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Yeast two-hybrid screening identifies MPZ-1 and PTP-1 as candidate scaffolding proteins of metabotropic glutamate receptors in Caenorhabditis elegans

Yeast two-hybrid screening identifies MPZ-1 and PTP-1 as candidate scaffolding proteins of metabotropic glutamate receptors in Caenorhabditis elegans
Yeast two-hybrid screening identifies MPZ-1 and PTP-1 as candidate scaffolding proteins of metabotropic glutamate receptors in Caenorhabditis elegans

The metabotropic glutamate receptors (mGluRs) are a class of G-protein-coupled receptor that undergo extensive interactions with scaffolding proteins, and this is intrinsic to their function as an important group of neuromodulators at glutamatergic synapses. The Caenorhabditis elegans nervous system expresses three metabotropic glutamate receptors, MGL-1, MGL-2 and MGL-3. Relatively little is known about how the function and signalling of these receptors is organised in C. elegans. To identify proteins that scaffold the MGL-1 receptor, we have conducted a yeast two-hybrid screen. Three of the interacting proteins, MPZ-1, NRFL-1 and PTP-1, displayed motifs characteristic of mammalian mGluR scaffolding proteins. Using cellular co-expression criterion, we show mpz-1 and ptp-1 exhibited overlapping expression patterns with subsets of mgl-1 neurons. This included neurones in the pharyngeal nervous system that control the feeding organ of the worm. The mGluR agonist L-CCG-I inhibits the activity of this network in wild-type worms, in an MGL-1 and dose-dependent manner. We utilised L-CCG-I to identify if MGL-1 function was disrupted in mutants with deletions in the mpz-1 gene. The mpz-1 mutants displayed a largely wild-type response to L-CCG-I, suggesting MGL-1 signalling is not overtly disrupted consistent with a non-obligatory modulatory function in receptor scaffolding. The selectivity of the protein interactions and overlapping expression identified here warrant further investigation of the functional significance of scaffolding of metabotropic glutamate receptor function.

Caenorhabditis elegans, Electropharyngeogram, Glutamate, mGluR, Scaffolding
1354-2516
1-12
Dillon, James
f406e30a-3ad4-4a53-80db-6694bab5e3ed
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
O’Connor, Vincent
3c336b62-f3ef-4f78-a44c-ddc7b47bac3f
Dillon, James
f406e30a-3ad4-4a53-80db-6694bab5e3ed
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
O’Connor, Vincent
3c336b62-f3ef-4f78-a44c-ddc7b47bac3f

Dillon, James, Holden-Dye, Lindy and O’Connor, Vincent (2018) Yeast two-hybrid screening identifies MPZ-1 and PTP-1 as candidate scaffolding proteins of metabotropic glutamate receptors in Caenorhabditis elegans. Invertebrate Neuroscience, 18 (4), 1-12. (doi:10.1007/s10158-018-0218-2).

Record type: Article

Abstract

The metabotropic glutamate receptors (mGluRs) are a class of G-protein-coupled receptor that undergo extensive interactions with scaffolding proteins, and this is intrinsic to their function as an important group of neuromodulators at glutamatergic synapses. The Caenorhabditis elegans nervous system expresses three metabotropic glutamate receptors, MGL-1, MGL-2 and MGL-3. Relatively little is known about how the function and signalling of these receptors is organised in C. elegans. To identify proteins that scaffold the MGL-1 receptor, we have conducted a yeast two-hybrid screen. Three of the interacting proteins, MPZ-1, NRFL-1 and PTP-1, displayed motifs characteristic of mammalian mGluR scaffolding proteins. Using cellular co-expression criterion, we show mpz-1 and ptp-1 exhibited overlapping expression patterns with subsets of mgl-1 neurons. This included neurones in the pharyngeal nervous system that control the feeding organ of the worm. The mGluR agonist L-CCG-I inhibits the activity of this network in wild-type worms, in an MGL-1 and dose-dependent manner. We utilised L-CCG-I to identify if MGL-1 function was disrupted in mutants with deletions in the mpz-1 gene. The mpz-1 mutants displayed a largely wild-type response to L-CCG-I, suggesting MGL-1 signalling is not overtly disrupted consistent with a non-obligatory modulatory function in receptor scaffolding. The selectivity of the protein interactions and overlapping expression identified here warrant further investigation of the functional significance of scaffolding of metabotropic glutamate receptor function.

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Accepted/In Press date: 19 October 2018
e-pub ahead of print date: 11 November 2018
Published date: 1 December 2018
Keywords: Caenorhabditis elegans, Electropharyngeogram, Glutamate, mGluR, Scaffolding

Identifiers

Local EPrints ID: 426262
URI: https://eprints.soton.ac.uk/id/eprint/426262
ISSN: 1354-2516
PURE UUID: 7ca99319-09cf-4cf0-8f02-e2dacb4960af
ORCID for Lindy Holden-Dye: ORCID iD orcid.org/0000-0002-9704-1217

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Date deposited: 21 Nov 2018 17:30
Last modified: 10 Oct 2019 00:40

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Author: James Dillon
Author: Vincent O’Connor

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