Transcription-inducing activity of natural and synthetic juvenile hormone agonists through the Drosophila Methoprene-tolerant protein.

Drosophila melanogaster Methoprene-tolerant Taiman juvenile hormone agonists luciferase reporter assay

Journal

Pest management science
ISSN: 1526-4998
Titre abrégé: Pest Manag Sci
Pays: England
ID NLM: 100898744

Informations de publication

Date de publication:
Jul 2020
Historique:
received: 14 11 2019
revised: 15 01 2020
accepted: 30 01 2020
pubmed: 1 2 2020
medline: 7 8 2020
entrez: 1 2 2020
Statut: ppublish

Résumé

Juvenile hormones (JHs) are a class of sesquiterpenoids that play a pivotal role in insect growth and reproduction. Synthetic JH agonists (JHAs), including pyriproxyfen, have been widely used as insecticides to control agricultural pests and disease vectors. The antimetamorphic action of JHAs is mediated by their intracellular receptor, the heterodimer of Methoprene-tolerant (Met) and Taiman (Tai) proteins. Although a range of bioassay systems has been developed to detect the activity of JHAs, each of these systems has its own drawback(s), such as poor reproducibility, the use of radioactive ligands or the effect of endogenous JH-signaling factors. To address these issues, we constructed a new luciferase reporter assay for JHAs in mammalian HEK293T cells transiently transfected with the Drosophila Met and Tai genes. This reporter system gave highly reproducible results and showed nanomolar sensitivity to natural JHs. We then applied this reporter system to a structure-activity relationship (SAR) analysis of 14 natural and synthetic JHAs, leading to identification of the ligand structural factors important for the transcription-inducing activity. Because this reporter system is not affected by the signaling cascade downstream of the JH receptors, it is suitable for evaluating the intrinsic activity of JHAs. The SAR results obtained in this study therefore provide invaluable information on the rational design of novel JHA insecticides.

Sections du résumé

BACKGROUND BACKGROUND
Juvenile hormones (JHs) are a class of sesquiterpenoids that play a pivotal role in insect growth and reproduction. Synthetic JH agonists (JHAs), including pyriproxyfen, have been widely used as insecticides to control agricultural pests and disease vectors. The antimetamorphic action of JHAs is mediated by their intracellular receptor, the heterodimer of Methoprene-tolerant (Met) and Taiman (Tai) proteins. Although a range of bioassay systems has been developed to detect the activity of JHAs, each of these systems has its own drawback(s), such as poor reproducibility, the use of radioactive ligands or the effect of endogenous JH-signaling factors.
RESULTS RESULTS
To address these issues, we constructed a new luciferase reporter assay for JHAs in mammalian HEK293T cells transiently transfected with the Drosophila Met and Tai genes. This reporter system gave highly reproducible results and showed nanomolar sensitivity to natural JHs. We then applied this reporter system to a structure-activity relationship (SAR) analysis of 14 natural and synthetic JHAs, leading to identification of the ligand structural factors important for the transcription-inducing activity.
CONCLUSION CONCLUSIONS
Because this reporter system is not affected by the signaling cascade downstream of the JH receptors, it is suitable for evaluating the intrinsic activity of JHAs. The SAR results obtained in this study therefore provide invaluable information on the rational design of novel JHA insecticides.

Identifiants

pubmed: 32003111
doi: 10.1002/ps.5766
doi:

Substances chimiques

Drosophila Proteins 0
Methoprene 8B830OJ2UX

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

2316-2323

Subventions

Organisme : Japan Society for the Promotion of Science
ID : JP16K07625
Organisme : Japan Society for the Promotion of Science
ID : JP17J01486
Organisme : Japan Society for the Promotion of Science
ID : JP19K06051

Informations de copyright

© 2020 Society of Chemical Industry.

Références

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Auteurs

Taiyo Yokoi (T)

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

Taku Nabe (T)

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

Chiharu Ishizuka (C)

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

Ken'ichiro Hayashi (K)

Department of RNA Biology and Neuroscience, Graduate School of Medicine, Osaka University, Suita, Japan.

Sayoko Ito-Harashima (S)

Department of Biology, Graduate School of Science, Osaka Prefecture University, Sakai, Japan.

Takashi Yagi (T)

Department of Biology, Graduate School of Science, Osaka Prefecture University, Sakai, Japan.

Yoshiaki Nakagawa (Y)

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

Hisashi Miyagawa (H)

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

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Classifications MeSH