Abstract
Modulation of A-type voltage-gated K+ channels can produce plastic changes in neuronal signaling. It was shown that the delayed-rectifier Kv1.1 channel can be converted to A-type upon association with Kvbeta1.1 subunits; the conversion is only partial and is modulated by phosphorylation and microfilaments. Here we show that, in Xenopus oocytes, expression of Gbeta1gamma2 subunits concomitantly with the channel (composed of Kv1.1 and Kvbeta1.1 subunits), but not after the channel's expression in the plasma membrane, increases the extent of conversion to A-type. Conversely, scavenging endogenous Gbetagamma by co-expression of the C-terminal fragment of the beta-adrenergic receptor kinase reduces the extent of conversion to A-type. The effect of Gbetagamma co-expression is occluded by treatment with dihydrocytochalasin B, a microfilament-disrupting agent shown previously by us to enhance the extent of conversion to A-type, and by overexpression of Kvbeta1.1. Gbeta1gamma2 subunits interact directly with GST fusion fragments of Kv1.1 and Kvbeta1.1. Co-expression of Gbeta1gamma2 causes co-immunoprecipitation with Kv1.1 of more Kvbeta1.1 subunits. Thus, we suggest that Gbeta1gamma2 directly affects the interaction between Kv1.1 and Kvbeta1.1 during channel assembly which, in turn, disrupts the ability of the channel to interact with microfilaments, resulting in an increased extent of A-type conversion.
Full Text
The Full Text of this article is available as a PDF (631.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Armstrong C. M., Bezanilla F. Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol. 1977 Nov;70(5):567–590. doi: 10.1085/jgp.70.5.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bomsel M., Mostov K. Role of heterotrimeric G proteins in membrane traffic. Mol Biol Cell. 1992 Dec;3(12):1317–1328. doi: 10.1091/mbc.3.12.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carlson K. E., Woolkalis M. J., Newhouse M. G., Manning D. R. Fractionation of the beta subunit common to guanine nucleotide-binding regulatory proteins with the cytoskeleton. Mol Pharmacol. 1986 Nov;30(5):463–468. [PubMed] [Google Scholar]
- Chen J., DeVivo M., Dingus J., Harry A., Li J., Sui J., Carty D. J., Blank J. L., Exton J. H., Stoffel R. H. A region of adenylyl cyclase 2 critical for regulation by G protein beta gamma subunits. Science. 1995 May 26;268(5214):1166–1169. doi: 10.1126/science.7761832. [DOI] [PubMed] [Google Scholar]
- Clapham D. E., Neer E. J. G protein beta gamma subunits. Annu Rev Pharmacol Toxicol. 1997;37:167–203. doi: 10.1146/annurev.pharmtox.37.1.167. [DOI] [PubMed] [Google Scholar]
- Connor J. A., Stevens C. F. Voltage clamp studies of a transient outward membrane current in gastropod neural somata. J Physiol. 1971 Feb;213(1):21–30. doi: 10.1113/jphysiol.1971.sp009365. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crow T. Cellular and molecular analysis of associative learning and memory in Hermissenda. Trends Neurosci. 1988 Apr;11(4):136–147. doi: 10.1016/0166-2236(88)90138-5. [DOI] [PubMed] [Google Scholar]
- Dascal N., Doupnik C. A., Ivanina T., Bausch S., Wang W., Lin C., Garvey J., Chavkin C., Lester H. A., Davidson N. Inhibition of function in Xenopus oocytes of the inwardly rectifying G-protein-activated atrial K channel (GIRK1) by overexpression of a membrane-attached form of the C-terminal tail. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6758–6762. doi: 10.1073/pnas.92.15.6758. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dascal N. Signalling via the G protein-activated K+ channels. Cell Signal. 1997 Dec;9(8):551–573. doi: 10.1016/s0898-6568(97)00095-8. [DOI] [PubMed] [Google Scholar]
