Functional characterization of Aedes aegypti alkaline phosphatase ALP1 involved in the toxicity of Cry toxins from Bacillus thuringiensis subsp. israelensis and jegathesan.
Functional characterization of Aedes aegypti alkaline phosphatase ALP1 involved in the toxicity of Cry toxins from Bacillus thuringiensis subsp. israelensis and jegathesan.
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DOI：
10.1016/j.peptides.2017.05.011
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发表时间：
2017-12
期刊：
影响因子：
3
通讯作者：
Gill SS
中科院分区：
文献类型：
--
作者：
Chen J;Aimanova K;Gill SS
目前，埃及伊蚊的三大类蛋白质，钙粘蛋白，碱性磷酸酶（ALP）和氨基肽酶N（APN），已被确定为Cry11Aa毒素受体。为了进一步表征它们对毒性的作用，先前产生了具有沉默的伊蚊钙粘蛋白表达的转基因蚊子，并且证明了钙粘蛋白在介导四种不同杀蚊毒素（Cry11Aa、Cry11Ba、Cry4Aa和Cry4Ba）的毒性中的作用。在这里，我们研究了另一种已报道的Cry11Aa受体ALP 1的作用。与伊蚊钙粘蛋白一样，这种蛋白质定位于远端和近端胃盲肠和后中肠的顶端细胞膜。我们还成功地产生了转基因蚊子，敲低ALP 1转录水平使用诱导型伊蚊热休克启动子，热休克70 A驱动dsALP 1RNA。使用四种不同的杀蚊毒素对这种转基因蚊子进行幼虫生物测定。生物测定结果表明，Cry11Aa对这些转基因幼虫的毒性在热休克后降低（4.4倍），Cry11Ba毒性略有减弱。但Cry4Aa和Cry4Ba对ALP 1沉默的幼虫的毒性没有变化。在没有热休克的情况下，所有四种毒素的毒性都没有改变，这表明该热休克启动子是热诱导的。值得注意的是，ALP1敲低的转基因蚊子对Cry11Aa毒素的抗性比伊蚊钙粘蛋白敲低的转基因蚊子低约3.7倍。这些结果表明，ALP 1是Cry11Aa和Cry11Ba的重要二级受体，但它可能不参与Cry4Aa和Cry4Ba毒性。
Presently three major groups of proteins from Aedes aegypti, cadherin, alkaline phosphatases (ALP) and aminopeptidases N (APN), have been identified as Cry11Aa toxin receptors. To further characterize their role on toxicity, transgenic mosquitoes with silenced Aedes cadherin expression were previously generated and the role of cadherin in mediating the toxicity of four different mosquitocidal toxins (Cry11Aa, Cry11Ba, Cry4Aa and Cry4Ba) was demonstrated. Here, we investigated the role of another reported Cry11Aa receptor, ALP1. As with Aedes cadherin, this protein is localized in the apical cell membrane of distal and proximal gastric caecae and the posterior midgut. We also successfully generated transgenic mosquitoes that knockdowned ALP1 transcript levels using an inducible Aedes heat shock promoter, Hsp70A driving dsALP1RNA. Four different mosquitocidal toxins were used for larval bioassays against this transgenic mosquito. Bioassay results show that Cry11Aa toxicity to these transgenic larvae following a heat shock decreased (4.4 fold) and Cry11Ba toxicity is slightly attenuated. But Cry4Aa and Cry4Ba toxicity to ALP1 silenced larvae is unchanged. Without heat shock, toxicity of all four toxins does not change, suggesting this heat shock promoter is heat-inducible. Notably, transgenic mosquitoes with ALP1 knockdown are about 3.7 times less resistant to Cry11Aa toxin than those with Aedes cadherin knockdown. These results demonstrate that the ALP1 is an important secondary receptor for Cry11Aa and Cry11Ba, but it might not be involved in Cry4Aa and Cry4Ba toxicity.
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影响因子：
3.7
作者：
Jurat-Fuentes JL;Karumbaiah L;Jakka SR;Ning C;Liu C;Wu K;Jackson J;Gould F;Blanco C;Portilla M;Perera O;Adang M
通讯作者：
Adang M
影响因子：
4.5
作者：
Gahan LJ;Pauchet Y;Vogel H;Heckel DG
通讯作者：
Heckel DG
影响因子：
2.4
作者：
Horn, C;Wimmer, EA
通讯作者：
Wimmer, EA
影响因子：
3
作者：
Devillers J;Lagneau C;Lattes A;Garrigues JC;Clémenté MM;Yébakima A
通讯作者：
Yébakima A
影响因子：
3.8
作者：
Lee SB;Aimanova KG;Gill SS
通讯作者：
Gill SS