An ER-resident Membrane Protein Complex Regulates Nicotinic Acetylcholine Receptor Subunit Composition at the Synapse

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2009 Jul 18

PMID 19609303

Citations 31

Authors

Ruta B Almedom

Jana F Liewald

Guillermina Hernando

Christian Schultheis

Diego Rayes

Jie Pan

Thorsten Schedletzky

Harald Hutter

Cecilia Bouzat

Alexander Gottschalk

Affiliations

Soon will be listed here.

Abstract

Nicotinic acetylcholine receptors (nAChRs) are homo- or heteropentameric ligand-gated ion channels mediating excitatory neurotransmission and muscle activation. Regulation of nAChR subunit assembly and transfer of correctly assembled pentamers to the cell surface is only partially understood. Here, we characterize an ER transmembrane (TM) protein complex that influences nAChR cell-surface expression and functional properties in Caenorhabditis elegans muscle. Loss of either type I TM protein, NRA-2 or NRA-4 (nicotinic receptor associated), affects two different types of muscle nAChRs and causes in vivo resistance to cholinergic agonists. Sensitivity to subtype-specific agonists of these nAChRs is altered differently, as demonstrated by whole-cell voltage-clamp of dissected adult muscle, when applying exogenous agonists or after photo-evoked, channelrhodopsin-2 (ChR2) mediated acetylcholine (ACh) release, as well as in single-channel recordings in cultured embryonic muscle. These data suggest that nAChRs desensitize faster in nra-2 mutants. Cell-surface expression of different subunits of the 'levamisole-sensitive' nAChR (L-AChR) is differentially affected in the absence of NRA-2 or NRA-4, suggesting that they control nAChR subunit composition or allow only certain receptor assemblies to leave the ER.

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References

Chen B, Liu Q, Ge Q, Xie J, Wang Z . UNC-1 regulates gap junctions important to locomotion in C. elegans. Curr Biol. 2007; 17(15):1334-9. PMC: 1976340. DOI: 10.1016/j.cub.2007.06.060. View

Gray P, Harrison C, Vale W . Cripto forms a complex with activin and type II activin receptors and can block activin signaling. Proc Natl Acad Sci U S A. 2003; 100(9):5193-8. PMC: 154321. DOI: 10.1073/pnas.0531290100. View

Haffner C, Dettmer U, Weiler T, Haass C . The Nicastrin-like protein Nicalin regulates assembly and stability of the Nicalin-nodal modulator (NOMO) membrane protein complex. J Biol Chem. 2007; 282(14):10632-8. DOI: 10.1074/jbc.M611033200. View

Rawson J, Lee M, Kennedy E, Selleck S . Drosophila neuromuscular synapse assembly and function require the TGF-beta type I receptor saxophone and the transcription factor Mad. J Neurobiol. 2003; 55(2):134-50. DOI: 10.1002/neu.10189. View

Roberts B, Clucas C, Johnstone I . Loss of SEC-23 in Caenorhabditis elegans causes defects in oogenesis, morphogenesis, and extracellular matrix secretion. Mol Biol Cell. 2003; 14(11):4414-26. PMC: 266761. DOI: 10.1091/mbc.e03-03-0162. View

Fire A . Integrative transformation of Caenorhabditis elegans. EMBO J. 1986; 5(10):2673-80. PMC: 1167168. DOI: 10.1002/j.1460-2075.1986.tb04550.x. View

Shah S, Lee S, Tabuchi K, Hao Y, Yu C, LaPlant Q . Nicastrin functions as a gamma-secretase-substrate receptor. Cell. 2005; 122(3):435-47. DOI: 10.1016/j.cell.2005.05.022. View

Gu Y, Camacho P, Gardner P, Hall Z . Identification of two amino acid residues in the epsilon subunit that promote mammalian muscle acetylcholine receptor assembly in COS cells. Neuron. 1991; 6(6):879-87. DOI: 10.1016/0896-6273(91)90228-r. View

Culetto E, Baylis H, Richmond J, Jones A, Fleming J, Squire M . The Caenorhabditis elegans unc-63 gene encodes a levamisole-sensitive nicotinic acetylcholine receptor alpha subunit. J Biol Chem. 2004; 279(41):42476-83. DOI: 10.1074/jbc.M404370200. View

10.

Liewald J, Brauner M, Stephens G, Bouhours M, Schultheis C, Zhen M . Optogenetic analysis of synaptic function. Nat Methods. 2008; 5(10):895-902. DOI: 10.1038/nmeth.1252. View

11.

Shyu Y, Hiatt S, Duren H, Ellis R, Kerppola T, Hu C . Visualization of protein interactions in living Caenorhabditis elegans using bimolecular fluorescence complementation analysis. Nat Protoc. 2008; 3(4):588-96. DOI: 10.1038/nprot.2008.16. View

12.

Vashlishan A, Madison J, Dybbs M, Bai J, Sieburth D, Chng Q . An RNAi screen identifies genes that regulate GABA synapses. Neuron. 2008; 58(3):346-61. DOI: 10.1016/j.neuron.2008.02.019. View

13.

Yu G, Nishimura M, Arawaka S, Levitan D, Zhang L, Tandon A . Nicastrin modulates presenilin-mediated notch/glp-1 signal transduction and betaAPP processing. Nature. 2000; 407(6800):48-54. DOI: 10.1038/35024009. View

14.

Ren P, Lim C, Johnsen R, Albert P, Pilgrim D, Riddle D . Control of C. elegans larval development by neuronal expression of a TGF-beta homolog. Science. 1996; 274(5291):1389-91. DOI: 10.1126/science.274.5291.1389. View

15.

Keller S, Taylor P . Determinants responsible for assembly of the nicotinic acetylcholine receptor. J Gen Physiol. 1999; 113(2):171-6. PMC: 2223362. DOI: 10.1085/jgp.113.2.171. View

16.

Christensen M, Estevez A, Yin X, Fox R, Morrison R, McDonnell M . A primary culture system for functional analysis of C. elegans neurons and muscle cells. Neuron. 2002; 33(4):503-14. DOI: 10.1016/s0896-6273(02)00591-3. View

17.

Treinin M, Gillo B, LIEBMAN L, Chalfie M . Two functionally dependent acetylcholine subunits are encoded in a single Caenorhabditis elegans operon. Proc Natl Acad Sci U S A. 1998; 95(26):15492-5. PMC: 28070. DOI: 10.1073/pnas.95.26.15492. View

18.

Karlin A . Emerging structure of the nicotinic acetylcholine receptors. Nat Rev Neurosci. 2002; 3(2):102-14. DOI: 10.1038/nrn731. View

19.

Boulin T, Gielen M, Richmond J, Williams D, Paoletti P, Bessereau J . Eight genes are required for functional reconstitution of the Caenorhabditis elegans levamisole-sensitive acetylcholine receptor. Proc Natl Acad Sci U S A. 2008; 105(47):18590-5. PMC: 2587545. DOI: 10.1073/pnas.0806933105. View

20.

Eimer S, Gottschalk A, Hengartner M, Horvitz H, Richmond J, Schafer W . Regulation of nicotinic receptor trafficking by the transmembrane Golgi protein UNC-50. EMBO J. 2007; 26(20):4313-23. PMC: 2034668. DOI: 10.1038/sj.emboj.7601858. View