Primary structure of a developmentally regulated nicotinic acetylcholine receptor protein from <i>Drosophila</i>

Hermans-Borgmeyer, I; Zopf, Dieter; Ryseck, Rolf-Peter; Hovemann, Bernd; Betz, Heinrich; Gundelfinger, Eckart D.

doi:10.1002/j.1460-2075.1986.tb04389.x

Cited by 139 publications

(64 citation statements)

References 46 publications

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“…The hygromycin selection plasmid, pCOHygro (van der Straten et al, 1989), was provided by Dr. Martin Rosenberg (SmithKline Beecham Pharmaceuticals, Philadelphia, PA, U.S.A.). The Drosophila nAChR subunit cDNAs SAD, ALS, and ARD in plasmid pBluescript have been described previously (Hermans-Borgmeyer et al, 1986;Bossy et al, 1988;Sawruk et al, 1990a). The four Drosophila subunit cDNAs were subcloned into pRmHa3 and the mammalian expression vector, pcDNA3 (Invitrogen).…”

Section: Plasmids and Cell Linesmentioning

confidence: 99%

“…In addition, some vertebrate neuronal nAChR subunits (a7, a8, and a9) have been shown to form functional homooligomeric channels (Couturier et al, 1990;Elgoyhen et al, 1994). Of the four nAChR subunit cDNAs (ALS, ARD, SAD, and SBD) cloned from Drosophila (Hermans-Borgmeyer et al, 1986;Bossy et al, 1988;Sawruk et al, 1990a,b), ALS and SAD appear to be "a-like," because they contain residues believed to contribute to the ACh-binding site. However, in contrast to the vertebrate muscle and neuronal nAChRs, the expression of functional nAChRs from these Drosophila cDNAs in Xenopus oocytes has proved to be difficult (Gundelfinger, 1992;Gundelfinger and Hess, 1992).…”

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confidence: 99%

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Temperature‐Sensitive Expression of Drosophila Neuronal Nicotinic Acetylcholine Receptors

Lansdell

Schmitt

Betz

et al. 1997

Journal of Neurochemistry

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Heterologous expression of cloned Drosophila nicotinic acetylcholine receptor (nAChR) subunits indicates that these proteins misfold when expressed in mammalian cell lines at 37°C.This misfolding can, however, be overcome either by growing transfected mammalian cells at lower temperatures or by the expression of Drosophila nAChR subunits in a Drosophila cell line. Whereas the Drosophila nAChR /3 subunit (SBD) cDNA, reported previously, lacked part of the SBD coding sequence, here we report the construction and expression of a full-length SBD cDNA. We have examined whether problems in expressing functional Drosophila nAChRs in either Xenopus oocytes or mammalian cell lines can be attributed to an inability of these expression systems to assemble correctly Drosophila nAChRs. Despite expression in what might be considered a more native cellular environment, we have been unable to detect functional nAChRs in a Drosophila cell line unless Drosophila nAChR subunit cDNA5 are coexpressed with vertebrate nAChR subunits. Our results indicate that the folding of Drosophila nAChR subunits is temperature-sensitive and strongly suggest that the inability of these Drosophila nAChR subunits to generate functional channels in the absence of vertebrate subunits is due to a requirement for coassembly with as yet unidentified Drosophila nAChR subunits. Key Words: Protein folding -Assembly-Nicotinic acetylcholine receptor.

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Section: Plasmids and Cell Linesmentioning

confidence: 99%

mentioning

confidence: 99%

Temperature‐Sensitive Expression of Drosophila Neuronal Nicotinic Acetylcholine Receptors

Lansdell

Schmitt

Betz

et al. 1997

Journal of Neurochemistry

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120

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“…The sequence of the Drosophila genome predicts seven ␣-and three non-␣-subunits. The characterization of four of the ␣-subunits (ALS, SAD, D␣3, and D␣4) (8 -10) as well as two of the ␤-subunits (ARD and SBD) (11,12) was previously carried out through screening with homologous probes of vertebrate nAChR cDNAs. The existence of the remaining three ␣-subunits D␣5 (13,14) and D␣6 and D␣7) (14) as well as D␤3 (15) was recently confirmed by reverse transcription-PCR based on the Drosophila genome information (16).…”

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confidence: 99%

The Drosophila Acetylcholine Receptor Subunit Dα5 Is Part of an α-Bungarotoxin Binding Acetylcholine Receptor

