Identification of G protein-coupled receptors for Drosophila PRXamide peptides, CCAP, corazonin, and AKH supports a theory of ligand-receptor coevolution

Abstract

G-protein coupled receptors (GPCRs) are ancient, ubiquitous sensors vital to environmental and physiological signaling throughout organismal life. With the publication of the Drosophila genome, numerous “orphan” GPCRs have become available for functional analysis. Here we characterize two groups of GPCRs predicted as receptors for peptides with a C-terminal amino acid sequence motif consisting of −PRXamide (PRXa). Assuming ligand-receptor coevolution, two alternative hypotheses were constructed and tested. The insect PRXa peptides are evolutionarily related to the vertebrate peptide neuromedin U (NMU), or are related to arginine vasopressin (AVP), both of which have PRXa motifs. Seven Drosophila GPCRs related to receptors for NMU and AVP were cloned and expressed in Xenopus oocytes for functional analysis. Four Drosophila GPCRs in the NMU group (CG11475, CG8795, CG9918, CG8784) are activated by insect PRXa pyrokinins, (−FXPRXamide), Cap2b-like peptides (−FPRXamide), or ecdysis triggering hormones (−PRXamide). Three Drosophila GPCRs in the vasopressin receptor group respond to crustacean cardioactive peptide (CCAP), corazonin, or adipokinetic hormone (AKH), none of which are PRXa peptides. These findings support a theory of coevolution for NMU and Drosophila PRXa peptides and their respective receptors

The pheromone biosynthesis activating neuropeptide (PBAN) receptor of Heliothis virescens: Identification, functional expression, and structure–activity relationships of ligand analogs

“…The PBANRclade is further supported by pharmacological profiles shared by its members: All receptors in this clade show marked sensitivity to pyrokinins/PBAN, which feature the FXPRLa Cterminal amino acid motif [3,9,21,26]. The PBANR clade is grouped with CG9918, mainly sensitive to CAP2b-3 and a diapause hormone (DH) analog that contains a Cterminal WFGPRLa motif [2,21]. Finally, the PBANR clade and CG9918 are grouped with CG14575, which is highly sensitive and selective to CAP2b-1 and CAP2b-2 peptides featuring the C-terminal sequence motif FPRVa ( Fig.…”

“…In the distance tree, three moth PBANRs (HevPBANR, HezPBANR and BomPBANR) are grouped with Drosophila GPCRs CG8784 and CG8795 to produce the monophyletic 'PBANR clade' with a 97% bootstrap value. The PBANRclade is further supported by pharmacological profiles shared by its members: All receptors in this clade show marked sensitivity to pyrokinins/PBAN, which feature the FXPRLa Cterminal amino acid motif [3,9,21,26]. The PBANR clade is grouped with CG9918, mainly sensitive to CAP2b-3 and a diapause hormone (DH) analog that contains a Cterminal WFGPRLa motif [2,21].…”

“…We and others showed previously that GPCRs CG8795 and CG8784 are highly sensitive to D. melanogaster pyrokinin-2 (DrmPK-2), a neuropeptide with high C-terminal sequence similarity to PBAN [21,26]. These findings suggested that the moth counterpart of CG8795 would likely encode a functional PBAN receptor.…”

“…In order to identify a PBAN receptor from Heliothis virescens, we synthesized degenerate PCR primers based on previously published sequences of Drosophila melanogaster GPCRs, CG8795 and CG8784, both of which are highly sensitive to DrmPK-2, a likely homolog of PBAN [21,26]. PCRs using nested sets of degenerate primers yielded a ~636 bp cDNA fragment from the CNS of 4 th instar larvae.…”

An obligatory role for neurotensin in high-fat-diet-induced obesity

“…8a, b). Among the three potential Drosophila NTRs 26,27 , RNAi of CG9918 (Pyrokinin 1 receptor, PK1-R), but not CG8784 or CG8795, blocked the decrease in p-AMPK levels in NT-expressing cells (Extended Data Fig. 8a).…”