| Title: | Functional coupling of a nematode chemoreceptor to the yeast pheromone response pathway |
| Author(s): | Tehseen M; Dumancic M; Briggs L; Wang J; Berna A; Anderson A; Trowell S; |
| Address: | "CSIRO Food Futures National Research Flagship & CSIRO Ecosystem Sciences, Australia, PO Box 1700, Canberra, ACT 2601, Australia" |
| DOI: | 10.1371/journal.pone.0111429 |
| ISSN/ISBN: | 1932-6203 (Electronic) 1932-6203 (Linking) |
| Abstract: | "Sequencing of the Caenorhabditis elegans genome revealed sequences encoding more than 1,000 G-protein coupled receptors, hundreds of which may respond to volatile organic ligands. To understand how the worm's simple olfactory system can sense its chemical environment there is a need to characterise a representative selection of these receptors but only very few receptors have been linked to a specific volatile ligand. We therefore set out to design a yeast expression system for assigning ligands to nematode chemoreceptors. We showed that while a model receptor ODR-10 binds to C. elegans Galpha subunits ODR-3 and GPA-3 it cannot bind to yeast Galpha. However, chimaeras between the nematode and yeast Galpha subunits bound to both ODR-10 and the yeast Gbetagamma subunits. FIG2 was shown to be a superior MAP-dependent promoter for reporter expression. We replaced the endogenous Galpha subunit (GPA1) of the Saccharomyces cerevisiae (ste2Delta sst2Delta far1Delta) triple mutant ('Cyb') with a Gpa1/ODR-3 chimaera and introduced ODR-10 as a model nematode GPCR. This strain showed concentration-dependent activation of the yeast MAP kinase pathway in the presence of diacetyl, the first time that the native form of a nematode chemoreceptor has been functionally expressed in yeast. This is an important step towards en masse de-orphaning of C. elegans chemoreceptors" |
| Keywords: | "Animals Caenorhabditis elegans/*metabolism Chemoreceptor Cells/*metabolism Chromosomes, Fungal Green Fluorescent Proteins/genetics Microscopy, Confocal Pheromones/*metabolism Plasmids Saccharomyces cerevisiae/genetics/*metabolism Saccharomyces cerevisiae;" |
| Notes: | "MedlineTehseen, Muhammad Dumancic, Mira Briggs, Lyndall Wang, Jian Berna, Amalia Anderson, Alisha Trowell, Stephen eng Research Support, Non-U.S. Gov't 2014/11/22 PLoS One. 2014 Nov 21; 9(11):e111429. doi: 10.1371/journal.pone.0111429. eCollection 2014" |
