Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The EMBO Journal logoLink to The EMBO Journal
. 1989 Jun;8(6):1665–1670. doi: 10.1002/j.1460-2075.1989.tb03557.x

GABAA receptor beta subunit heterogeneity: functional expression of cloned cDNAs.

S Ymer 1, P R Schofield 1, A Draguhn 1, P Werner 1, M Köhler 1, P H Seeburg 1
PMCID: PMC401007  PMID: 2548852

Abstract

Cloned cDNAs encoding two new beta subunits of the rat and bovine GABAA receptor have been isolated using a degenerate oligonucleotide probe based on a highly conserved peptide sequence in the second transmembrane domain of GABAA receptor subunits. The beta 2 and beta 3 subunits share approximately 72% sequence identity with the previously characterized beta 1 polypeptide. Northern analysis showed that both beta 2 and beta 3 mRNAs are more abundant in the brain than beta 1 mRNA. All three beta subunit encoding cDNAs were also identified in a library constructed from adrenal medulla RNA. Each beta subunit, when co-expressed in Xenopus oocytes with an alpha subunit, forms functional GABAA receptors. These results, together with the known alpha subunit heterogeneity, suggest that a variety of related but functionally distinct GABAA receptor subtypes are generated by different subunit combinations.

Full text

PDF
1665

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ballivet M., Nef P., Couturier S., Rungger D., Bader C. R., Bertrand D., Cooper E. Electrophysiology of a chick neuronal nicotinic acetylcholine receptor expressed in Xenopus oocytes after cDNA injection. Neuron. 1988 Nov;1(9):847–852. doi: 10.1016/0896-6273(88)90132-8. [DOI] [PubMed] [Google Scholar]
  2. Bormann J., Clapham D. E. gamma-Aminobutyric acid receptor channels in adrenal chromaffin cells: a patch-clamp study. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2168–2172. doi: 10.1073/pnas.82.7.2168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Braestrup C., Nielsen M. [3H]Propyl beta-carboline-3-carboxylate as a selective radioligand for the BZ1 benzodiazepine receptor subclass. J Neurochem. 1981 Aug;37(2):333–341. doi: 10.1111/j.1471-4159.1981.tb00460.x. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Criado M., Sarin V., Fox J. L., Lindstrom J. Evidence that the acetylcholine binding site is not formed by the sequence alpha 127-143 of the acetylcholine receptor. Biochemistry. 1986 May 20;25(10):2839–2846. doi: 10.1021/bi00358a015. [DOI] [PubMed] [Google Scholar]
  6. Dascal N., Landau E. M., Lass Y. Xenopus oocyte resting potential, muscarinic responses and the role of calcium and guanosine 3',5'-cyclic monophosphate. J Physiol. 1984 Jul;352:551–574. doi: 10.1113/jphysiol.1984.sp015310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Feramisco J. R., Glass D. B., Krebs E. G. Optimal spatial requirements for the location of basic residues in peptide substrates for the cyclic AMP-dependent protein kinase. J Biol Chem. 1980 May 10;255(9):4240–4245. [PubMed] [Google Scholar]
  8. Fuchs K., Möhler H., Sieghart W. Various proteins from rat brain, specifically and irreversibly labeled by [3H]flunitrazepam, are distinct alpha-subunits of the GABA-benzodiazepine receptor complex. Neurosci Lett. 1988 Aug 1;90(3):314–319. doi: 10.1016/0304-3940(88)90208-x. [DOI] [PubMed] [Google Scholar]
  9. Grandison L., Guidotti A. gamma-Aminobutyric acid receptor function in rat anterior pituitary: evidence for control of prolactin release. Endocrinology. 1979 Sep;105(3):754–759. doi: 10.1210/endo-105-3-754. [DOI] [PubMed] [Google Scholar]
  10. Grenningloh G., Gundelfinger E., Schmitt B., Betz H., Darlison M. G., Barnard E. A., Schofield P. R., Seeburg P. H. Glycine vs GABA receptors. Nature. 1987 Nov 5;330(6143):25–26. doi: 10.1038/330025b0. [DOI] [PubMed] [Google Scholar]
  11. Grenningloh G., Rienitz A., Schmitt B., Methfessel C., Zensen M., Beyreuther K., Gundelfinger E. D., Betz H. The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors. Nature. 1987 Jul 16;328(6127):215–220. doi: 10.1038/328215a0. [DOI] [PubMed] [Google Scholar]
  12. Häring P., Stähli C., Schoch P., Takács B., Staehelin T., Möhler H. Monoclonal antibodies reveal structural homogeneity of gamma-aminobutyric acid/benzodiazepine receptors in different brain areas. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4837–4841. doi: 10.1073/pnas.82.14.4837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Imoto K., Methfessel C., Sakmann B., Mishina M., Mori Y., Konno T., Fukuda K., Kurasaki M., Bujo H., Fujita Y. Location of a delta-subunit region determining ion transport through the acetylcholine receptor channel. Nature. 1986 Dec 18;324(6098):670–674. doi: 10.1038/324670a0. [DOI] [PubMed] [Google Scholar]
