Functional analysis of ABCG and ABCH transporters from the red flour beetle, Tribolium castaneum.
Functional analysis of ABCG and ABCH transporters from the red flour beetle, Tribolium castaneum.
复制标题
DOI：
10.1002/ps.6332
复制
发表时间：
2021-02
影响因子：
4.1
通讯作者：
Janin Rösner;Johanne Tietmeyer;H. Merzendorfer
中科院分区：
文献类型：
--
作者：
Janin Rösner;Johanne Tietmeyer;H. Merzendorfer
背景ABCA-C和ABCG-H亚家族的ABC转运蛋白被认为与杀虫剂的解毒有关，主要是因为发现杀虫剂处理后基因表达水平升高。我们以前已经从模式甲虫和害虫卡斯滕姆中鉴定了TcABCA-C基因，并证明了特定的TcABCA和TcABCC基因参与了灭虫脲的消除，因为RNAi介导的基因沉默增加了敏感性。在本研究中，我们重点研究了TcABCG和TcABCH基因在杀虫剂解毒中的作用。结果当我们通过RNAi沉默TcABCG-H基因的表达时，我们注意到了一种以前未见报道的TcABCG-4F的发育RNAi表型，其特征是成体蜕皮时50%的死亡率和蜕皮停止。当我们击倒果蝇棕色同源基因TcABCG-XC时，我们没有获得明显的眼睛颜色表型，但观察到马氏管摄取核黄素的损失。接下来，我们测定了所有TcABCG-H基因在不同组织和发育阶段的表达谱，并分析了四种化学无关杀虫剂处理后的转录水平。事实上，我们发现一些基因在杀虫剂处理后特别上调。然而，当我们测定杀虫剂诱导的幼虫死亡率时，如果用dsRNA注射来沉默那些上调的TcABCG-H基因，我们没有观察到对杀虫剂的敏感性显著增加。结论杀虫剂依赖诱导TcABCG-H基因表达反映了一种非特异性的应激反应，从而强调了杀虫剂解毒功能研究的重要性。这篇文章受版权保护。版权所有。
BACKGROUND ABC transporters of subfamilies ABCA-C and ABCG-H have been suggested to be implicated in insecticide detoxification mostly based on the finding of elevated gene expression levels in response to insecticide treatment. We have previously characterized TcABCA-C genes from the model beetle and pest Tribolium castaneum and demonstrated that specific TcABCA and TcABCC genes are involved in the elimination of diflubenzuron, as RNAi-mediated gene silencing increased susceptibility. In this study, we focused on the potential functions of TcABCG and TcABCH genes in insecticide detoxification. RESULTS When we silenced the expression of TcABCG-H genes by RNAi, we noticed a previously unreported developmental RNAi phenotype for TcABCG-4F, which is characterized by a 50% mortality and ecdysial arrest during adult molt. When we knocked down the Drosophila brown orthologue TcABCG-XC, we did not obtain apparent eye color phenotypes, but observed a loss of riboflavin uptake by Malpighian tubules. Next, we determined the expression profiles of all TcABCG-H genes in different tissues and developmental stages, and analyzed transcript levels in response to the treatment with four chemically unrelated insecticides. Indeed, we found that some genes were particularly upregulated after insecticide treatment. However, when we determined insecticide-induced mortalities in larvae that where treated by dsRNA-injection to silence those TcABCG-H genes that were upregulated, we did not observe a significant increase in susceptibility to the insecticides. CONCLUSION Our findings suggest that the observed insecticide-dependent induction of TcABCG-H gene expression reflects an unspecific stress response, and hence underlines the significance of functional studies on insecticide detoxification. This article is protected by copyright. All rights reserved.