Innovation Center of Pesticide Research, Department of Applied Chemistry, College of Science, China Agricultural University, Beijing 100193, China.
College of Resources and Environment and Henan Engineering Research Center of Green Pesticide Creation &Intelligent Pesticide Residue Sensor Detection, Henan Institute of Science and Technology, Xinxiang 453003, Henan Province, China.
Int J Biol Macromol. 2024 Feb;258(Pt 2):128995. doi: 10.1016/j.ijbiomac.2023.128995. Epub 2023 Dec 28.
As we known, inducibility is an important feature of P450 genes. Previous studies indicated that CYP6B7 could be induced and involved in fenvalerate detoxification in Helicoverpa armigera. However, the regulatory mechanism of CYP6B7 induced by fenvalerate is still unclear. In this study, CYP6B7 promoter of H. armigera was cloned and the cis-acting element of fenvalerate was identified to be located between -84 and - 55 bp of CYP6B7 promoter. Subsequently, 33 candidate transcription factors (CYP6B7-fenvalerate association proteins, CAPs) that may bind to the cis-acting element were screened and verified by yeast one-hybrid. Among them, the expression levels of several CAPs were significantly induced by fenvalerate. Knockdown of juvenile hormone-binding protein-like (JHBP), RNA polymerase II-associated protein 3 (RPAP3), fatty acid synthase-like (FAS) and peptidoglycan recognition protein LB-like (PGRP) resulted in significant expression inhibition of CYP6B7, and increased sensitivity of H. armigera to fenvalerate. Co-transfection of reporter gene p (-84/-55) with pFast-CAP showed that JHBP, RPAP3 and PGRP could significantly increase the activity of CYP6B7 promoter. These results suggested that trans-acting factors JHBP, RPAP3 and PGRP may bind with cis-acting elements to regulate the expression of CYP6B7 induced by fenvalerate, and play an important role in the detoxification of H. armigera to fenvalerate.
众所周知，可诱导性是 P450 基因的一个重要特征。先前的研究表明，CYP6B7 可以被诱导，并参与棉铃虫对氰戊菊酯的解毒作用。然而，氰戊菊酯诱导 CYP6B7 的调控机制尚不清楚。本研究克隆了棉铃虫 CYP6B7 启动子，鉴定出 CYP6B7 启动子中 -84 至-55bp 之间的顺式作用元件是氰戊菊酯的作用位点。随后，通过酵母单杂交筛选并验证了 33 个可能与顺式作用元件结合的候选转录因子（CYP6B7-氰戊菊酯关联蛋白，CAPs）。其中，几种 CAPs 的表达水平被氰戊菊酯显著诱导。RNA 聚合酶 II 相关蛋白 3（RPAP3）、脂肪酸合酶样（FAS）和肽聚糖识别蛋白 LB 样（PGRP）的基因被敲低后，CYP6B7 的表达显著受到抑制，棉铃虫对氰戊菊酯的敏感性也显著增加。报告基因 p（-84/-55）与 pFast-CAP 共转染后发现，JHBP、RPAP3 和 PGRP 可以显著增加 CYP6B7 启动子的活性。这些结果表明，反式作用因子 JHBP、RPAP3 和 PGRP 可能与顺式作用元件结合，调节 CYP6B7 对氰戊菊酯的诱导表达，在棉铃虫对氰戊菊酯的解毒中发挥重要作用。