Wu¹,

Ma²,

Pierzchala

et al. 2005

Journal of Biological Chemistry

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The central nervous system of Drosophila melanogaster contains an ␣-bungarotoxin-binding protein with the properties expected of a nicotinic acetylcholine receptor. This protein was purified 5800-fold from membranes prepared from Drosophila heads. The protein was solubilized with 1% Triton X-100 and 0.5 M sodium chloride and then purified using an ␣-cobratoxin column followed by a lentil lectin affinity column. The purified protein had a specific activity of 3.9 mol of 125 I-␣-bungarotoxin binding sites/g of protein.The subunit composition of the purified receptor was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. This subunit profile was identical with that revealed by in situ labeling of the membrane-bound protein using the photolyzable methyl Nicotinic acetylcholine receptors (nAChRs)1 are key elements of fast cholinergic synaptic transmission (1, 2). They are part of a large superfamily of ligand-gated ion channels comprising 5HT 3 , GABA A , and GABA C , glycine, and invertebrate anionic glutamate receptors. nAChRs are widely distributed in both vertebrates and invertebrates. In addition to their occurrence in the highly specialized tissue of the electrical organ of certain species of fish (Torpedo californica, Electrophorus electricus), nAChRs are mostly found at the vertebrate neuromuscular junction and the nervous system of both vertebrates and invertebrates (3, 4).Structurally, nAChRs are formed by assembly of their subunits into a pentameric membrane protein complex. Subunits carrying two conserved adjacent cysteines in the amino-terminal domain in the vicinity of the acetylcholine binding site are classified as ␣-subunits, while the remainder of the subunit types are designated non-␣ or -␤. The individual subunits have a large amino-terminal domain, which includes the binding site for the ligand, followed by four membrane spanning domains M1-M4. The M2 segments of the five subunits form the lining of the ion channel, which is transiently permeable to cations (Na ϩ , K ϩ , Ca 2ϩ ) upon binding of the ligand acetylcholine (ACh). Between M3 and M4 a large intracellular loop exists, which contains several phosphorylation sites required for receptor stabilization (for reviews, see Refs. 3, 5, and 6).Biochemical analysis of receptors subunits composition in a variety of species as well the information obtained from the sequencing of the genome of humans, mice, the nematode Caenorhabditis elegans and the fruit fly Drosophila melanogaster indicates a large number of genes encoding nicotinic acetylcholine receptor subunits. Nicotinic acetylcholine receptors from muscle have an invariant subunit composition ((␣1) 2 (␤1)␦␥ (fetal) or ⑀). Neuronal nicotinic acetylcholine receptors are composed of ␣ and non-␣-subunits with a stoichoimetry of (␣) 2 (non-␣) 3 . The large number of genes encoding nAChR subunits in the nervous system can give rise to an array of functionally distinct nAChRs. Since the vertebrate nervous system also contains homomeric nAChRs (e.g. (␣ 7 ) 5 ) the entire range ...

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“…The time course of the developmental expression of the Dor2 gene shows a remarkable coincidence with that of the ARD gene [19,21,31]; it differs, however, significantly from the expression profile of the ALS Fig. 3.…”

Section: Developmental Expression Of the Da2 Genementioning

confidence: 80%

Structure and developmental expression of the Dα2 gene encoding a novel nicotinic acetylcholine receptor protein of Drosophila melanogaster

et al. 1990

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Nicotinic acetylcholine receptors (nAChRs) represent a heterogeneous group of excitatory neurotransmitter receptors in the insect brain. We have characterized the Da2 gene of Drosophila melanoguster, a new member of the nAChR gene family. The protein coding region is interrupted by six introns. The positions of three of these introns are shared with all other nAChR genes. The deduced Da2 protein shows the structural features of ligand-binding nAChR a-subunits. Cytogenetically, the Da2 gene maps at position 96A of the 3rd chromosome, close to the ALS gene which also encodes an a-like nAChR subunit. Da2 transcripts are predominantly expressed in late embryos and in fly heads.

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Primary structure of a developmentally regulated nicotinic acetylcholine receptor protein from Drosophila

Cited by 139 publications

References 46 publications

Temperature‐Sensitive Expression of Drosophila Neuronal Nicotinic Acetylcholine Receptors

Temperature‐Sensitive Expression of Drosophila Neuronal Nicotinic Acetylcholine Receptors

The Drosophila Acetylcholine Receptor Subunit Dα5 Is Part of an α-Bungarotoxin Binding Acetylcholine Receptor

Structure and developmental expression of the Dα2 gene encoding a novel nicotinic acetylcholine receptor protein of Drosophila melanogaster

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