| Title: | Identification and functional characterization of the pheromone biosynthesis activating neuropeptide receptor isoforms from Mamestra brassicae |
| Author(s): | Fodor J; Hull JJ; Koblos G; Jacquin-Joly E; Szlanka T; Fonagy A; |
| Address: | "Plant Protection Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, H-1022 Budapest, Hungary. Agricultural Research Service, United States Department of Agriculture, Arid Land Agricultural Research Center, Maricopa, AZ, USA. Plant Protection Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, H-1022 Budapest, Hungary. Electronic address: koblos.gabriella@agrar.mta.hu. INRA iEES-Paris, Institute of Ecology and Environmental Sciences, Route de Saint-Cyr, Cedex 78026 Versailles, France. Institute of Biochemistry, Biological Research Centre, Hungarian Academy of Sciences, H-6726 Szeged, Hungary" |
| DOI: | 10.1016/j.ygcen.2017.05.024 |
| ISSN/ISBN: | 1095-6840 (Electronic) 0016-6480 (Linking) |
| Abstract: | "In most moth species, including Mamestra brassicae, pheromone biosynthesis activating neuropeptide (PBAN) regulates pheromone production. Generally, PBAN acts directly on the pheromone gland (PG) cells via its specific G protein-coupled receptor (i.e. PBANR) with Ca(2+) as a second messenger. In this study, we identified cDNAs encoding three variants (A, B and C) of the M. brassicae PBANR (Mambr-PBANR). The full-length coding sequences were transiently expressed in cultured Trichoplusia ni cells and Sf9 cells for functional characterization. All three isoforms dose-dependently mobilized extracellular Ca(2+) in response to PBAN analogs with Mambr-PBANR-C exhibiting the greatest sensitivity. Fluorescent confocal microscopy imaging studies demonstrated binding of a rhodamine red-labeled ligand (RR10CPBAN) to all three Mambr-PBANR isoforms. RR10CPBAN binding did not trigger ligand-induced internalization in cells expressing PBANR-A, but did in cells expressing the PBANR-B and -C isoforms. Furthermore, activation of the PBANR-B and -C isoforms with the 18 amino acid Mambr-pheromonotropin resulted in co-localization with a Drosophila melanogaster arrestin homolog (Kurtz), whereas stimulation with an unrelated peptide had no effect. PCR-based profiling of the three transcripts revealed a basal level of expression throughout development with a dramatic increase in PG transcripts from the day of adult emergence with PBANR-C being the most abundant" |
| Keywords: | "Amino Acid Sequence Animals Cells, Cultured Cloning, Molecular DNA, Complementary/genetics Drosophila Proteins/chemistry/metabolism Drosophila melanogaster/metabolism Endocytosis Female Gene Expression Profiling Ligands Moths/genetics/*metabolism Neuropep;" |
| Notes: | "MedlineFodor, Jozsef Hull, J Joe Koblos, Gabriella Jacquin-Joly, Emmanuelle Szlanka, Tamas Fonagy, Adrien eng Research Support, Non-U.S. Gov't 2017/06/06 Gen Comp Endocrinol. 2018 Mar 1; 258:60-69. doi: 10.1016/j.ygcen.2017.05.024. Epub 2017 Jun 1" |