- Dolphin A. C. Mechanisms of modulation of voltage-dependent calcium channels by G proteins. J Physiol. 1998 Jan 1;506(Pt 1):3–11. doi: 10.1111/j.1469-7793.1998.003bx.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Heinemann S., Rettig J., Scott V., Parcej D. N., Lorra C., Dolly J., Pongs O. The inactivation behaviour of voltage-gated K-channels may be determined by association of alpha- and beta-subunits. J Physiol Paris. 1994;88(3):173–180. doi: 10.1016/0928-4257(94)90003-5. [DOI] [PubMed] [Google Scholar]
- Hoshi T., Zagotta W. N., Aldrich R. W. Biophysical and molecular mechanisms of Shaker potassium channel inactivation. Science. 1990 Oct 26;250(4980):533–538. doi: 10.1126/science.2122519. [DOI] [PubMed] [Google Scholar]
- Inglese J., Luttrell L. M., Iñiguez-Lluhi J. A., Touhara K., Koch W. J., Lefkowitz R. J. Functionally active targeting domain of the beta-adrenergic receptor kinase: an inhibitor of G beta gamma-mediated stimulation of type II adenylyl cyclase. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3637–3641. doi: 10.1073/pnas.91.9.3637. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ivanina T., Perets T., Thornhill W. B., Levin G., Dascal N., Lotan I. Phosphorylation by protein kinase A of RCK1 K+ channels expressed in Xenopus oocytes. Biochemistry. 1994 Jul 26;33(29):8786–8792. doi: 10.1021/bi00195a021. [DOI] [PubMed] [Google Scholar]
- Jamora C., Takizawa P. A., Zaarour R. F., Denesvre C., Faulkner D. J., Malhotra V. Regulation of Golgi structure through heterotrimeric G proteins. Cell. 1997 Nov 28;91(5):617–626. doi: 10.1016/s0092-8674(00)80449-3. [DOI] [PubMed] [Google Scholar]
- Jing J., Peretz T., Singer-Lahat D., Chikvashvili D., Thornhill W. B., Lotan I. Inactivation of a voltage-dependent K+ channel by beta subunit. Modulation by a phosphorylation-dependent interaction between the distal C terminus of alpha subunit and cytoskeleton. J Biol Chem. 1997 May 30;272(22):14021–14024. doi: 10.1074/jbc.272.22.14021. [DOI] [PubMed] [Google Scholar]
- Kim E., Niethammer M., Rothschild A., Jan Y. N., Sheng M. Clustering of Shaker-type K+ channels by interaction with a family of membrane-associated guanylate kinases. Nature. 1995 Nov 2;378(6552):85–88. doi: 10.1038/378085a0. [DOI] [PubMed] [Google Scholar]
- Koch W. J., Hawes B. E., Inglese J., Luttrell L. M., Lefkowitz R. J. Cellular expression of the carboxyl terminus of a G protein-coupled receptor kinase attenuates G beta gamma-mediated signaling. J Biol Chem. 1994 Feb 25;269(8):6193–6197. [PubMed] [Google Scholar]
- Koch W. J., Inglese J., Stone W. C., Lefkowitz R. J. The binding site for the beta gamma subunits of heterotrimeric G proteins on the beta-adrenergic receptor kinase. J Biol Chem. 1993 Apr 15;268(11):8256–8260. [PubMed] [Google Scholar]
- Levin G., Chikvashvili D., Singer-Lahat D., Peretz T., Thornhill W. B., Lotan I. Phosphorylation of a K+ channel alpha subunit modulates the inactivation conferred by a beta subunit. Involvement of cytoskeleton. J Biol Chem. 1996 Nov 15;271(46):29321–29328. doi: 10.1074/jbc.271.46.29321. [DOI] [PubMed] [Google Scholar]
- Levin G., Keren T., Peretz T., Chikvashvili D., Thornhill W. B., Lotan I. Regulation of RCK1 currents with a cAMP analog via enhanced protein synthesis and direct channel phosphorylation. J Biol Chem. 1995 Jun 16;270(24):14611–14618. doi: 10.1074/jbc.270.24.14611. [DOI] [PubMed] [Google Scholar]
- Levy M., Jing J., Chikvashvili D., Thornhill W. B., Lotan I. Activation of a metabotropic glutamate receptor and protein kinase C reduce the extent of inactivation of the K+ channel Kv1.1/Kvbeta1.1 via dephosphorylation of Kv1.1. J Biol Chem. 1998 Mar 13;273(11):6495–6502. doi: 10.1074/jbc.273.11.6495. [DOI] [PubMed] [Google Scholar]
