Creation of ecdysone receptor chimeras in plants for controlled regulation of gene expression

Martinez, Alberto; Sparks, Caroline A.; Drayton, Paul; Thompson, John A.; Greenland, Andrew J.; Jepson, Ian

doi:10.1007/s004380050999

Cited by 45 publications

(32 citation statements)

References 43 publications

(68 reference statements)

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“…2). Similar results have been reported in relation to recombinant transcription factors consisting of the ecdysone receptor-ligand binding domain, the GR TAD and its DNA-binding domain (Martinez et al 1999a). The VP16 TAD can directly interact with general transcription factors evolutionarily conserved among eukaryotes (Lin et al 1991;Shen et al 1996) and was also potent in plant cells (Schwechheimer et al 1998).…”

Section: Discussionsupporting

confidence: 79%

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants

Kodama

Okada

Inui

et al. 2007

Planta

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In mammals, the aryl hydrocarbon receptor (AhR) mediates expression of certain genes, including CYP1A1, in response to exposure to dioxins and related compounds. We have constructed a mouse AhR-mediated gene expression systems for a beta-glucuronidase (GUS) reporter gene consisting of an AhR, an AhR nuclear translocator (Arnt), and a xenobiotic response element (XRE)-driven promoter in transgenic tobacco plants. On treatment with the AhR ligands 3-methylcholanthrene (MC), beta-naphthoflavone (betaNF), and indigo, the transgenic tobacco plants exhibited enhanced GUS activity, presumably by inducible expression of the reporter gene. The recombinant AhR (AhRV), with the activation domain replaced by that of the Herpes simplex virus protein VP16, induced GUS activity much more than the wild-type AhR in the transgenic tobacco plants. Plants carrying AhRV expressed the GUS reporter gene in a dose- and time-dependent manner when treated with MC; GUS activity was detected at 5 nM MC on solid medium and at 12 h after soaking in 25 microM MC. Histochemical GUS staining showed that this system was active mainly in leaf and stem. These results suggest that the AhR-mediated reporter gene expression system has potential for the bioassay of dioxins in the environment and as a novel gene expression system in plants.

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Section: Discussionsupporting

confidence: 79%

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants

Kodama

Okada

Inui

et al. 2007

Planta

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“…A. Hansen, and S. Meyer, submission to GenBank, accession no. AF325360), lepidopteran (55)(56)(57)(58)(59), and coleopteran (60) EcRs.…”

Section: Intrinsic Activation Functions Of the Isoformspecific Amino mentioning

confidence: 99%

Transcription Activation by the Ecdysone Receptor (EcR/USP): Identification of Activation Functions

Cherbas

2003

Molecular Endocrinology

172

132

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The ecdysone receptor is a heterodimer of the two nuclear receptors EcR and ultraspiracle (USP). We have identified the regions of Drosophila EcR and USP responsible for transcriptional activation of a semisynthetic Eip71CD promoter in Kc cells. The isoform-specific A/B domains of EcR-B1 and B2, but not those of EcR-A or USP, exhibit strong activation activity [activation function 1 (AF1)], both in isolation and in the context of the intact receptor. AF1 activity in isoform B1 derives from dispersed elements; the B2-specific AF1 consists of a 17-residue amphipathic helix. AF2 function was studied using a two-hybrid assay in Kc cells, based on the observation that potent hormone-dependent activation by the EcR/USP ligand-binding domain heterodimer requires the participation of both partners. Mutagenesis reveals that AF2 function depends on EcR helix 12, but not on the cognate USP region. EcR helix 12 mutants (F645A and W650A) exhibit a dominant negative phenotype. Thus, in the setting tested, the ecdysone receptor can activate transcription using the AF1 regions of EcR-B1 or -B2 and the AF2 region of EcR. USP acts as an allosteric effector for EcR, but does not contribute any intrinsic function.

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“…Insecticide-induced gene expression may be a preferred scenario in plants since this technology would enable combination of pesticide treatment with expression of transgenic determinants. Recently, the Heliothis virescens ecdysone receptor (HVE) has been engineered for regulated gene expression in plants (Martinez et al, 1999a;Martinez et al, 1999b). Similar to mammalian configurations, HVE was fused to the VP16 transactivation domain and the DNA-binding domain of the mammalian glucocorticoid receptor.…”

Section: Ecdysone-responsive Expression Technologymentioning

confidence: 99%

The Impact of Mammalian Gene Regulation Concepts on Functional Genomic Research, Metabolic Engineering, and Advanced Gene Therapies

2001

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Regulation of heterologous gene expression is of prime importance for a wide variety of basic and applied biological research areas including functional genomics, tissue engineering, gene therapy, and biopharmaceutical manufacturing. Initial gene regulation strategies employed endogenous responsive elements, which resulted in pleiotropic interference of transgene expression with host regulatory networks. Current regulation systems are binary and consist of chimeric transactivators and responsive target promoters of heterologous bacterial or insect origin, or they contain artificially designed components. Regulation of generic systems is based on binding of a transactivator to its cognate promoter, which is modulated by specific molecules such as antibiotics or hormones and brings the transactivation domain into contact with a minimal promoter, thereby inducing target gene expression. Binary gene regulation concepts have been significantly refined in recent years with a focus to improve their regulation performance and their compatibility with human-therapeutic use. In this review we present a detailed analysis of currently available mammalian gene regulation systems and document progress that has pioneered the use of such systems in various aspects of human therapy.

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Creation of ecdysone receptor chimeras in plants for controlled regulation of gene expression

Cited by 45 publications

References 43 publications

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants

Aryl hydrocarbon receptor (AhR)-mediated reporter gene expression systems in transgenic tobacco plants

Transcription Activation by the Ecdysone Receptor (EcR/USP): Identification of Activation Functions

The Impact of Mammalian Gene Regulation Concepts on Functional Genomic Research, Metabolic Engineering, and Advanced Gene Therapies

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