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Article

  • The EMBO Journal (2005) 24, 2566 - 2578
  • doi:10.1038/sj.emboj.7600741

Published online: 30 June 2005

Identification and characterization of novel nicotinic receptor-associated proteins in Caenorhabditis elegans

Alexander Gottschalk1,2, Ruta B Almedom1, Thorsten Schedletzky1, Scott D Anderson3, John R Yates III3 and William R Schafer2

  1. Institute for Biochemistry, Goethe-University, Frankfurt, Germany
  2. Division of Biological Sciences, University of California, San Diego, La Jolla, CA, USA
  3. Department of Cell Biology, The Scripps Research Institute, La Jolla, CA, USA

Correspondence to:

William R Schafer, Division of Biological Sciences, University of California, San Diego, 9500 Gilman Dr, La Jolla, CA 92093-0643, USA. Tel.: +1 858 822 0508; Fax: +1 858 822 2003; E-mail: wschafer@ucsd.edu

Alexander Gottschalk, Institute for Biochemistry, Goethe-University, Biocenter N210, Marie-Curie-Str. 9, 60439 Frankfurt, Germany. Tel.: +49 69 798 29261; Fax: +49 69 798 29495; E-mail: a.gottschalk@em.uni-frankfurt.de

Received 3 December 2004; Accepted 13 June 2005

Nicotinic acetylcholine receptors (nAChRs) mediate fast excitatory neurotransmission in neurons and muscles. To identify nAChR accessory proteins, which may regulate their expression or function, we performed tandem affinity purification of the levamisole-sensitive nAChR from Caenorhabditis elegans, mass spectrometry of associated components, and RNAi-based screening for effects on in vivo nicotine sensitivity. Among the proteins identified was the calcineurin A subunit TAX-6, which appeared to function as a negative regulator of nAChR activity. We also identified five proteins not previously linked to nAChR function, whose inactivation conferred nicotine resistance, implicating them as positive regulators of nAChR activity. Of these, the copine NRA-1 colocalized with the levamisole receptor at neuronal and muscle plasma membranes, and, when mutated, caused reduced synaptic nAChR expression. Loss of SOC-1, which acts in receptor tyrosine kinase (RTK) signaling, also reduced synaptic levamisole receptor levels, as did mutations in the fibroblast growth factor receptor EGL-15, and another RTK, CAM-1. Thus, tandem affinity purification is a viable approach to identify novel proteins regulating neurotransmitter receptor activity or expression in model systems like C. elegans.

  • Keywords:

    • copine,
    • levamisole receptor,
    • SOC-1,
    • tandem affinity purification,
    • TAX-6 calcineurin