Cloning and characterization of the pheromone biosynthesis activating neuropeptide receptor gene in Spodoptera littoralis larvae.
In noctuid moths cuticular pigmentation is regulated by the pyrokinin/pheromone biosynthesis activating neuropeptide (PK/PBAN) family, which also mediates a variety of other functions in moths and other insects. Numerous studies have shown that these neuropeptides exert their functions through activation of the PBAN receptor (PBAN-R), with subsequent Ca(2+) influx, followed by either activation of cAMP or direct activation of downstream kinases. Recently, several PBAN-Rs have been identified, all of which are from the pheromone gland of adult female moths, but evidence shows that functional PK/PBAN-Rs can also be expressed in insect larvae, where they mediate melanization and possibly other functions (e.g., diapause). Here, we identified a gene encoding a G-protein-coupled receptor from the 5th instar larval tissue of the moth Spodoptera littoralis. The cDNA of this gene contains an open reading frame with a length of 1050 nucleotides, which translates to a 350-amino acid, 42-kDa protein that shares 92% amino acid identity with Helicoverpa zea and Helicoverpa armigera PBAN-R, 81% with Bombyx mori PBAN-R and 72% with Plutella xylostella PBAN-R. The S. littoralis PBAN-R gene was stably expressed in NIH3T3 cells and transiently in HEK293 cells. We show that it mediates the dose-dependent PBAN-induced intracellular Ca(2+) response and activation of the MAP kinase via a PKC-dependent but Galphai-independent signaling mechanism. Other PK/PBAN family peptides (pheromonotropin and a C-terminally PBAN-derived peptide PBAN(28-33)NH(2)) also triggered MAP kinase activation. This receptor, together with the previously cloned PBAN-R, may facilitate our understanding of the cell-specific responses and functional diversities of this diverse neuropeptide family.
Zheng
Lei
L
Department of Cell Biology and Neurosciences, University of California, Riverside, CA 92521, United States.
Lytle
Christian
C
Njauw
Ching-Ni
CN
Altstein
Miriam
M
Martins-Green
Manuela
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D020013:Calcium Signaling / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D007814:Larva / Q000187:drug effects; D020935:MAP Kinase Signaling System; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009479:Neuropeptides / Q000819:agonists / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D018013:Receptors, Neuropeptide / Q000737:chemistry / Q000235:genetics*; D018411:Spodoptera / Q000187:drug effects / Q000235:genetics*
D018076:DNA, Complementary; D009479:Neuropeptides; D010455:Peptides; D018013:Receptors, Neuropeptide; C060896:pheromone-biosynthesis-activating neuropeptide I