Molecular identification and expression analysis of a diapause hormone receptor in the corn earworm, Helicoverpa zea

被引:11
作者
Zhang, Qirui [1 ,2 ]
Piermarini, Peter M. [3 ]
Nachman, Ronald J. [4 ]
Denlinger, David L. [1 ,2 ]
机构
[1] Ohio State Univ, Dept Entomol, Columbus, OH 43210 USA
[2] Ohio State Univ, Dept Evolut Ecol & Organismal Biol, Columbus, OH 43210 USA
[3] Ohio State Univ, Ohio Agr Res & Dev Ctr, Dept Entomol, Wooster, OH 44691 USA
[4] USDA ARS, Southern Plains Agr Res Ctr, Insect Control & Cotton Dis Res Unit, College Stn, TX 77845 USA
关键词
Diapause hormone; Receptor; Xenopus oocytes; Analogs; Diapause termination; PROTEIN-COUPLED RECEPTOR; HELIOTHIS-VIRESCENS; PROTHORACICOTROPIC HORMONE; PUPAL DIAPAUSE; TERMINATION; LEPIDOPTERA; PEPTIDES; AGONISTS; INDUCE; GLAND;
D O I
10.1016/j.peptides.2013.12.005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Diapause hormone (DH) is an insect neuropeptide that is highly effective in terminating the overwintering pupal diapause in members of the HelicoverpalHelio this complex of agricultural pests, thus DH and related compounds have promise as tools for pest management. To augment our development of effective DH analogs and antagonists that could be used as diapause disruptors this study focuses on the cloning and identification of the DH receptor (DHR) in the corn earworm, Helicoverpa zea. The full-length dhr cDNA contains 2153 nucleotides encoding 511 amino acids. Our results suggest there are at least two splicing variants of Hezea-DHR. Hydrophobicity analysis and sequence alignment indicate that Hezea-DHR has 7 transmembrane regions and a highly conserved C-terminal region that is also present in related receptors. Hezea-DHR has 95%, 82% and 79% identity to a partial DHR sequence from Heliothis virescens, a full-length DHR in Orgyia thyellina, and DHR-1 in Bombyx mori, but only 45-49% identity to pheromone biosynthesis activating neuropeptide receptor (PBANR). Expression of dhr mRNA remained low in whole body extracts throughout diapause and in young nondiapausing pupae, but was distinctly elevated as development ensued in pharate adults 7 days after pupation. The highest expression of dhr mRNA we noted was in the ovary. A DHR fusion protein with enhanced-green fluorescent protein was successfully expressed heterologously in X. laevis oocytes, as verified by fluorescent imaging and Western blots, but an electrophysiological assay failed to detect receptor-ligand binding activity, which suggests that an essential cofactor and/or accessory protein is required for functional activity of the DHR. (c) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码:250 / 257
页数:8
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