1 that tubulin to the PLCβ1 phosphatidylinositol J.S. J. Rasenick M.M. J. Neurosci. PubMed Google the of PIP2 inositol that microtubule PubMed Scopus Google inhibition of PLCβ1 by increased of tubulin dimers in to the plasma membrane J.S. J. Rasenick M.M. J. Neurosci. PubMed Google Both G protein α and βγ the of the microtubule in Gα tubulin and microtubule S. Wilson L. Rasenick M.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Gβγ microtubule polymerization and microtubules in S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google of to this the is and is microtubule this Gβγ on in microtubule the of a interaction in the cell and on signaling of Gβγ and PIP2 with the G protein receptor kinase 2 membrane translocation and the of the receptor K. R.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Gβγ might in translocation of tubulin to membrane-associated signaling of tubulin with Gα or Gβγ might tubulin microtubule polymerization in response to a Gβγ might cytosolic tubulin and tubulin in the regulation of signaling. the interaction of tubulin with Gβγ and the of this interaction on the PLCβ1 signaling and microtubule It is suggested that agonist stimulation tubulin to the membrane, Gαq. to PLCβ1 tubulin with Gβγ, and this to the cytosol. In membrane polymerization that complexes might microtubule from coimmunoprecipitation and in well from confocal microscopy or coimmunoprecipitation in a direct functional G protein-coupled receptor signaling and the of the microtubule direct for and G protein signaling and the of the microtubule a in the of cytoskeletal or to cell and cell Cell cells in J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google SK-N-SH neuroblastoma cells in Dulbecco's modified Eagle's with to J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google in cells with muscarinic PLCβ1 J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google of was S. Biochemistry. 1994; 33: PubMed Scopus Google Scholar, J. Biol. Chem. Full Text PDF PubMed Google Scholar, K. J. Biol. Chem. Full Text PDF PubMed Google from cells and or SK-N-SH cells in DTT, and membrane J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google was by the M.M. PubMed Scopus Google Scholar) with a of G and PLCβ1, was by to with specific for the muscarinic receptor from from or PLCβ1 from S. at a of or used by of the protein by of the by for a binding studies a to muscarinic receptor J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google with Gαq and PLCβ1 in the muscarinic receptor was at of membrane protein J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google SK-N-SH cells a high of muscarinic receptors of membrane to 33: Google Scholar, J. PubMed Scopus Google Scholar, Google of S. A. PubMed Scopus Google by and the tubulin was N. Rasenick M.M. Biochemistry. PubMed Scopus Google was and in liquid of GTP in was N. Yan K. Rasenick M.M. J. Biol. Chem. 1990; 265: 1239-1242Abstract Full Text PDF PubMed Google GTP was from by pretreatment by of tubulin with or for min on to through to the of of tubulin used the on the protein concentration. from cell was by J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google from was a from was by J. and A. Purified Gβγ also by Gβγ for the used in the 2 through a to S. N. Rasenick M.M. Biochemistry. PubMed Scopus Google was with SK-N-SH Gβγ both in the and of carbachol in polymerization Pipes, 2 Gβγ was with tubulin for min on was for min at in a at for min at was by of the and the in of polymerization on for and at for min at to from tubulin to and with a and in a carbachol on tubulin polymerization in the of SK-N-SH with with the of in the or of carbachol Gβγ a protein with tubulin of PIP2 in M.M. 1994; PubMed Scopus Google PIP2 was a of for min in on and with tubulin for min at in a and the was with at for the membrane with and in with PubMed Scopus Google of the was and the or to by or of Tubulin in SK-N-SH cells with or and with PBS, and of cells in on was and the for the at with on and (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google was at at at at and the with of the and cytosolic was by and in a Gβγ and Gβ1γ2 and in a or or of well from SK-N-SH neuroblastoma for Purified Gβγ and the at a protein of SK-N-SH with in for at with at for min at and the membrane membrane with the tubulin in for min at and was with specific or at with that dimeric tubulin a is polymerization at In addition, tubulin or Gβγ, specific complexes with and was to by and of the protein the of (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar) and was of and J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar) was in and Gβ1γ2 a protein of in for min at was to the at the of that and the was for 2 at J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google was by the of by by and of the protein and used for of membrane protein was with a J.S. Rasenick M.M. J. Biol. 