- Li M., Jan Y. N., Jan L. Y. Specification of subunit assembly by the hydrophilic amino-terminal domain of the Shaker potassium channel. Science. 1992 Aug 28;257(5074):1225–1230. doi: 10.1126/science.1519059. [DOI] [PubMed] [Google Scholar]
- Lin C. T., Wu H. C., Cheng H. F., Chang J. T., Chang K. J. Identification of beta-subunit of GTP-binding regulatory protein in mitotic spindle. Lab Invest. 1992 Dec;67(6):770–778. [PubMed] [Google Scholar]
- Ma J. Y., Catterall W. A., Scheuer T. Persistent sodium currents through brain sodium channels induced by G protein betagamma subunits. Neuron. 1997 Aug;19(2):443–452. doi: 10.1016/s0896-6273(00)80952-6. [DOI] [PubMed] [Google Scholar]
- MacKinnon R., Aldrich R. W., Lee A. W. Functional stoichiometry of Shaker potassium channel inactivation. Science. 1993 Oct 29;262(5134):757–759. doi: 10.1126/science.7694359. [DOI] [PubMed] [Google Scholar]
- Muller L., Picart R., Barret A., Bockaert J., Homburger V., Tougard C. Identification of multiple subunits of heterotrimeric G proteins on the membrane of secretory granules in rat prolactin anterior pituitary cells. Mol Cell Neurosci. 1994 Dec;5(6):556–566. doi: 10.1006/mcne.1994.1068. [DOI] [PubMed] [Google Scholar]
- Nagaya N., Papazian D. M. Potassium channel alpha and beta subunits assemble in the endoplasmic reticulum. J Biol Chem. 1997 Jan 31;272(5):3022–3027. doi: 10.1074/jbc.272.5.3022. [DOI] [PubMed] [Google Scholar]
- Neubig R. R. Membrane organization in G-protein mechanisms. FASEB J. 1994 Sep;8(12):939–946. doi: 10.1096/fasebj.8.12.8088459. [DOI] [PubMed] [Google Scholar]
- Rettig J., Heinemann S. H., Wunder F., Lorra C., Parcej D. N., Dolly J. O., Pongs O. Inactivation properties of voltage-gated K+ channels altered by presence of beta-subunit. Nature. 1994 May 26;369(6478):289–294. doi: 10.1038/369289a0. [DOI] [PubMed] [Google Scholar]
- Reuveny E., Slesinger P. A., Inglese J., Morales J. M., Iñiguez-Lluhi J. A., Lefkowitz R. J., Bourne H. R., Jan Y. N., Jan L. Y. Activation of the cloned muscarinic potassium channel by G protein beta gamma subunits. Nature. 1994 Jul 14;370(6485):143–146. doi: 10.1038/370143a0. [DOI] [PubMed] [Google Scholar]
- Rhodes K. J., Keilbaugh S. A., Barrezueta N. X., Lopez K. L., Trimmer J. S. Association and colocalization of K+ channel alpha- and beta-subunit polypeptides in rat brain. J Neurosci. 1995 Jul;15(7 Pt 2):5360–5371. doi: 10.1523/JNEUROSCI.15-07-05360.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rhodes K. J., Monaghan M. M., Barrezueta N. X., Nawoschik S., Bekele-Arcuri Z., Matos M. F., Nakahira K., Schechter L. E., Trimmer J. S. Voltage-gated K+ channel beta subunits: expression and distribution of Kv beta 1 and Kv beta 2 in adult rat brain. J Neurosci. 1996 Aug 15;16(16):4846–4860. doi: 10.1523/JNEUROSCI.16-16-04846.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rudy B. Diversity and ubiquity of K channels. Neuroscience. 1988 Jun;25(3):729–749. doi: 10.1016/0306-4522(88)90033-4. [DOI] [PubMed] [Google Scholar]
- Scannevin R. H., Trimmer J. S. Cytoplasmic domains of voltage-sensitive K+ channels involved in mediating protein-protein interactions. Biochem Biophys Res Commun. 1997 Mar 27;232(3):585–589. doi: 10.1006/bbrc.1997.6333. [DOI] [PubMed] [Google Scholar]
- Sewing S., Roeper J., Pongs O. Kv beta 1 subunit binding specific for shaker-related potassium channel alpha subunits. Neuron. 1996 Feb;16(2):455–463. doi: 10.1016/s0896-6273(00)80063-x. [DOI] [PubMed] [Google Scholar]
