title: Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea.
abstract: The diapause hormone (DH) in the heliothine moth has shown its activity in termination of pupal diapause, while the orthology in the silkworm is known to induce embryonic diapause. In the current study, we cloned the diapause hormone receptor from the corn earworm Helicoverpa zea (HzDHr) and tested its ligand specificities in a heterologous reporter system. HzDHr was expressed in Chinese Hamster Ovary (CHO) cells, which were co-transfected with the aequorin reporter, and was used to measure the ligand activities. A total of 68 chemicals, including natural DH analogs and structurally similar peptide mimetics, were tested for agonistic and antagonistic activities. Several peptide mimetics with a 2-amino-7-bromofluorene-succinoyl (2Abf-Suc) N-terminal modification showed strong agonistic activities; these mimetics included 2Abf-Suc-F[dA]PRLamide, 2Abf-Suc-F[dR]PRLamide, 2Abf-Suc-FKPRLamide and 2Abf-Suc-FGPRLamide. Antagonistic activity was found in the ecdysis triggering hormone in Drosophila melanogaster (FFLKITKNVPRLamide). Interestingly, HzDHr does not discriminate between DH (WFGPRLamide C-terminal motif) and another closely related endogenous peptide, pyrokinin 1 (FXPRXamide; a C-terminal motif that is separate from WFGPRLamide). We provide large-scale in vitro data that serve as a reference for the development of agonists and antagonists to disrupt the DH signaling pathway.
authors.affiliation: Department of Entomology, Kansas State University, Manhattan, KS 66506, United States.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Entomology, Kansas State University, Manhattan, KS 66506, United States.
authors.forename: Zhaojun
authors.initials: Z
authors.lastname: Wei
authors.affiliation: Areawide Pest Management Research, Southern Plains Agricultural Research Center, USDA, 2881 F/B Road, College Station, TX 77845, United States.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Nachman
authors.affiliation: Department of Entomology, Kansas State University, Manhattan, KS 66506, United States. Electronic address: ypark@ksu.edu.
authors.forename: Yoonseong
authors.initials: Y
authors.lastname: Park
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D019476:Insect Proteins / Q000737:chemistry / Q000378:metabolism*; D009036:Moths / Q000235:genetics / Q000378:metabolism*; D009479:Neuropeptides / Q000737:chemistry / Q000378:metabolism; D057786:Peptidomimetics; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D009479:Neuropeptides; D057786:Peptidomimetics; D043562:Receptors, G-Protein-Coupled
keywords: GPCR; Insect; Neuropeptide; Peptidomimetic