Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
Insect Biochem Mol Biol. 2010 Jan;40(1):58-63. doi: 10.1016/j.ibmb.2009.12.010. Epub 2009 Dec 28.
The insecticidal Cry toxins are pore-forming toxins produced by the bacteria Bacillus thuringiensis that disrupt insect-midgut cells. In this work we analyzed the response of two different insect orders, the Lepidopteran Manduca sexta and Dipteran Aedes aegypti to highly specific Cry toxins, Cry1Ab and Cry11Aa, respectively. One pathway activated in different organisms in response to a variety of pore-forming toxins is the mitogen-activated protein kinase p38 pathway (MAPK p38) that activates a complex defense response. We analyzed the MAPK p38 activation by immunodetection of its phosphorylated isoform, and the induction of p38 by RT-PCR, real-time PCR quantitative assays and immunodetection. We show that MAPK p38 is activated at postraductional level after Cry toxin intoxication in both insect orders. We detected the p38 induction at the transcriptional and traductional level, and observed a different response. In these three levels, we found that both insects respond to Cry toxin action but M. sexta responses more strongly than A. aegypti. Gene silencing of MAPK p38 in vivo, resulted in both insect species becoming hypersensitive to Cry toxin action, suggesting that the MAPK p38 pathway is involved in insect defense against Bt Cry toxins. This finding may have biotechnological applications for enhancing the activity of some Bt Cry toxins against specific insect pests.
杀虫晶体蛋白 Cry 毒素是由苏云金芽孢杆菌产生的一种形成孔的毒素,它破坏昆虫中肠细胞。在这项工作中,我们分析了两个不同昆虫目,鳞翅目 Manduca sexta 和双翅目 Aedes aegypti 对高度特异性 Cry 毒素 Cry1Ab 和 Cry11Aa 的反应。一种在不同生物体中响应各种形成孔毒素的途径是丝裂原活化蛋白激酶 p38 途径(MAPK p38),它激活了复杂的防御反应。我们通过免疫检测其磷酸化同工型来分析 MAPK p38 的激活,并通过 RT-PCR、实时 PCR 定量测定和免疫检测分析 p38 的诱导。我们表明,MAPK p38 在两种昆虫目中毒素后翻译后水平被激活。我们在转录和翻译水平检测到 p38 的诱导,并观察到不同的反应。在这三个水平上,我们发现两种昆虫都对 Cry 毒素的作用有反应,但 M. sexta 的反应比 A. aegypti 更强。体内 MAPK p38 的基因沉默导致两种昆虫对 Cry 毒素的作用变得更加敏感,这表明 MAPK p38 途径参与了昆虫对 Bt Cry 毒素的防御。这一发现可能具有生物技术应用,可增强某些 Bt Cry 毒素对特定害虫的活性。