Silencing of midgut aminopeptidase N of Spodoptera litura by double-stranded RNA establishes its role as bacillus thuringiensis toxin receptor
Silencing of midgut aminopeptidase N of Spodoptera litura by double-stranded RNA establishes its role as bacillus thuringiensis toxin receptor
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DOI：
10.1074/jbc.c200523200
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发表时间：
2002-12-06
影响因子：
4.8
通讯作者：
Bhatnagar, RK
中科院分区：
文献类型：
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作者：
Rajagopal, R;Sivakumar, S;Bhatnagar, RK
苏云金芽孢杆菌的杀虫晶体蛋白与敏感昆虫的中肠受体结合，导致昆虫的孔形成和死亡。受体的身份尚未明确确定。最近，通过免疫荧光和体外配体印迹相互作用证实了斜纹夜蛾（Spodoptera litura）克隆的异源表达的氨基肽酶（slapn）与Cry1C蛋白之间的直接相互作用。在这里，我们发现给斜纹夜蛾幼虫注射slapn双链RNA会降低其表达。由于表达量的减少，这些幼虫对Cry1C毒素的敏感性也相应降低。在昆虫的蜕皮和发育过程中，基因沉默被保留下来，并遗传给下一代，尽管效果有所降低。这些结果直接暗示幼虫中肠氨基肽酶N是苏云金芽孢杆菌杀虫蛋白的受体。
Insecticidal crystal proteins of Bacillus thuringiensis bind to receptors in the midgut of susceptible insects leading to pore formation and death of the insect. The identity of the receptor is not clearly established. Recently a direct interaction between a cloned and heterologously expressed aminopeptidase (slapn) from Spodoptera litura and the Cry1C protein was demonstrated by immunofluorescence and in vitro ligand blot interaction. Here we show that administration of slapn double-stranded RNA to S. litura larvae reduces its expression. As a consequence of the reduced expression, a corresponding decrease in the sensitivity of these larvae to Cry1C toxin was observed. The gene silencing was retained during the insect's moulting and development and transmitted to the subsequent generation albeit with a reduced effect. These results directly implicate larval midgut aminopeptidase N as receptor for Bacillus thuringiensis insecticidal proteins.