Specific Activation of the G Protein-coupled Receptor BNGR-A21 by the Neuropeptide Corazonin from the Silkworm, Bombyx mori, Dually Couples to the Gq and Gs Signaling Cascades

Abstract

Background: The corazonin system is of importance for insects to control multiple physiological processes.Results: Bombyx BNGR-A21 was activated in signaling and internalization by direct interaction with corazonin.Conclusion: Bombyx corazonin was a specific ligand for dual Gq- and Gs-coupled BmCrzR.Significance: Our findings will lead to a better understanding of the corazonin system in regulation of fundamental physiological processes.

Identification and functional characterization of the pheromone biosynthesis activating neuropeptide receptor isoforms from Mamestra brassicae

“…To assess the role of arrestins in the internalization of the Mambr-PBANR isoforms, we constructed a fluorescent chimera of the non-visual Drosophila arrestin, Kurtz (DromeKurtz), fused at its C-terminus with the red fluorescent protein, mCherry, and co-transfected it with the respective PBANR variants. Consistent with previous reports (Yang et al, 2013(Yang et al, , 2016, DromeKurtz-mCherry localization was cytosolic in the absence of ligand stimulation (data not shown). Incubation with 1 μM Mambr-PT induced partial translocation of the Kurtz construct to the cell surface and/or co-localization with internalized receptors in cells expressing the Mambr-PBANR-B and -C isoforms (Fig.…”

“…This regulatory mechanism thus modulates the duration and intensity of the re ceptor-mediated signal transduction cascade (Hanyaloglu and Zastrow, 2008). Although ligand-induced internalization has been extensively studied in mammalian systems, the GPCRs that have been similarly characterized in insects are limited to the B. mori adipokinetic hormone (AKH) receptor (Huang et al, 2011), the B. mori corazonin receptor (Yang et al, 2013(Yang et al, , 2016, the Drosophila SP receptor (Hull and Brent, 2014), and three moth PBANRs (Hull et al, 2004(Hull et al, , 2005(Hull et al, , 2011Kawai et al, 2014;Lee et al, 2012). In each study, the respective receptor exhibited rapid internalization in mammalian and/ or insect cell lines following addition of labeled and unlabeled cognate ligands.…”

“…The non-visual Drosophila arrestin homolog, Kurtz, which exhibits significant sequence similarity with mammalian arrestins, functions similar to β-arrestin2 in receptor regulation, and as such has been useful in the characterization of insect GPCR internalization (Yang et al, 2013(Yang et al, , 2016. For arrestin translocation assays, a fluorescent chimera of Kurtz C-terminally tagged with the red fluorescent protein mCherry was constructed by overlap extension PCR (Wurch et al, 1998) using a cDNA clone (FBcl0171107; Drosophila Genomics Resource Center, Bloomington, IN) with chimeric primers (sense: 5′-GCTGAAACAGAGGCCATGGTGAGCAAGGGCGAG-3′; antisense: 5′-CTCGCCCTTGCTCACCATGGCCTCTGTTTCAGC-3′) and gene specific primers (sense Drosophila Kurtz: 5′-CATAAT-GAACGGTGGTGGTG-3′; antisense mCherry: 5′-CTACTTG-TACAGCTCGTC).…”

Structural basis for the interaction of diapause hormone with its receptor in the silkworm, Bombyx mori

“…As shown in Supplemental Table 1, all primers were designed on the basis of the sequence of GenBank Accession KP228012. For expression in Sf21 cells, corresponding PCR products were cloned into pBmIE1-Flag, which replaced the corresponding sites of pCMV-Flag, as previously described (11,20). The corresponding PCR products of Bommo-DHR and Bommo-DHR mutants were cloned into the HindIII and BamHI sites of pCMV-Flag/pBmIE1-Flag by using the Rapid DNA Ligation Kit (Beyotime), and we named these recombinant plasmids Flag-DHR and BmFlag-DHR, respectively.…”

Identification and Expression Profile of the Gonadotropin-Releasing Hormone Receptor in Common Chinese Cuttlefish,Sepiella japonica

“…GnRHRs and GnRH-like superfamily receptors (CrzR, AKHR, ACPR) have been identified in many invertebrate species such as Branchiostoma floridae, Octopus vulgaris, Crassostrea gigas, Nasonia vitripennis, and the Bombyx mori (Rodet et al, 2005;Tello et al, 2005;Kanda et al, 2006;Tello and Sherwood, 2009;Hansen et al, 2010;Yang et al, 2013). Transcriptional quantification analysis showed a specific expression of the C. gigas GnRH-related receptor (CgGnRH-R) in both male and female gonads during the reproductive cycle, demonstrating the involvement of CgGnRH-R in the control of reproduction (Rodet et al, 2005).…”