Binding of de novo synthesized radiolabeled juvenile hormone (JH III) by JH receptors from the Cuban subterranean termite Prorhinotermes simplex and the German cockroach Blattella germanica

Miláček, Matěj; Bittova, Lenka; Tumova, Sarka; Lukšan, Ondřej; Hanus, Robert; Kyjaková, Pavlína; Machara, Aleš; Marek, Aleš; Jindra, Marek

doi:10.1016/j.ibmb.2021.103671

Cited by 9 publications

(10 citation statements)

References 60 publications

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“…JH agonists, such as methoprene and pyriproxyfen, were found to bind to germ cell expressed ( Gce ), a paralog of Met in Drosophila melanogaster 23,24 . However, the JH binding assay requires tritium‐labeled JH III ([ 3 H]‐JH III), which is currently unavailable for purchase and is difficult to synthesize because of the need for specialized equipment and synthesis facilities 25 . Therefore, we evaluated the JH antagonistic activity more conveniently in a luciferase reporter assay instead of the JH binding assay 15 .…”

Section: Resultsmentioning

confidence: 99%

“…23,24 However, the JH binding assay requires tritium-labeled JH III ([ 3 H]-JH III), which is currently unavailable for purchase and is difficult to synthesize because of the need for specialized equipment and synthesis facilities. 25 Therefore, we evaluated the JH antagonistic activity more conveniently in a luciferase reporter assay instead of the JH binding assay. 15 Figure 3 shows that both NY04 and EMBP inhibited the expression of reporter gene induced by JH I in a concentration-dependent manner.…”

Section: Jh Signaling Inhibitory Activity Of Embpmentioning

confidence: 99%

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Synthesis of 1,4‐benzodioxan derivatives and the evaluation of their biological activity as a novel juvenile hormone signaling inhibitor

Furuta,

Yamada,

Kayukawa

2023

Pest Management Science

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BACKGROUNDJuvenile hormone (JH) signaling inhibitors may be used as insect growth regulators due to their ability to control metamorphosis and reproduction in insects by regulating the action of JH.RESULTSWe identified ethyl (E)‐3‐(4‐{[7‐ (4‐methoxycarbonylbenzyloxy)‐1,4‐benzodioxan‐6‐yl]methyl}phenyl)prop‐2‐enoate (EMBP) and observed its strong precocious metamorphosis‐inducing activity against silkworm larvae. To further elucidate its mechanism of action, we investigated the effect of EMBP on JH‐mediated signaling pathway in vitro and in vivo. In a reporter assay using a Bombyx mori cell line, EMBP strongly suppressed the induction of reporter gene expression by JH I in a concentration‐dependent manner. A parallel rightward shift was observed in the dose–response curve of JH I after treatment with EMBP, indicating that EMBP competitively inhibited JH. Moreover, we monitored the developmental changes in the JH‐responsive gene, Krüppel homolog 1 (Kr‐h1), and ecdysone‐responsive gene, Broad‐Complex (BRC), in EMBP‐treated silkworm larvae. EMBP suppressed only the expression of Kr‐h1 in third instar larvae.CONCLUSIONOur results demonstrated that EMBP specifically regulates the JH‐mediated Kr‐h1 signaling pathway. EMBP could be used as a lead compound in the development of new insect growth regulators.This article is protected by copyright. All rights reserved.

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Section: Resultsmentioning

confidence: 99%

Section: Jh Signaling Inhibitory Activity Of Embpmentioning

confidence: 99%

Synthesis of 1,4‐benzodioxan derivatives and the evaluation of their biological activity as a novel juvenile hormone signaling inhibitor

Furuta,

Yamada,

Kayukawa

2023

Pest Management Science

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“…Additional reporter plasmids, JHRE [nLuc] and mut JHRE [nLuc], were prepared by substituting the firefly luciferase sequence with that encoding the NanoLuc® luciferase (Promega, Madison, WI, USA). The NanoLuc® sequence was excised from a previously described vector pNL-9xUAS [nLucP Hygro] 62 using Hin dIII and Bam HI, and ligated into JHRE [fLuc] and mut JHRE [fLuc] using the same restriction enzymes. EGFP sequence cloned into the pIEx-4 plasmid (Sigma-Aldrich, Saint Louis, MO, USA) served as a negative control for cell line selection.…”

Section: Methodsmentioning

confidence: 99%

Unique and Common Agonists Activate the Insect Juvenile Hormone Receptor and the Human AHR

