« Previous AbstractInfluence of water availability on gender determination of gametophytes in a diploid-polyploid complex of a xerophytic fern genus    Next AbstractCOVID-19 lockdown: Effects on selected volatile organic compound (VOC) emissions over the major Indian metro cities »

J Recept Signal Transduct Res

Title:		Engineered yeasts as reporter systems for odorant detection
Author(s):		Pajot-Augy E; Crowe M; Levasseur G; Salesse R; Connerton I;
Address:		"INRA, Biologie Cellulaire et Moleculaire, Equipe Recepteurs et Communication Cellulaire, Jouy-en-Josas, France. pajot@jouy.inra.fr"
Journal Title:		J Recept Signal Transduct Res
Year:		2003
Volume:		23
Issue:		2-Mar
Page Number:		155 - 171
DOI:		10.1081/rrs-120025196
ISSN/ISBN:		1079-9893 (Print) 1079-9893 (Linking)
Abstract:		"The functional expression of olfactory receptors (ORs) is a primary requirement to utilize olfactory detection systems. We have taken advantage of the functional similarities between signal transduction cascades in the budding yeast Saccharomyces cerevisiae and mammalian cells. The yeast pheromone response pathway has been adapted to allow ligand-dependent signaling of heterologous expressed G-protein coupled receptors (GPCRs) via mammalian or chimeric yeast/mammalian Galpha proteins. Two different strategies are reported here which offer a positive screen for functional pairs. The OR and Galpha protein are introduced into the modified yeast cells such that they hijack the pheromone response pathway usually resulting in cell cycle arrest. The first strategy utilizes ligand-induced expression of a FUS1-HIS3 reporter gene to permit growth on a selective medium lacking histidine; the second to induce ligand-dependent expression of a FUSI-Hph reporter gene, conferring resistance to hygromycin. Validation of the systems was performed using the rat 17 receptor response to a range of aldehyde odorants previously characterized as functional ligands. Of these only heptanal produced a positive growth response in the concentration range 5 x 10(-8) to 5 x 10(-6) M. Induction conditions appear to be critical for functional expression, and the solvents of odorants have a toxic effect for the highest odorant concentrations. The preference of rat 17 receptor for the ligand heptanal in yeast has to be compared to concurrent results obtained with mammalian expression systems"
Keywords:		"Animals Genes, Reporter Histidine/metabolism Ligands Olfactory Receptor Neurons/cytology/*metabolism Pheromones/metabolism Plasmids/genetics/metabolism Protein Subunits/genetics/metabolism Rats Receptors, Odorant/genetics/*metabolism Saccharomyces cerevis;"
Notes:		"MedlinePajot-Augy, Edith Crowe, Mark Levasseur, Gregoire Salesse, Roland Connerton, Ian eng England 2003/11/25 J Recept Signal Transduct Res. 2003; 23(2-3):155-71. doi: 10.1081/rrs-120025196"