Cloning of an ecdysone receptor homolog from Manduca sexta and the developmental profile of its mRNA in Wings

Fujiwara, Haruhiko; Jindra, Marek; Newitt, Richard; Palli, Subba Reddy; Hiruma, Kiyoshi; Riddiford, Lynn M.

doi:10.1016/0965-1748(95)00023-o

Cited by 175 publications

(97 citation statements)

References 43 publications

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“…The molecular basis of 20E action has been elucidated in great detail (10,24,26,46,51,56,64,65). The two nuclear hormone receptors that play a central role in the initiation of the 20E-induced gene regulatory hierarchy are the ecdysteroid receptor (EcR) and the retinoid X receptor (RXR) homologue Ultraspiracle (USP) (12,18,25,27,34,43,57,60). It has been shown that USP is an obligatory heterodimeric partner of EcR, required for both ligand and DNA binding (63,74,75).…”

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confidence: 99%

Molecular Determinants of Differential Ligand Sensitivities of Insect Ecdysteroid Receptors

Wang

Ayer

Segraves

et al. 2000

Molecular and Cellular Biology

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The functional receptor for insect ecdysteroid hormones is a heterodimer consisting of two nuclear hormone receptors, ecdysteroid receptor (EcR) and the retinoid X receptor homologue Ultraspiracle (USP). Although ecdysone is commonly thought to be a hormone precursor and 20-hydroxyecdysone (20E), the physiologically active steroid, little is known about the relative activity of ecdysteroids in various arthropods. As a step toward characterization of potential differential ligand recognition, we have analyzed the activities of various ecdysteroids using gel mobility shift assays and transfection assays in Schneider-2 (S2) cells. Ecdysone showed little activation of the Drosophila melanogaster receptor complex (DmEcR-USP). In contrast, this steroid functioned as a potent ligand for the mosquito Aedes aegypti receptor complex (AaEcR-USP), significantly enhancing DNA binding and transactivating a reporter gene in S2 cells. The mosquito receptor also displayed higher hormoneindependent DNA binding activity than the Drosophila receptor. Subunit-swapping experiments indicated that the EcR protein, not the USP protein, was responsible for ligand specificity. Using domain-swapping techniques, we made a series of Aedes and Drosophila EcR chimeric constructs. Differential ligand responsiveness was mapped near the C terminus of the ligand binding domain, within the identity box previously implicated in the dimerization specificity of nuclear receptors. This region includes helices 9 and 10, as determined by comparison with available crystal structures obtained from other nuclear receptors. Site-directed mutagenesis revealed that Phe529 in Aedes EcR, corresponding to Tyr611 in Drosophila EcR, was most critical for ligand specificity and hormone-independent DNA binding activity. These results demonstrated that ecdysone could function as a bona fide ligand in a species-specific manner.

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confidence: 99%

Molecular Determinants of Differential Ligand Sensitivities of Insect Ecdysteroid Receptors

Wang

Ayer

Segraves

et al. 2000

Molecular and Cellular Biology

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“…The difference in EcR or USP isoforms is generally created by the link of different exons in the A/B regions. In the case of H. axyridis and E. vigintioctopunctata, the A/B regions of EcRB1s are longer than those of EcR-As, as reported in Bombyx mori, 35,36) Manduca sexta, 37,38) and Drosophila melanogaster. 39,40) A/B regions are homologous between B1 isoforms (89% identity for HaEcR-B1/EvEcR-B1) and between A isoforms (80% identity for HaEcR-A/EvEcR-A) irrespective of the Coccinellidae species.…”

Section: Primary Sequences Of Lady Beetle Ecrsmentioning

confidence: 80%

cDNA cloning of ecdysone receptor (EcR) and ultraspiracle (USP) from Harmonia axyridis and Epilachna vigintioctopunctata and the evaluation of the binding affinity of ecdysone agonists to the in vitro translated EcR/USP heterodimers

et al. 2014

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“…Differential sensitivity due to differential receptor expression, such as occurs in the tobacco hornworm (Manduca sexta) where ecdysterone sensitivity is regulated via variable receptor expression (Fujiwara et al, 1995), cannot apply to gregarization in the locust since solitarious eggs are capable of responding to gregarizing cues (e.g. Figs·1, 2).…”

Section: Discussionmentioning

confidence: 99%

Swarm formation in the desert locust Schistocerca gregaria:isolation and NMR analysis of the primary maternal gregarizing agent

Miller

Islam

Claridge

et al. 2008

Journal of Experimental Biology

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SUMMARYDepending on their rearing density, female desert locusts Schistocerca gregaria epigenetically endow their offspring with differing phenotypes. To identify the chemical basis for such maternal transmission, we compared solitarious and gregarious locust egg pod foam using high performance liquid chromatography (HPLC). We revealed causal relationships between foam chemistry and hatchling phenotype (phase state) by iteratively applying HPLC fractions from gregarious locust egg foam extracts to solitarious eggs and assessing resulting hatchlings with a behavioural bioassay involving logistic regression. Selection and application of increasingly specific HPLC fractions allowed us to isolate compounds with gregarizing properties. Hatchling gregarization was triggered only by certain fractions and was dose dependent. In a final series of experiments, we characterized the most specific gregarizing fraction by nuclear magnetic resonance (NMR) spectroscopy. Here we present tentative structural features of the primary locust maternal gregarizing agent, which appears to be an alkylated L-dopa analogue. In addition, we propose a mechanism for phase-dependent regulation of this compoundʼs activity.

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Cloning of an ecdysone receptor homolog from Manduca sexta and the developmental profile of its mRNA in Wings

Cited by 175 publications

References 43 publications

Molecular Determinants of Differential Ligand Sensitivities of Insect Ecdysteroid Receptors

Molecular Determinants of Differential Ligand Sensitivities of Insect Ecdysteroid Receptors

cDNA cloning of <i>ecdysone receptor</i> (<i>EcR</i>) and <i>ultraspiracle</i> (<i>USP</i>) from <i>Harmonia axyridis</i> and <i>Epilachna vigintioctopunctata</i> and the evaluation of the binding affinity of ecdysone agonists to the <i>in vitro</i> translated EcR/USP heterodimers

Swarm formation in the desert locust Schistocerca gregaria:isolation and NMR analysis of the primary maternal gregarizing agent

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