RRC ID 3383
著者 Gottschalk A, Almedom RB, Schedletzky T, Anderson SD, Yates JR 3rd, Schafer WR.
タイトル Identification and characterization of novel nicotinic receptor-associated proteins in Caenorhabditis elegans.
ジャーナル EMBO J
Abstract 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.
巻・号 24(14)
ページ 2566-78
公開日 2005-7-20
DOI 10.1038/sj.emboj.7600741
PII 7600741
PMID 15990870
PMC PMC1176467
MeSH Affinity Labels Animals Caenorhabditis elegans Proteins / metabolism* Carrier Proteins / metabolism Genes, Reporter Levamisole / metabolism* Muscles / drug effects Neurons / drug effects Nicotine / pharmacology Nicotinic Agonists / pharmacology RNA Interference Receptors, Cholinergic / chemistry Receptors, Cholinergic / metabolism Receptors, Fibroblast Growth Factor / metabolism Receptors, Nicotinic / chemistry Receptors, Nicotinic / metabolism* Synapses / metabolism Time Factors
IF 9.889
引用数 100
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
リソース情報
線虫 tm1395 tm1453 tm1649