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Wilson L. Rasenick M.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google we that interaction of tubulin with Gα regulates signaling S. Rasenick M.M. Biochemistry. 1994; 33: 9800-9805Crossref PubMed Scopus (71) Google Scholar, 5Wang N. Yan K. Rasenick M.M. J. Biol. Chem. 1990; 265: 1239-1242Abstract Full Text PDF PubMed Google Scholar, Cell Biol. PubMed Scopus Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1994; Full Text PDF PubMed Google the of Gβγ in signaling Gβγ to the translocation of signaling to signaling A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. J. M.M. R.J. PubMed Scopus Google Scholar, A. S. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google we to Gβγ also the recruitment of tubulin to the plasma membrane in response to carbachol stimulation (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google from SK-N-SH neuroblastoma cells with in the or of carbachol Gβγ J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google association of tubulin with SK-N-SH increased by after carbachol stimulation. Gβγ complexes in the the carbachol-induced in tubulin association with the membrane was the Gβγ on the association of tubulin with the membrane. Binding of Gβγ and PIP2 to Tubulin Tubulin with the PIP2 J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google to a membrane J.S. J. Rasenick M.M. J. Neurosci. PubMed Google binding of PIP2 and Gβγ to the of membrane association in response to agonist stimulation K. R.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google PIP2 and Gβγ from cells muscarinic Gαq and PLCβ1, with in the or of Gβγ, PIP2 Gαq is in this we Gβγ with the of the Gβγ to the stimulation of muscarinic receptors by carbachol tubulin to with the membrane and Gαq in Gβγ inhibited this by and Gαq transactivation was in the of PIP2 and PIP2 also the of Gβγ that with the membrane K. R.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google complexes to the membrane in response to agonist stimulation. Gβ 2 of Tubulin by Tubulin the carbachol-induced association of tubulin with the plasma membrane. carbachol stimulation also to microtubule (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google a for Gβγ inhibition of is microtubule S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google tubulin was with Gβ1γ2 with SK-N-SH that microtubule polymerization increased in the of Gβ1γ2 of tubulin was in microtubules or tubulin of tubulin with the SK-N-SH might to membrane association of tubulin of Gβγ with the of carbachol stimulation. stimulation the of membrane-associated tubulin whereas the microtubule microtubule membrane association of was observed in the of carbachol and Gβ1γ2 subunits. Gβ1γ2 to tubulin polymerization and from association with the plasma membrane in response to agonist stimulation. is with the of Gβγ with S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google It is also with the that microtubule polymerization PLCβ1 signaling Gβγ the of by of Tubulin of tubulin PLCβ1 through association of tubulin with at the cell membrane J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. J. Rasenick M.M. J. Neurosci. PubMed Google of PLCβ1 to PIP2, of Gαq coupling to PLCβ1, to for this inhibition J.S. J. Rasenick M.M. J. Neurosci. PubMed Google to Gβγ binding of tubulin dimers also this was used in Gαq activate PLCβ1 (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google PLCβ1 activation was in the and of GTPγS, tubulin-GDP, and Gβ1γ2 in muscarinic and PLCβ1 Both carbachol and increased increased the activation of PLCβ1 the that of Gαq and PLCβ1 in the cells increased the PLCβ1 the agonist stimulation of PLCβ1, muscarinic receptors of membrane the or Gβ1γ2 the of Gβ1γ2 also carbachol activation of PLCβ1 of the of suggested that Gβ1γ2 membrane PLCβ1 signaling. PLCβ1 was inhibited by by that tubulin with Gαq (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google was with Gβ1γ2 this inhibition was by the binding of tubulin to Gβ1γ2 to membrane association and in the regulation of for control of PLCβ1 inhibition by high tubulin was suggested by Gβ 2 with Gαq with, and is transactivated (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Gβγ microtubules S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google of promote microtubule Gβγ interact with tubulin at to Gβ1γ2 preferentially to Purified Gβ1γ2 was with tubulin-GDP, or tubulin of coimmunoprecipitation was with both and In both coimmunoprecipitation of Gβ1γ2 with was of Gβ1γ2 with was that that with tubulin-GTP. to the coimmunoprecipitation of with was that of tubulin-GTP. from SK-N-SH neuroblastoma with or coimmunoprecipitation of Gβ1γ2 with was coimmunoprecipitation was that of Gαq interacted with, and was Gβ1γ2 interacted preferentially with the of tubulin. It also that