Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans.
The unc-17 gene encodes the vesicular acetylcholine transporter (VAChT) in Caenorhabditis elegans. unc-17 reduction-of-function mutants are small, slow growing, and uncoordinated. Several independent unc-17 alleles are associated with a glycine-to-arginine substitution (G347R), which introduces a po...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
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2012
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| _version_ | 1797092428187959296 |
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| author | Mathews, E Mullen, G Hodgkin, J Duerr, J Rand, J |
| author_facet | Mathews, E Mullen, G Hodgkin, J Duerr, J Rand, J |
| author_sort | Mathews, E |
| collection | OXFORD |
| description | The unc-17 gene encodes the vesicular acetylcholine transporter (VAChT) in Caenorhabditis elegans. unc-17 reduction-of-function mutants are small, slow growing, and uncoordinated. Several independent unc-17 alleles are associated with a glycine-to-arginine substitution (G347R), which introduces a positive charge in the ninth transmembrane domain (TMD) of UNC-17. To identify proteins that interact with UNC-17/VAChT, we screened for mutations that suppress the uncoordinated phenotype of UNC-17(G347R) mutants. We identified several dominant allele-specific suppressors, including mutations in the sup-1 locus. The sup-1 gene encodes a single-pass transmembrane protein that is expressed in a subset of neurons and in body muscles. Two independent suppressor alleles of sup-1 are associated with a glycine-to-glutamic acid substitution (G84E), resulting in a negative charge in the SUP-1 TMD. A sup-1 null mutant has no obvious deficits in cholinergic neurotransmission and does not suppress unc-17 mutant phenotypes. Bimolecular fluorescence complementation (BiFC) analysis demonstrated close association of SUP-1 and UNC-17 in synapse-rich regions of the cholinergic nervous system, including the nerve ring and dorsal nerve cords. These observations suggest that UNC-17 and SUP-1 are in close proximity at synapses. We propose that electrostatic interactions between the UNC-17(G347R) and SUP-1(G84E) TMDs alter the conformation of the mutant UNC-17 protein, thereby restoring UNC-17 function; this is similar to the interaction between UNC-17/VAChT and synaptobrevin. |
| first_indexed | 2024-03-07T03:45:47Z |
| format | Journal article |
| id | oxford-uuid:bf698720-3b6b-4220-8651-def2b95d0666 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:45:47Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | oxford-uuid:bf698720-3b6b-4220-8651-def2b95d06662022-03-27T05:47:16ZGenetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bf698720-3b6b-4220-8651-def2b95d0666EnglishSymplectic Elements at Oxford2012Mathews, EMullen, GHodgkin, JDuerr, JRand, JThe unc-17 gene encodes the vesicular acetylcholine transporter (VAChT) in Caenorhabditis elegans. unc-17 reduction-of-function mutants are small, slow growing, and uncoordinated. Several independent unc-17 alleles are associated with a glycine-to-arginine substitution (G347R), which introduces a positive charge in the ninth transmembrane domain (TMD) of UNC-17. To identify proteins that interact with UNC-17/VAChT, we screened for mutations that suppress the uncoordinated phenotype of UNC-17(G347R) mutants. We identified several dominant allele-specific suppressors, including mutations in the sup-1 locus. The sup-1 gene encodes a single-pass transmembrane protein that is expressed in a subset of neurons and in body muscles. Two independent suppressor alleles of sup-1 are associated with a glycine-to-glutamic acid substitution (G84E), resulting in a negative charge in the SUP-1 TMD. A sup-1 null mutant has no obvious deficits in cholinergic neurotransmission and does not suppress unc-17 mutant phenotypes. Bimolecular fluorescence complementation (BiFC) analysis demonstrated close association of SUP-1 and UNC-17 in synapse-rich regions of the cholinergic nervous system, including the nerve ring and dorsal nerve cords. These observations suggest that UNC-17 and SUP-1 are in close proximity at synapses. We propose that electrostatic interactions between the UNC-17(G347R) and SUP-1(G84E) TMDs alter the conformation of the mutant UNC-17 protein, thereby restoring UNC-17 function; this is similar to the interaction between UNC-17/VAChT and synaptobrevin. |
| spellingShingle | Mathews, E Mullen, G Hodgkin, J Duerr, J Rand, J Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title | Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title_full | Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title_fullStr | Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title_full_unstemmed | Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title_short | Genetic interactions between UNC-17/VAChT and a novel transmembrane protein in Caenorhabditis elegans. |
| title_sort | genetic interactions between unc 17 vacht and a novel transmembrane protein in caenorhabditis elegans |
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