Ecdysone Receptors of Pest Insects–Molecular Cloning, Characterisation, and a Ligand Binding Domain-Based Fluorescence Polarization Screen

Graham, Lloyd D.; Johnson, Wynona M.; Tohidi-Esfahani, Donya; Pawlak-Skrzecz, Anna; Bliese, Marianne; Lovrecz, George O.; Lu, Lu; Howell, Linda; Hannan, Garry N.; Hill, Ronald J.

doi:10.1007/978-1-4020-9112-4_19

Cited by 6 publications

(16 citation statements)

References 72 publications

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“…This finding is probably due to the fact that the structures of the EcR ligand-binding domains required for tebufenozide binding are very different among insect of different orders and arthropods. It is also known that the binding affinity of tebufenozide is negatively correlated with the distance in the phylogenetic trees constructed for the EcR ligand-binding domains (Graham et al, 2009). The sequence similarity of the EcR ligand-binding domains between L. australasiae and C. suppressalis is only 54%, suggesting that the EcR of L. australasiae cannot accommodate tebufenozide in its ligand-binding pocket.…”

Section: Binding Affinity Of Ecdysteroids To Ecrsmentioning

confidence: 97%

LC/MS/MS identification of 20-hydroxyecdysone in a scorpion (Liocheles australasiae) and its binding affinity to in vitro-translated molting hormone receptors

Miyashita

Matsushita

Nakamura

et al. 2011

Insect Biochemistry and Molecular Biology

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Recent advances in mass spectrometry (MS) technology have facilitated the detection and quantification of minor components in organisms and the environment. In this study, we successfully identified 20-hydroxyecdysone (20E) in first instar nymphs (7 days after hatching) of the scorpion Liocheles australasiae, using tandem mass spectrometry combined with high-performance liquid chromatography (LC/MS/MS). This substance was not found in adults after the fifth stage. Other possible molting hormone candidates such as makisterone A (MaA) and ponasterone A (PoA), both of which are reported to be the molting hormones of a few arthropod species, were not detected in this scorpion. The ligand-receptor binding of 20E and its analogs was quantitatively evaluated against the in vitro-translated molting hormone receptor, the heterodimer of ecdysone receptor (EcR) and the retinoid X receptor (RXR) of L. australasiae (LaEcR/LaRXR). The concentrations of ecdysone (E), MaA, 20E, and PoA that are required to inhibit 50% of [(3)H]PoA binding to the LaEcR/LaRXR complex were determined to be 1.9, 0.69, 0.05, and 0.017 μM, respectively. The activity profiles of these 4 ecdysteroids are consistent with those obtained for the molting hormone receptors of several insects. The binding of a non-steroidal E agonist, tebufenozide, to EcR was not observed even at high concentrations, indicating that the structure of the ligand-binding pocket of LaEcR is not favorable for interaction with tebufenozide.

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Section: Binding Affinity Of Ecdysteroids To Ecrsmentioning

confidence: 97%

LC/MS/MS identification of 20-hydroxyecdysone in a scorpion (Liocheles australasiae) and its binding affinity to in vitro-translated molting hormone receptors

Miyashita

Matsushita

Nakamura

et al. 2011

Insect Biochemistry and Molecular Biology

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“…Until the present study, full-length ecdysone receptor heterodimers had been cloned, and recombinant protein characterised, from only two members of the Hemiptera, namely B. tabaci and M. persicae (Carmichael et al, 2005;Graham et al, 2007a, b;Graham et al, 2009); both belong to the suborder Sternorrhyncha. The cloning, expression and characterisation of the N. viridula ecdysone receptor, reported in this account, provides the first access to a recombinant ecdysone receptor protein from the hemipteran suborder Heteroptera.…”

