Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits

Pamela M. Taylor, Christopher N. Connolly, Josef T. Kittler, George H. Gorrie, Alistair Hosie, Trevor G. Smart, Stephen J. Moss

    Research output: Contribution to journalArticle

    63 Citations (Scopus)

    Abstract

    GABA(A) receptors can be constructed from a range of differing subunit isoforms: alpha, beta, gamma, delta, and epsilon. Expression studies have revealed that production of GABA-gated channels is achieved after coexpression of alpha and beta subunits. The expression of a gamma subunit isoform is essential to confer benzodiazepine sensitivity on the expressed receptor. However, how the specificity of subunit interactions is controlled during receptor assembly remains unknown. Here we demonstrate that residues 58-67 within alpha subunit isoforms are important in the assembly of receptors comprised of alpha beta and alpha beta gamma subunits. Deletion of these residues from the alpha 1 or alpha 6 subunits results in retention of either alpha subunit isoform in the endoplasmic reticulum on coexpression with the beta 3, or beta 3 and gamma 2 subunits. Immunoprecipitation revealed that residues 58-67 mediated oligomerization of the alpha 1 and alpha 3 subunits, but were without affect on the production of alpha/gamma complexes. Within this domain, glutamine 67 was of central importance in mediating the production of functional alpha 1 beta 3 receptors. Mutation of this residue resulted in a drastic decrease in the cell surface expression of alpha 1 beta 3 receptors and the resulting expression of beta 3 homomers. Sucrose density gradient centrifugation revealed that this residue was important for the production of a 9S alpha 1 beta 3 complex representing functional GABA(A) receptors.

    Therefore, our studies detail residues that specify GABA(A) receptor alpha beta subunit interactions. This domain, which is conserved in all alpha subunit isoforms, will therefore play a critical role in the assembly of GABA(A) receptors composed of alpha beta and alpha beta gamma subunits.

    Original languageEnglish
    Pages (from-to)1297-1306
    Number of pages10
    JournalJournal of Neuroscience
    Volume20
    Issue number4
    Publication statusPublished - 2000

    Cite this

    Taylor, P. M., Connolly, C. N., Kittler, J. T., Gorrie, G. H., Hosie, A., Smart, T. G., & Moss, S. J. (2000). Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits. Journal of Neuroscience, 20(4), 1297-1306.
    Taylor, Pamela M. ; Connolly, Christopher N. ; Kittler, Josef T. ; Gorrie, George H. ; Hosie, Alistair ; Smart, Trevor G. ; Moss, Stephen J. / Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits. In: Journal of Neuroscience. 2000 ; Vol. 20, No. 4. pp. 1297-1306.
    @article{2420430f70824985a61dc3ece691130b,
    title = "Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits",
    abstract = "GABA(A) receptors can be constructed from a range of differing subunit isoforms: alpha, beta, gamma, delta, and epsilon. Expression studies have revealed that production of GABA-gated channels is achieved after coexpression of alpha and beta subunits. The expression of a gamma subunit isoform is essential to confer benzodiazepine sensitivity on the expressed receptor. However, how the specificity of subunit interactions is controlled during receptor assembly remains unknown. Here we demonstrate that residues 58-67 within alpha subunit isoforms are important in the assembly of receptors comprised of alpha beta and alpha beta gamma subunits. Deletion of these residues from the alpha 1 or alpha 6 subunits results in retention of either alpha subunit isoform in the endoplasmic reticulum on coexpression with the beta 3, or beta 3 and gamma 2 subunits. Immunoprecipitation revealed that residues 58-67 mediated oligomerization of the alpha 1 and alpha 3 subunits, but were without affect on the production of alpha/gamma complexes. Within this domain, glutamine 67 was of central importance in mediating the production of functional alpha 1 beta 3 receptors. Mutation of this residue resulted in a drastic decrease in the cell surface expression of alpha 1 beta 3 receptors and the resulting expression of beta 3 homomers. Sucrose density gradient centrifugation revealed that this residue was important for the production of a 9S alpha 1 beta 3 complex representing functional GABA(A) receptors.Therefore, our studies detail residues that specify GABA(A) receptor alpha beta subunit interactions. This domain, which is conserved in all alpha subunit isoforms, will therefore play a critical role in the assembly of GABA(A) receptors composed of alpha beta and alpha beta gamma subunits.",
    author = "Taylor, {Pamela M.} and Connolly, {Christopher N.} and Kittler, {Josef T.} and Gorrie, {George H.} and Alistair Hosie and Smart, {Trevor G.} and Moss, {Stephen J.}",
    year = "2000",
    language = "English",
    volume = "20",
    pages = "1297--1306",
    journal = "Journal of Neuroscience",
    issn = "0270-6474",
    publisher = "Society for Neuroscience",
    number = "4",

    }

    Taylor, PM, Connolly, CN, Kittler, JT, Gorrie, GH, Hosie, A, Smart, TG & Moss, SJ 2000, 'Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits', Journal of Neuroscience, vol. 20, no. 4, pp. 1297-1306.

    Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits. / Taylor, Pamela M.; Connolly, Christopher N.; Kittler, Josef T.; Gorrie, George H.; Hosie, Alistair; Smart, Trevor G.; Moss, Stephen J.

    In: Journal of Neuroscience, Vol. 20, No. 4, 2000, p. 1297-1306.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Identification of residues within GABAA receptor subunits that mediate specific assembly with receptor β subunits

    AU - Taylor, Pamela M.

    AU - Connolly, Christopher N.

    AU - Kittler, Josef T.

    AU - Gorrie, George H.

    AU - Hosie, Alistair

    AU - Smart, Trevor G.

    AU - Moss, Stephen J.

    PY - 2000

    Y1 - 2000

    N2 - GABA(A) receptors can be constructed from a range of differing subunit isoforms: alpha, beta, gamma, delta, and epsilon. Expression studies have revealed that production of GABA-gated channels is achieved after coexpression of alpha and beta subunits. The expression of a gamma subunit isoform is essential to confer benzodiazepine sensitivity on the expressed receptor. However, how the specificity of subunit interactions is controlled during receptor assembly remains unknown. Here we demonstrate that residues 58-67 within alpha subunit isoforms are important in the assembly of receptors comprised of alpha beta and alpha beta gamma subunits. Deletion of these residues from the alpha 1 or alpha 6 subunits results in retention of either alpha subunit isoform in the endoplasmic reticulum on coexpression with the beta 3, or beta 3 and gamma 2 subunits. Immunoprecipitation revealed that residues 58-67 mediated oligomerization of the alpha 1 and alpha 3 subunits, but were without affect on the production of alpha/gamma complexes. Within this domain, glutamine 67 was of central importance in mediating the production of functional alpha 1 beta 3 receptors. Mutation of this residue resulted in a drastic decrease in the cell surface expression of alpha 1 beta 3 receptors and the resulting expression of beta 3 homomers. Sucrose density gradient centrifugation revealed that this residue was important for the production of a 9S alpha 1 beta 3 complex representing functional GABA(A) receptors.Therefore, our studies detail residues that specify GABA(A) receptor alpha beta subunit interactions. This domain, which is conserved in all alpha subunit isoforms, will therefore play a critical role in the assembly of GABA(A) receptors composed of alpha beta and alpha beta gamma subunits.

    AB - GABA(A) receptors can be constructed from a range of differing subunit isoforms: alpha, beta, gamma, delta, and epsilon. Expression studies have revealed that production of GABA-gated channels is achieved after coexpression of alpha and beta subunits. The expression of a gamma subunit isoform is essential to confer benzodiazepine sensitivity on the expressed receptor. However, how the specificity of subunit interactions is controlled during receptor assembly remains unknown. Here we demonstrate that residues 58-67 within alpha subunit isoforms are important in the assembly of receptors comprised of alpha beta and alpha beta gamma subunits. Deletion of these residues from the alpha 1 or alpha 6 subunits results in retention of either alpha subunit isoform in the endoplasmic reticulum on coexpression with the beta 3, or beta 3 and gamma 2 subunits. Immunoprecipitation revealed that residues 58-67 mediated oligomerization of the alpha 1 and alpha 3 subunits, but were without affect on the production of alpha/gamma complexes. Within this domain, glutamine 67 was of central importance in mediating the production of functional alpha 1 beta 3 receptors. Mutation of this residue resulted in a drastic decrease in the cell surface expression of alpha 1 beta 3 receptors and the resulting expression of beta 3 homomers. Sucrose density gradient centrifugation revealed that this residue was important for the production of a 9S alpha 1 beta 3 complex representing functional GABA(A) receptors.Therefore, our studies detail residues that specify GABA(A) receptor alpha beta subunit interactions. This domain, which is conserved in all alpha subunit isoforms, will therefore play a critical role in the assembly of GABA(A) receptors composed of alpha beta and alpha beta gamma subunits.

    M3 - Article

    VL - 20

    SP - 1297

    EP - 1306

    JO - Journal of Neuroscience

    JF - Journal of Neuroscience

    SN - 0270-6474

    IS - 4

    ER -