- Shen N. V., Chen X., Boyer M. M., Pfaffinger P. J. Deletion analysis of K+ channel assembly. Neuron. 1993 Jul;11(1):67–76. doi: 10.1016/0896-6273(93)90271-r. [DOI] [PubMed] [Google Scholar]
- Shen N. V., Pfaffinger P. J. Molecular recognition and assembly sequences involved in the subfamily-specific assembly of voltage-gated K+ channel subunit proteins. Neuron. 1995 Mar;14(3):625–633. doi: 10.1016/0896-6273(95)90319-4. [DOI] [PubMed] [Google Scholar]
- Sheng M., Liao Y. J., Jan Y. N., Jan L. Y. Presynaptic A-current based on heteromultimeric K+ channels detected in vivo. Nature. 1993 Sep 2;365(6441):72–75. doi: 10.1038/365072a0. [DOI] [PubMed] [Google Scholar]
- Shi G., Nakahira K., Hammond S., Rhodes K. J., Schechter L. E., Trimmer J. S. Beta subunits promote K+ channel surface expression through effects early in biosynthesis. Neuron. 1996 Apr;16(4):843–852. doi: 10.1016/s0896-6273(00)80104-x. [DOI] [PubMed] [Google Scholar]
- Stow J. L., de Almeida J. B., Narula N., Holtzman E. J., Ercolani L., Ausiello D. A. A heterotrimeric G protein, G alpha i-3, on Golgi membranes regulates the secretion of a heparan sulfate proteoglycan in LLC-PK1 epithelial cells. J Cell Biol. 1991 Sep;114(6):1113–1124. doi: 10.1083/jcb.114.6.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stühmer W., Ruppersberg J. P., Schröter K. H., Sakmann B., Stocker M., Giese K. P., Perschke A., Baumann A., Pongs O. Molecular basis of functional diversity of voltage-gated potassium channels in mammalian brain. EMBO J. 1989 Nov;8(11):3235–3244. doi: 10.1002/j.1460-2075.1989.tb08483.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Veh R. W., Lichtinghagen R., Sewing S., Wunder F., Grumbach I. M., Pongs O. Immunohistochemical localization of five members of the Kv1 channel subunits: contrasting subcellular locations and neuron-specific co-localizations in rat brain. Eur J Neurosci. 1995 Nov 1;7(11):2189–2205. doi: 10.1111/j.1460-9568.1995.tb00641.x. [DOI] [PubMed] [Google Scholar]
- Wang H., Kunkel D. D., Martin T. M., Schwartzkroin P. A., Tempel B. L. Heteromultimeric K+ channels in terminal and juxtaparanodal regions of neurons. Nature. 1993 Sep 2;365(6441):75–79. doi: 10.1038/365075a0. [DOI] [PubMed] [Google Scholar]
- Wang H., Kunkel D. D., Schwartzkroin P. A., Tempel B. L. Localization of Kv1.1 and Kv1.2, two K channel proteins, to synaptic terminals, somata, and dendrites in the mouse brain. J Neurosci. 1994 Aug;14(8):4588–4599. doi: 10.1523/JNEUROSCI.14-08-04588.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickman K., Clapham D. E. Ion channel regulation by G proteins. Physiol Rev. 1995 Oct;75(4):865–885. doi: 10.1152/physrev.1995.75.4.865. [DOI] [PubMed] [Google Scholar]
- Xu J., Li M. Kvbeta2 inhibits the Kvbeta1-mediated inactivation of K+ channels in transfected mammalian cells. J Biol Chem. 1997 May 2;272(18):11728–11735. doi: 10.1074/jbc.272.18.11728. [DOI] [PubMed] [Google Scholar]
- Xu J., Yu W., Wright J. M., Raab R. W., Li M. Distinct functional stoichiometry of potassium channel beta subunits. Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1846–1851. doi: 10.1073/pnas.95.4.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yu W., Xu J., Li M. NAB domain is essential for the subunit assembly of both alpha-alpha and alpha-beta complexes of shaker-like potassium channels. Neuron. 1996 Feb;16(2):441–453. doi: 10.1016/s0896-6273(00)80062-8. [DOI] [PubMed] [Google Scholar]
- Zagotta W. N., Hoshi T., Aldrich R. W. Restoration of inactivation in mutants of Shaker potassium channels by a peptide derived from ShB. Science. 1990 Oct 26;250(4980):568–571. doi: 10.1126/science.2122520. [DOI] [PubMed] [Google Scholar]