Identification of 20-hydroxyecdysone late-response genes in the chitin biosynthesis pathway.
Identification of 20-hydroxyecdysone late-response genes in the chitin biosynthesis pathway.
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甲壳质生物合成途径中 20-羟基蜕皮激素迟发反应基因的鉴定
DOI：
10.1371/journal.pone.0014058
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发表时间：
2010-11-18
期刊：
影响因子：
3.7
通讯作者：
Zhang W
中科院分区：
文献类型：
--
作者：
Yao Q;Zhang D;Tang B;Chen J;Chen J;Lu L;Zhang W
背景20-羟基蜕皮激素（20E）及其受体复合物蜕皮激素受体（EcR）和超气门（USP）在控制发育、变态、繁殖和滞育中发挥着至关重要的作用。配体-受体复合物 20E-EcR/USP 直接激活一小组早期反应基因和一大组晚期反应基因。然而，蜕皮激素反应基因之前尚未在昆虫几丁质生物合成的背景下进行表征。主要发现在这里，我们表明，针对鳞翅目昆虫甜菜夜蛾中两种 SeEcR 亚型的共同区域的基于注射的 RNA 干扰 (RNAi) 是有效的，其表型包括化蛹前的高死亡率和发育缺陷。基因特异性RNAi后，异常幼虫角质层中的几丁质含量显着下降。注射EcR dsRNA 72小时前，几丁质生物合成途径中的五个基因SeTre-1、SeG6PI、SeUAP、SeCHSA和SeCHSB的表达水平显着降低，而SeTre-2的表达在注射EcR dsRNA后72小时之前没有差异。同时，体内注射20E诱导上述5个基因的表达。此外，SeTre-1、SeG6PI、SeUAP和SeCHSB基因对激素表现出晚期反应，并且SeTre-1、SeG6PI、SeUAP和SeCHSB基因在20E时的诱导能够在体外被蛋白质合成抑制剂放线菌酮抑制，表明这些基因是20E晚期反应基因。结论 甲壳素生物合成途径中的SeTre-1、SeG6PI、SeUAP和SeCHSB为20E迟发型反应基因，20E及其特异性受体通过诱导其表达，在甲壳素生物合成调控中发挥关键作用。
Background 20-hydroxyecdysone (20E) and its receptor complex ecdysone receptor (EcR) and ultraspiracle (USP) play a crucial role in controlling development, metamorphosis, reproduction and diapause. The ligand-receptor complex 20E-EcR/USP directly activates a small set of early-response genes and a much larger set of late-response genes. However, ecdysone-responsive genes have not been previously characterized in the context of insect chitin biosynthesis. Principal Findings Here, we show that injection-based RNA interference (RNAi) directed towards a common region of the two isoforms of SeEcR in a lepidopteron insect Spodoptera exigua was effective, with phenotypes including a high mortality prior to pupation and developmental defects. After gene specific RNAi, chitin contents in the cuticle of an abnormal larva significantly decreased. The expression levels of five genes in the chitin biosynthesis pathway, SeTre-1, SeG6PI, SeUAP, SeCHSA and SeCHSB, were significantly reduced, while there was no difference in the expression of SeTre-2 prior to 72 hr after injection of EcR dsRNA. Meanwhile, injection of 20E in vivo induced the expression of the five genes mentioned above. Moreover, the SeTre-1, SeG6PI, SeUAP and SeCHSB genes showed late responses to the hormone and the induction of SeTre-1, SeG6PI, SeUAP and SeCHSB genes by 20E were able to be inhibited by the protein synthesis inhibitor cycloheximide in vitro indicating these genes are 20E late-response genes. Conclusions We conclude that SeTre-1, SeG6PI, SeUAP and SeCHSB in the chitin biosynthesis pathway are 20E late-response genes and 20E and its specific receptors plays a key role in the regulation of chitin biosynthesis via inducing their expression.
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影响因子：
2.7
作者：
ASHBURNER, M;RICHARDS, G
通讯作者：
RICHARDS, G
影响因子：
2.7
作者：
ASHBURNER, M
通讯作者：
ASHBURNER, M
影响因子：
64.5
作者：
KOELLE, MR;TALBOT, WS;HOGNESS, DS
通讯作者：
HOGNESS, DS
影响因子：
2.6
作者：
Basso, LR;Vasconcelos, C;Paçó-Larson, ML
通讯作者：
Paçó-Larson, ML
影响因子：
3.8
作者：
Chen, Xiaofei;Yang, Xing;Zhang, Wenqing
通讯作者：
Zhang, Wenqing