Molecular characterization of methoprene-tolerant gene (Met) in the swimming crab Portunus trituberculatus: Its putative role in methyl farnesoate-mediated vitellogenin transcriptional activation
Molecular characterization of methoprene-tolerant gene (Met) in the swimming crab Portunus trituberculatus: Its putative role in methyl farnesoate-mediated vitellogenin transcriptional activation
复制标题
三疣梭子蟹中甲虫烯耐受基因（Met）的分子特征：其在法呢酸甲酯介导的卵黄蛋白原转录激活中的假定作用
DOI：
10.1016/j.anireprosci.2016.09.017
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发表时间：
2016-11-01
影响因子：
2.2
通讯作者：
Zhu, Dongfa
中科院分区：
文献类型：
--
作者：
Liu, Mingxin;Xie, Xi;Zhu, Dongfa
耐甲氧普仑（Methoprene-tolerant，Met）属于核转录调节因子的碱性螺旋-环-螺旋（basic helix-loop-helix，bHLH）-Per-Arnt-Sim（per-Arnt-Sim，PAS）家族，并且是昆虫保幼激素（Juvenile hormone，OH）和甲壳类动物法呢酸甲酯（methyl farnesoate，MF）的主要候选受体。本研究从三疣梭子蟹（Portunus tritubercututus，PtMet）中克隆了Met的全长cDNA。发现PtMet氨基酸序列含有bHLH-PAS家族蛋白的特征性结构域，并且具有几个特异性负责JH或MF结合的保守氨基酸残基。除了其广泛的组织表达，PtMet mRNA在雌性肝胰腺中高度表达，肝胰腺是三疣梭子蟹卵黄蛋白原（Vg）合成的主要部位。PtMet在卵巢发育的卵黄发生早期表达显著增加，可能参与卵黄发生的调控。在体外，MF诱导上调PtMet和PtVg的表达在肝胰腺，但不是在卵巢。在体内，MF诱导PtMet和PtVg在肝胰腺中的表达，并且在卵巢中仅诱导PtMet的表达。综上所述，这些结果表明，Met可以潜在地作为甲壳类动物中的MF受体。此外，Met与MF的结合可能在转录水平上调节肝胰腺中Vg的合成。(C)2016爱思唯尔B. V.保留所有权利。
Methoprene-tolerant (Met) belongs to the basic helix-loop-helix (bHLH)-Per-Arnt-Sim (PAS) family of nuclear transcriptional regulators, and is the leading candidate receptor for the insect Juvenile hormone OH) and the crustacean methyl farnesoate (MF). In the present study, a full-length cDNA of Met was cloned from the swimming crab, Portunus tritubercututus (PtMet). The PtMet amino acid sequence was found to contain domains characteristic of the bHLH-PAS family proteins and to have several conserved amino acid residues specifically responsible for JH or MF binding. In addition to its extensive tissue expression, PtMet mRNA is highly expressed in the female hepatopancreas, the primary site of vitellogenin (Vg) synthesis in P. trituberculatus. PtMet expression was significantly increased in the early-vitellogenic stage of ovarian development, which indicated that likely involvement in vitellogenesis regulation. In vitro, MF induced up-regulation of PtMet and PtVg expression in the hepatopancreas, but not in the ovary. In vivo, MF induced PtMet and PtVg expression in the hepatopancreas,and only PtMet expression was induced in the ovary. Taken together, these results suggest that Met can potentially serve as a MF receptor in crustaceans. In addition, the binding of Met to MF might transcriptionally regulate Vg synthesis in the hepatopancreas. (C) 2016 Elsevier B.V. All rights reserved.