Section: Phylogeny Of the N Viridula Ecdysone Receptormentioning

confidence: 99%

“…A cDNA library (1 Â 10 6 pfu) produced from late fifth-instar N. viridula nymphs was amplified to 9.4 Â 10 8 pfu and screened, essentially as described by Graham et al (2009). Positive isolate pBK-CMV phagemids NvEcR10, NvEcR11 and NvUSP were selected based on size of insert and 5 0 and 3 0 sequence.…”

Section: Cdna Library Construction and Cloning Of Nvecrs And Nvuspmentioning

confidence: 99%

“…However, much of what is known of insect ecdysone receptors has been derived from studies on dipteran and lepidopteran species. It was not until relatively recently that the first ecdysone receptor-encoding sequences were identified/characterised in hemipteran species: Bemisia tabaci (Carmichael et al, 2005;Graham et al, 2007a, b;Graham et al, 2009), Myzus persicae (Graham et al, 2007a, b;Graham et al, 2009) and Acyrthosiphon pisum (Sabater-Munoz et al, 2006). All three of these species belong to the suborder Sternorrhyncha.…”

Section: Introductionmentioning

confidence: 96%

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An ecdysone receptor from the pentatomomorphan, Nezara viridula, shows similar affinities for moulting hormones makisterone A and 20-hydroxyecdysone

Tohidi-Esfahani

Graham

Hannan

et al. 2011

Insect Biochemistry and Molecular Biology

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“…Recent experiments (Carmichael et al ., 2005; Graham et al ., 2007) have revealed that the binding affinities of tebufenozide (a DBH compound) vary greatly across the taxonomic orders of arthropods and reflect lepidopteran‐selective toxicity. Additionally, in vitro assays of the binding affinities of tebufenozide have shown a strong negative correlation with the phylogenetic distances of the EcR‐LBDs from a lepidopteran reference sequence (Graham et al ., 2009). This has attracted attention because the moulting process of most insects is regulated by the steroidal hormone 20E.…”

Section: Introductionmentioning

confidence: 99%

Structural changes under low evolutionary constraint may decrease the affinity of dibenzoylhydrazine insecticides for the ecdysone receptor in non‐lepidopteran insects

Zotti

Christiaens

Rougé

et al. 2012

Insect Molecular Biology

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Understanding how variations in genetic sequences are conveyed into structural and biochemical properties is of increasing interest in the field of molecular evolution. In order to gain insight into this process, we studied the ecdysone receptor (EcR), a transcription factor that controls moulting and metamorphosis in arthropods. Using an in silico homology model, we identified a region in the lepidopteran EcR that has no direct interaction with the natural hormone but is under strong evolutionary constraint. This region causes a small indentation in the three-dimensional structure of the protein which facilitates the binding of tebufenozide. Non-Mecopterida are considered much older, evolutionarily, than Lepidoptera and they do not have this extended cavity. This location shows differences in evolutionary constraint between Lepidoptera and other insects, where a much lower constraint is observed compared with the Lepidoptera. It is possible that the higher flexibility seen in the EcR of Lepidoptera is an entirely new trait and the higher constraint could then be an indication that this region does have another important function. Finally, we suggest that Try123, which is evolutionarily constrained and is up to now exclusively present in Lepidoptera EcRs, could play a critical role in discriminating between steroidal and non-steroidal ligands.

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Ecdysone Receptors of Pest Insects–Molecular Cloning, Characterisation, and a Ligand Binding Domain-Based Fluorescence Polarization Screen

Cited by 6 publications

References 72 publications

LC/MS/MS identification of 20-hydroxyecdysone in a scorpion (Liocheles australasiae) and its binding affinity to in vitro-translated molting hormone receptors

LC/MS/MS identification of 20-hydroxyecdysone in a scorpion (Liocheles australasiae) and its binding affinity to in vitro-translated molting hormone receptors

An ecdysone receptor from the pentatomomorphan, Nezara viridula, shows similar affinities for moulting hormones makisterone A and 20-hydroxyecdysone

Structural changes under low evolutionary constraint may decrease the affinity of dibenzoylhydrazine insecticides for the ecdysone receptor in non‐lepidopteran insects

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