Sedlak,

Tuma,

Kolla

et al. 2024

Preprint

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Transcription factors of the bHLH-PAS family play vital roles in animal development, physiology, and disease. Two members of the family require binding of low-molecular weight ligands for their activity: the vertebrate aryl hydrocarbon receptor (AHR) and the insect juvenile hormone receptor (JHR). In the fly Drosophila melanogaster, the paralogous proteins GCE and MET constitute the ligand-binding component of JHR complexes. Whilst GCE/MET and AHR are phylogenetically heterologous, their mode of action is similar. JHR is targeted by several synthetic agonists that serve as insecticides disrupting the insect endocrine system. AHR is an important regulator of human endocrine homeostasis and it responds to environmental pollutants and endocrine disruptors. Whether AHR signaling is affected by compounds that can activate JHR has not been reported. To address this question, we screened a chemical library of 50,000 compounds to identify 93 novel JHR agonists in a reporter system based on Drosophila cells. Of these compounds, 26% modulated AHR signaling in an analogous reporter assay in a human cell line, indicating a significant overlap in the agonist repertoires of the two receptors. To explore the structural features of agonist-dependent activation of JHR and AHR, we compared the ligand-binding cavities and their interactions with selective and common ligands of AHR and GCE. Molecular dynamics modeling revealed ligand-specific as well as conserved side chains within the respective cavities. Significance of predicted interactions was supported through site-directed mutagenesis. The results have indicated that synthetic insect juvenile hormone agonists might interfere with AHR signaling in human cells.

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“…Synthetic DNA sequence encoding amino acids M1‐V505 of T. castaneum Tai (same as in the NanoBiT constructs above) was cloned to a pDBD vector based on the pcDNA3.1/Zeo(+) plasmid (ThermoFisher Scientific) with an insert of DNA encoding amino acids M1‐S147 of the Gal4 DNA‐binding domain (DBD). A reporter construct pNL‐9xUAS [16] carried nine repeats of the upstream activation sequence ( UAS ) before a minimal promoter in the pNL[NLucP/minP/Hygro] vector (Promega) encoding NanoLuc luciferase fused to the PEST destabilisation sequence (NLucP) (Promega). The pACT[VP16‐Met] and pDBD[Gal4‐Tai] constructs (each 27 ng DNA per well) were co‐transfected along with 110 ng per well of the pNL‐9xUAS reporter plasmid to semiconfluent CHO cells seeded in 48‐well plates.…”

Section: Methodsmentioning

confidence: 99%

“…JHs comprise a group of insect-specific, epoxidated and methylated sesquiterpenoids produced by the corpora allata glands [9,10]. Intracellular receptors that bind JHs with high affinity have been identified as protein products of the Methoprene-tolerant (Met) gene [11] and its orthologs in the flour beetle Tribolium castaneum [12], the fruit fly Drosophila melanogaster [13,14] and other insects [15,16]. Unlike nuclear receptors for other lipophilic hormones, the JH receptor (JHR) Met belongs to the bHLH-PAS family of transcription factors [17].…”

Section: Introductionmentioning

confidence: 99%

Ligand‐dependent protein interactions of the juvenile hormone receptor captured in real time

Tumova

Jindra

2023

The FEBS Journal

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Juvenile hormone (JH) signalling provides vital regulatory functions during insect development via transcriptional regulation of genes critical for the progression of metamorphosis and oogenesis. Despite the importance of JH signalling, the underlying molecular mechanisms remain largely unknown. Our current understanding of the pathway depends on static end-point information and suffers from the lack of time-resolved data.Here, we have addressed the dynamic aspect of JH signalling by monitoring in real time the interactions of insect JH receptor proteins. Use of two tags that reconstitute a functional luciferase when in proximity enabled us to follow the rapid assembly of a JH receptor heterodimer from basic helix-loop-helix/Per-Arnt-SIM (bHLH-PAS) proteins, methoprene-tolerant (Met) and taiman (Tai), upon specific JH binding to Met. On a similar timescale (minutes), the dissociation of Met-Met complexes occurred, again strictly dependent on Met interaction with specific agonist ligands. To resolve questions regarding the regulatory role of the chaperone Hsp90/83 in the JHR complex formation, we used the same technique to demonstrate that the Met-Hsp83 complex persisted in the agonist absence but readily dissociated upon specific binding of JH to Met. Preincubation with the Hsp90 inhibitor geldanamycin showed that the chaperone interaction protected Met from degradation and was critical for Met to produce the active signalling dimer with Tai. Thus, the JH receptor functions appear to be governed by principles similar to those regulating the aryl hydrocarbon receptor, the closest vertebrate homologue of the arthropod JH receptor.

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Binding of de novo synthesized radiolabeled juvenile hormone (JH III) by JH receptors from the Cuban subterranean termite Prorhinotermes simplex and the German cockroach Blattella germanica

Cited by 9 publications

References 60 publications

Synthesis of 1,4‐benzodioxan derivatives and the evaluation of their biological activity as a novel juvenile hormone signaling inhibitor

Synthesis of 1,4‐benzodioxan derivatives and the evaluation of their biological activity as a novel juvenile hormone signaling inhibitor

Unique and Common Agonists Activate the Insect Juvenile Hormone Receptor and the Human AHR

Ligand‐dependent protein interactions of the juvenile hormone receptor captured in real time

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