Nakagawa Yoshiaki, Sakai Atsushi, Magata Fumie, Ogura Takehiko, Miyashita Masahiro, Miyagawa Hisashi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
FEBS J. 2007 Dec;274(23):6191-203. doi: 10.1111/j.1742-4658.2007.06139.x.
cDNAs of the ecdysone receptor and the retinoid X receptor were cloned from the Japanese scorpion Liocheles australasiae, and the amino acid sequences were deduced. The full-length cDNA sequences of the L. australasiae ecdysone receptor and the L. australasiae retinoid X receptor were 2881 and 1977 bp in length, respectively, and the open reading frames encoded proteins of 560 and 414 amino acids. The amino acid sequence of the L. australasiae ecdysone receptor was similar to that of the ecdysone receptor-A of the soft tick, Ornithodoros moubata (68%) and to that of the ecdysone receptor-A1 of the lone star tick, Amblyomma americanum (66%), but showed lower similarity to the ecdysone receptors of Orthoptera and Coleoptera (53-57%). The primary sequence of the ligand-binding region of the L. australasiae ecdysone receptor was highly homologous to that of ticks (85-86%). The amino acid sequence of the L. australasiae retinoid X receptor was also homologous to the amino acid sequence of ultraspiracles of ticks (63%) and insects belonging to the orders Orthoptera and Coleoptera (60-64%). The identity of both the L. australasiae ecdysone receptor and the L. australasiae retinoid X receptor to their lepidopteran and dipteran orthologs was less than 50%. The cDNAs of both the L. australasiae ecdysone receptor (L. australasiae ecdysone receptor-A) and the L. australasiae retinoid X receptor were successfully translated in vitro using a rabbit reticulocyte lysate system. An ecdysone analog, ponasterone A, bound to L. australasiae ecdysone receptor-A (K(D) = 4.2 nM), but not to L. australasiae retinoid X receptor. The L. australasiae retinoid X receptor did not enhance the binding of ponasterone A to L. australasiae ecdysone receptor-A, although L. australasiae retinoid X receptor was necessary for the binding of L. australasiae ecdysone receptor-A to ecdysone response elements.
从日本蝎子澳链尾蝎中克隆了蜕皮激素受体和维甲酸X受体的cDNA，并推导了其氨基酸序列。澳链尾蝎蜕皮激素受体和澳链尾蝎维甲酸X受体的全长cDNA序列分别为2881和1977 bp，开放阅读框编码的蛋白质分别为560和414个氨基酸。澳链尾蝎蜕皮激素受体的氨基酸序列与软蜱钝缘蜱的蜕皮激素受体-A（68%）以及美洲钝眼蜱的蜕皮激素受体-A1（66%）相似，但与直翅目和鞘翅目的蜕皮激素受体的相似性较低（53 - 57%）。澳链尾蝎蜕皮激素受体配体结合区的一级序列与蜱的高度同源（85 - 86%）。澳链尾蝎维甲酸X受体的氨基酸序列也与蜱的超气门蛋白（63%）以及直翅目和鞘翅目昆虫的氨基酸序列同源（60 - 64%）。澳链尾蝎蜕皮激素受体和澳链尾蝎维甲酸X受体与它们鳞翅目和双翅目直系同源物的一致性均小于50%。使用兔网织红细胞裂解物系统在体外成功翻译了澳链尾蝎蜕皮激素受体（澳链尾蝎蜕皮激素受体-A）和澳链尾蝎维甲酸X受体的cDNA。一种蜕皮激素类似物，蜕皮甾酮A，与澳链尾蝎蜕皮激素受体-A结合（K(D)=4.2 nM），但不与澳链尾蝎维甲酸X受体结合。尽管澳链尾蝎维甲酸X受体是澳链尾蝎蜕皮激素受体-A与蜕皮激素反应元件结合所必需的，但它并未增强蜕皮甾酮A与澳链尾蝎蜕皮激素受体-A的结合。
