Molecular cloning and mRNA expression of a ryanodine receptor gene in the cotton bollworm, Helicoverpa armigera
Molecular cloning and mRNA expression of a ryanodine receptor gene in the cotton bollworm, Helicoverpa armigera
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棉铃虫兰尼碱受体基因的分子克隆及 mRNA 表达
DOI：
10.1016/j.pestbp.2013.09.006
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发表时间：
2013-11-01
影响因子：
4.7
通讯作者：
Wang, Jianjun
中科院分区：
文献类型：
--
作者：
Wang, Jian;Liu, Yaping;Wang, Jianjun
Ryanodine受体（RyRs）是新型双酰胺类杀虫剂的靶标。棉铃虫Helicoverpa armigera是世界上最重要的棉花害虫之一。本研究报道了黑腹果蝇RyR基因全长cDNA序列（命名为HaRyR）。armigera。该序列全长16，083 bp，编码5，142个氨基酸，与褐飞虱（Nilaparvatalugens，NlRyR）和黑腹果蝇（Drosophilamelanogaster，DmRyR）同源性分别为80%和78%。RyR蛋白的所有特征在HaRyR中是保守的，包括在Ca 2+释放通道中保守的GXRXGGGXGD基序和RyR通道特有的RyR结构域的四个拷贝。在HaRyR中也发现了先前鉴定的七个鳞翅目特异性RyR残基（N-4977、N-4979、N-4990、L-5005、L-5036、N-5068和T-5119）。氨基酸序列比对显示HaRyR的N-末端区域（残基188-295）与NIRyR（94%）和DmRyR（92%）共享高序列同一性，并且与三种兔RyR同种型共享中等序列同一性（47-50%），而HaRyR的C末端跨膜区的短片段（残基4632-4676）显示与NlRyR（69%）和DmRyR（67%）的中等序列同一性，以及与三种兔RyR同种型的低序列同一性（19-28%）。HaRyR基因在卵中的表达量显著低于3龄幼虫、蛹和成虫，HaRyR基因在卵中的表达存在解剖学规律，头部的表达量最高，而胸部和腹部的表达量最低。本研究结果为HaRyR的结构和功能表征以及深入了解双酰胺类杀虫剂在不同物种间毒性选择性的分子机制奠定了基础。(C)2013 Elsevier Inc. All rights reserved.
Ryanodine receptors (RyRs) are the targets of novel diamide insecticides. The cotton bollworm, Helicoverpa armigera, is one of the most important cotton pests in the world. In this study, we report the full-length RyR cDNA sequence (named as HaRyR) of H. armigera. The 16,083-bp contiguous sequence encoded 5, 142 amino acid residues, which shares 80% and 78% overall identities with its homologues in Nilaparvata lugens (NlRyR) and Drosophila melanogaster (DmRyR), respectively. All hallmarks of RyR proteins are conserved in the HaRyR, including the GXRXGGGXGD motif conserved in the Ca2+ release channels and four copies of RyR domain unique to RyR channels. The previously identified seven lepidopteran-specific RyR residues were also found in HaRyR (N-4977, N-4979, N-4990, L-5005, L-5036, N-5068 and T-5119). An amino acid sequence alignment showed that the N-terminal region of HaRyR (residues 188-295) shared high sequence identity with NIRyR (94%) and DmRyR (92%), and moderate sequence identity (47-50%) with three rabbit RyR isoforms, while the short segment of the C-terminal transmembrane region of HaRyR (residues 4632-4676) exhibited moderate sequence identity with NlRyR (69%) and DmRyR (67%), and low sequence identity (19-28%) with three rabbit RyR isoforms. In addition, expression analysis of HaRyR revealed that the mRNA expression level in eggs was significantly lower than in third instar larvae, pupae and adults, and anatomical regulation of HaRyR expression was also observed with the highest expression level in head compared with thorax and abdomen. Our results lay a foundation for comprehensive structural and functional characterization of HaRyR and for understanding of the molecular mechanisms of toxicity selectivity of diamide insecticides among different species. (C) 2013 Elsevier Inc. All rights reserved.