  14. Kataoka Y., Gutman Y., Guidotti A., Panula P., Wroblewski J., Cosenza-Murphy D., Wu J. Y., Costa E. Intrinsic GABAergic system of adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1984 May;81(10):3218–3222. doi: 10.1073/pnas.81.10.3218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leonard R. J., Labarca C. G., Charnet P., Davidson N., Lester H. A. Evidence that the M2 membrane-spanning region lines the ion channel pore of the nicotinic receptor. Science. 1988 Dec 16;242(4885):1578–1581. doi: 10.1126/science.2462281. [DOI] [PubMed] [Google Scholar]
  16. Levitan E. S., Schofield P. R., Burt D. R., Rhee L. M., Wisden W., Köhler M., Fujita N., Rodriguez H. F., Stephenson A., Darlison M. G. Structural and functional basis for GABAA receptor heterogeneity. Nature. 1988 Sep 1;335(6185):76–79. doi: 10.1038/335076a0. [DOI] [PubMed] [Google Scholar]
  17. Mamalaki C., Stephenson F. A., Barnard E. A. The GABAA/benzodiazepine receptor is a heterotetramer of homologous alpha and beta subunits. EMBO J. 1987 Mar;6(3):561–565. doi: 10.1002/j.1460-2075.1987.tb04791.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Möhler H., Battersby M. K., Richards J. G. Benzodiazepine receptor protein identified and visualized in brain tissue by a photoaffinity label. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1666–1670. doi: 10.1073/pnas.77.3.1666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pritchett D. B., Sontheimer H., Shivers B. D., Ymer S., Kettenmann H., Schofield P. R., Seeburg P. H. Importance of a novel GABAA receptor subunit for benzodiazepine pharmacology. Nature. 1989 Apr 13;338(6216):582–585. doi: 10.1038/338582a0. [DOI] [PubMed] [Google Scholar]
  20. Racagni G., Apud J. A., Locatelli V., Cocchi D., Nistico G., di Giorgio R. M., Müller E. E. GABA of CNS origin in the rat anterior pituitary inhibits prolactin secretion. Nature. 1979 Oct 18;281(5732):575–578. doi: 10.1038/281575a0. [DOI] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schofield P. R., Darlison M. G., Fujita N., Burt D. R., Stephenson F. A., Rodriguez H., Rhee L. M., Ramachandran J., Reale V., Glencorse T. A. Sequence and functional expression of the GABA A receptor shows a ligand-gated receptor super-family. Nature. 1987 Jul 16;328(6127):221–227. doi: 10.1038/328221a0. [DOI] [PubMed] [Google Scholar]
  23. Schofield P. R., Pritchett D. B., Sontheimer H., Kettenmann H., Seeburg P. H. Sequence and expression of human GABAA receptor alpha 1 and beta 1 subunits. FEBS Lett. 1989 Feb 27;244(2):361–364. doi: 10.1016/0014-5793(89)80563-0. [DOI] [PubMed] [Google Scholar]
  24. Seed B. An LFA-3 cDNA encodes a phospholipid-linked membrane protein homologous to its receptor CD2. 1987 Oct 29-Nov 4Nature. 329(6142):840–842. doi: 10.1038/329840a0. [DOI] [PubMed] [Google Scholar]
  25. Setzer D. R., McGrogan M., Nunberg J. H., Schimke R. T. Size heterogeneity in the 3' end of dihydrofolate reductase messenger RNAs in mouse cells. Cell. 1980 Nov;22(2 Pt 2):361–370. doi: 10.1016/0092-8674(80)90346-3. [DOI] [PubMed] [Google Scholar]
  26. Siegel R. E. The mRNAs encoding GABAA/benzodiazepine receptor subunits are localized in different cell populations of the bovine cerebellum. Neuron. 1988 Sep;1(7):579–584. doi: 10.1016/0896-6273(88)90107-9. [DOI] [PubMed] [Google Scholar]
  27. Sieghart W., Mayer A., Drexler G. Properties of [3H]flunitrazepam binding to different benzodiazepine binding proteins. Eur J Pharmacol. 1983 Apr 8;88(4):291–299. doi: 10.1016/0014-2999(83)90579-4. [DOI] [PubMed] [Google Scholar]
  28. Squires R. F., Benson D. I., Braestrup C., Coupet J., Klepner C. A., Myers V., Beer B. Some properties of brain specific benzodiazepine receptors: new evidence for multiple receptors. Pharmacol Biochem Behav. 1979 May;10(5):825–830. doi: 10.1016/0091-3057(79)90341-1. [DOI] [PubMed] [Google Scholar]
  29. Stephenson F. A. Understanding the GABAA receptor: a chemically gated ion channel. Biochem J. 1988 Jan 1;249(1):21–32. doi: 10.1042/bj2490021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Séquier J. M., Richards J. G., Malherbe P., Price G. W., Mathews S., Möhler H. Mapping of brain areas containing RNA homologous to cDNAs encoding the alpha and beta subunits of the rat GABAA gamma-aminobutyrate receptor. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7815–7819. doi: 10.1073/pnas.85.20.7815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tosi M., Young R. A., Hagenbüchle O., Schibler U. Multiple polyadenylation sites in a mouse alpha-amylase gene. Nucleic Acids Res. 1981 May 25;9(10):2313–2323. doi: 10.1093/nar/9.10.2313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Unnerstall J. R., Kuhar M. J., Niehoff D. L., Palacios J. M. Benzodiazepine receptors are coupled to a subpopulation of gamma-aminobutyric acid (GABA) receptors: evidence from a quantitative autoradiographic study. J Pharmacol Exp Ther. 1981 Sep;218(3):797–804. [PubMed] [Google Scholar]
  33. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  34. Voellmy R., Rungger D. Transcription of a Drosophila heat shock gene is heat-induced in Xenopus oocytes. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1776–1780. doi: 10.1073/pnas.79.6.1776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES