Yeast one-hybrid screening the potential regulator of CYP6B6 overexpression ofHelicoverpa armigeraunder 2-tridecanone stress

Abstract

In insect, the cytochrome P450 plays a pivotal role in detoxification to toxic allelochemicals. Helicoverpa armigera can tolerate and survive in 2-tridecanone treatment owing to the CYP6B6 responsive expression, which is controlled by some regulatory DNA sequences and transcription regulators. Therefore, the 2-tridecanone responsive region and transcription regulators of the CYP6B6 are responsible for detoxification of cotton bollworm. In this study, we used yeast one-hybrid to screen two potential transcription regulators of the CYP6B6 from H. armigera that respond to the plant secondary toxicant 2-tridecanone, which were named Prey1 and Prey2, respectively. According to the NCBI database blast, Prey1 is the homology with FK506 binding protein (FKBP) of Manduca sexta and Bombyx mori that belongs to the FKBP-C superfamily, while Prey2 may be a homology of an unknown protein of Papilio or the fcaL24 protein homology of B. mori. The electrophoretic mobility shift assays revealed that the FKBP of prokaryotic expression could specifically bind to the active region of the CYP6B6 promoter. After the 6th instar larvae of H. armigera reared on 2-tridecanone artificial diet, we found there were similar patterns of CYP6B6 and FKBP expression of the cotton bollworm treated with 10 mg g-1 2-tridecanone for 48 h, which correlation coefficient was the highest (0.923). Thus, the FKBP is identified as a strong candidate for regulation of the CYP6B6 expression, when the cotton bollworm is treated with 2-tridecanone. This may lead us to a better understanding of transcriptional mechanism of CYP6B6 and provide very useful information for the pest control.

Identification of the 2‐tridecanonecis‐acting element in the promoter of cytochrome P450CYP6B7inHelicoverpa armigera

“…In the present study, a cis ‐acting element responsible for 2‐tridecanone induction in the promoter of CYP6B7 was identified; however, it had no significant sequence similarity with the cis ‐acting elements reported previously, such as ARE, EcRE and XRE‐AhR, which have been well studied for their transcription factors. FK506 binding protein (FKBP) was screened as a candidate transcription factor for the regulation of CYP6B6 expression in H. armigera (Zhao et al ., ). As a homologous gene of CYP6B6 , whether the cis ‐acting element of CYP6B7 could be recognized by transcription factors of FKBP is unknown.…”

Alcohol dehydrogenase 5 of Helicoverpa armigera interacts with the CYP6B6 promoter in response to 2‐tridecanone

“…Competent cells of Escherichia coli strains DH5α and Transetta were purchased from TransGen Biotech. The yeast strains Y1HGold (p4r-AbAi) and Y1HGold (p4m-AbAi) were constructed by our research group as previously reported (Zhao et al, 2016b).…”

Insect Transcription Factors: A Landscape of Their Structures and Biological Functions in Drosophila and beyond

“…AhR heterodimerizes with aryl hydrocarbon receptor nuclear translocator (ARNT) to directly activate the expression of P450s after exposure to plant secondary metabolites [ 197 , 198 ]. In addition, two other potential TFs (FK506 binding protein (FKBP) and Prey2) were reported to regulate the expression of CYP6B6 in H. armigera under 2-tridecanone stress [ 199 ].…”