Cloning and functional characterization of a Caenorhabditis elegans muscarinic acetylcholine receptor.
A cDNA clone encoding a muscarinic acetylcholine receptor (mAChR) has been isolated from the nematode Caenorhabditis elegans. The nematode mAChR, consisted of 585 amino acids, displays a high degree of amino acid sequence homology to other invertebrate and vertebrate mAChRs. Excluding a highly variable middle portion of the third intracellular loop, the C. elegans mAChR shares about 51% amino acid sequence identity with a Drosophila mAChR and 42-44% identity with human m1-m5 mAChR subtypes. Comparison of the cDNA sequence with the corresponding genomic sequence reveals that the C. elegans mAChR gene contains ten introns, eight of them in the coding region. Pharmacological profiles of the C. elegans mAChR expressed in Chinese hamster ovary (CHO) cells were shown to be similar to those of mammalian counterparts, indicating that ligand binding domains of the receptor have been conserved during evolution. When this cloned receptor was expressed in Xenopus oocytes, acetylcholine evoked a transient Cl- current. Furthermore, activation of the receptor with oxotremorine, acetylcholine or carbachol resulted in the stimulation of phosphatidylinositol metabolism in CHO cells, suggesting that the receptor is coupled to phospholipase C activation.
Hwang
J M
JM
Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Cheongju, South Korea.
Chang
D J
DJ
Kim
U S
US
Lee
Y S
YS
Park
Y S
YS
Kaang
B K
BK
Cho
N J
NJ
D000109:Acetylcholine; D000595:Amino Acid Sequence; D000818:Animals; D001285:Atropine / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive; D016466:CHO Cells; D017173:Caenorhabditis elegans / Q000235:genetics*; D018118:Chloride Channels / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D015870:Gene Expression; D015640:Ion Channel Gating; D008024:Ligands; D008969:Molecular Sequence Data; D019832:N-Methylscopolamine / Q000378:metabolism; D010095:Oxotremorine / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012601:Scopolamine / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014981:Xenopus
D018118:Chloride Channels; D008024:Ligands; D011976:Receptors, Muscarinic; D011994:Recombinant Proteins; D010095:Oxotremorine; D001285:Atropine; D012601:Scopolamine; D000109:Acetylcholine; D019832:N-Methylscopolamine