Gβ1γ2 in the γ2 polypeptide is promote microtubule S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google this is for Gβγ interaction with is for microtubule coimmunoprecipitation of with and is to a J. Biol. Chem. Full Text PDF PubMed Google or is that with the tubulin suggested that of the subunit is for interaction with the of dimeric tubulin. of Gβγ and interaction Gβ1γ2 and was by was with Gβ1γ2 or by of the with the EDC, and with or with the of Gβ1γ2 dimers of and a with that of complexes of with the with the revealed the of both tubulin and tubulin dimers at was also the interaction tubulin and the J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar) complexes by at was and interaction with G protein subunits. It also that the of this interaction was of in SK-N-SH tubulin and Gβγ interact with to at specific In addition, interaction is spatiotemporal colocalization in the SK-N-SH cells by confocal microscopy and after carbachol stimulation Tubulin is to PLCβ1 signaling in this neuroblastoma cell (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google In SK-N-SH tubulin and Gβ colocalized in the was at the membrane of the resting cells. after carbachol microtubules and tubulin translocated to the cell membrane 2 Gβ and tubulin colocalized at the membrane. Fifteen to carbachol exposure, tubulin and Gβ colocalized in vesicle-like structures in the association of tubulin with the plasma membrane was at this (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. J. Rasenick M.M. J. Neurosci. PubMed Google carbachol-induced of tubulin was inhibited by by coimmunoprecipitation studies of SK-N-SH cells after carbachol stimulation Gβ/tubulin complexes from SK-N-SH by with control Fifteen complexes observed in the by with control both confocal microscopy and coimmunoprecipitation in cells interaction tubulin and Gβ at the plasma membrane well translocation of complexes to the cell at the offset of signaling. of Tubulin and with used that or tubulin to tubulin translocation and colocalization with Gβ in response to agonist stimulation. observed J.S. J. Rasenick M.M. J. Neurosci. PubMed Google SK-N-SH cell microtubules and increased the of membrane-associated tubulin by well with of tubulin the plasma membrane of carbachol stimulation and colocalization in the was also the the microtubule and membrane-associated tubulin by from microtubules and tubulin colocalization with membrane Gβ and In addition, the observed after 2 min of carbachol also tubulin. the that tubulin dimers translocated to the plasma membrane in response to carbachol stimulation might through Gβγ and and are by G protein βγ Annu. Rev. 1997; PubMed Scopus Google and Rev. PubMed Scopus Google membrane association of and Gβγ M.M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, S. S. A. PubMed Scopus Google Gβγ also for receptors to the Gα binding and the GDP S. S. A. PubMed Scopus Google Gβγ is in the of cytosolic well PLCβ1, to the membrane of the cell A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. J. M.M. R.J. PubMed Scopus Google Scholar, A. S. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Gβγ for this M.M. J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Scholar, M.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google to microtubules and promote microtubule S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar) Gβγ the of the the and the of the in of J. Cell Biol. PubMed Scopus Google in the Gβγ interaction with tubulin is and for both of tubulin in PLCβ1 signaling and of the microtubule signaling and cytoskeletal is of Gβ1γ2 to tubulin regulation of PLCβ1 to interaction with Both coimmunoprecipitation and the of this interaction and was in to the Gα and interacted preferentially with (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar, 4Roychowdhury S. Rasenick M.M. Biochemistry. 1994; 33: 9800-9805Crossref PubMed Scopus (71) Google Scholar, 5Wang N. Yan K. Rasenick M.M. J. Biol. Chem. 1990; 265: 1239-1242Abstract Full Text PDF PubMed Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. Rasenick M.M. J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar, J.S. J. Rasenick M.M. J. Neurosci. PubMed Google Scholar, S. N. Rasenick M.M. Biochemistry. PubMed Scopus Google Scholar, K. Rasenick M.M. J. PubMed Scopus Google a by receptor activation membrane association of to Gαq transactivation and of PLCβ1 signaling. is by internalization of complexes at the offset of signaling. also the microtubule by S. Wilson L. Rasenick M.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar) and the increased microtubule polymerization by Gβ1γ2 S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar) also that the of binding was with Gβ J. Biol. Chem. 1994; Full Text PDF PubMed Google to both microtubules and tubulin dimers L. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google of dimeric binding is of tubulin in PLCβ1 signaling by Gβγ was by the Gβγ membrane association of tubulin and Gαq transactivation by both in the and of PIP2 2 and membrane association of tubulin and microtubule during the of carbachol Gβγ the membrane binding of tubulin microtubule polymerization In addition, Gβγ binding of the inhibition of PLCβ1 by this tubulin (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google Scholar) the of Gβγ in the of tubulin. also suggested in the cytosolic Gβγ in tubulin by microtubule in response to PubMed Scopus Google observed that in of by binding of that tubulin to the membrane and inhibited PLCβ1 J.S. J. Rasenick M.M. J. Neurosci. PubMed Google Gβγ might the inhibition of PLCβ1 by increased of J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J.S. J. Rasenick M.M. J. Neurosci. PubMed Google to colchicine, microtubule and the microtubule also inhibited PLCβ1 signaling was dimeric transactivated Gαq (3Popova J.S. Rasenick M.M. J. Neurosci. 2000; 20: 2774-2782Crossref PubMed Google in to microtubule was for the inhibition of PLCβ1 observed after pretreatment of the cells also the of tubulin regulation of PLCβ1 signaling. In SK-N-SH tubulin colocalization with Gβ at the membrane and in the after carbachol microtubule tubulin translocation to the plasma membrane, and colocalization of tubulin with membrane Gβ observed and Fifteen after carbachol tubulin and Gβ colocalized in vesicle-like structures in the by coimmunoprecipitation studies SK-N-SH cells and after carbachol Gβγ/tubulin complexes from cell membrane at 2 min of carbachol at min from the cell cytosol. SK-N-SH cells with the and by confocal tubulin translocation to the plasma membrane or colocalization with membrane Gβ was observed after agonist stimulation and tubulin was in the of the cells with the microtubule-depolymerizing increased this was to carbachol cytosolic and we that PLCβ1, Gαq J.S. J. Rasenick M.M. J. Neurosci. PubMed Google that that tubulin is with from cells in in cells by confocal is tubulin association with to cell J. PubMed Scopus Google also the that tubulin dimers to the membrane to signaling in response to agonist stimulation. studies and confocal microscopy of SK-N-SH well with and colchicine, the Gβγ tubulin translocation to the plasma membrane, might tubulin this by that a is in this S. and in of cells tubulin and Gβγ J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. Cell Biol. PubMed Scopus Google Scholar, S. J. PubMed Scopus Google and of a in Cell Biol. Scholar, PubMed Scopus Google In addition, Gβγ to the of muscarinic receptors J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google to through a A. S. J. Biol. Chem. Full Text PDF PubMed Google Scholar, J. PubMed Scopus Google Scholar, J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google tubulin to the in response to agonist stimulation J. PubMed Scopus Google to the muscarinic receptor J.S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google tubulin might with muscarinic receptors through Gβγ or Gβγ suggested to protein the of G protein-coupled receptors with signaling J. Biol. Chem. 2000; Full Text Full Text PDF PubMed Scopus Google Gβγ observed in the N. 2000; PubMed Scopus Google Scholar) and is with microtubules from S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google in this that Gβγ membrane translocation of tubulin in response to agonist stimulation of Gβγ in this Gβγ to the translocation of and J. J. M.M. R.J. PubMed Scopus Google Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google is for recruitment to the membrane of the cell A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google GRK2, and Gβγ to S. Rasenick M.M. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google might a signaling that to the membrane in response to agonist stimulation. It that both signal and sequestration of signaling translocation of of signal are protein R.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Gα and Gβγ in is to Gα observed to with microtubules in from cells T.J. Rasenick M.M. J. PubMed Scopus Google Scholar) and a to microtubules transactivation of Gα by cells from Rasenick M.M. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google It is in this that microtubules that are a interaction Gα and Gβγ in microtubules in a to microtubules in a 1994; Scopus Google In we that Gβγ tubulin to the plasma membrane for the regulation of PLCβ1 signaling promote microtubule Gβ1γ2 preferentially with tubulin-GDP, this for of the cell microtubules in response to in In inhibition of PLCβ1 by high tubulin by tubulin interaction with Gβγ and of that tubulin microtubules. during the of agonist membrane Gβγ sequestration of membrane-associated tubulin and this a for signaling and the of the microtubule of to the regulation of and cell and in cell A. J. S. J. and for the of S. A. K. K. and J. are for for