High level transactivation by a modified Bombyx ecdysone receptor in mammalian cells without exogenous retinoid X receptor

Our studies of the Bombyx mori ecdysone receptor (BE) revealed that, unlike the Drosophila melanogaster ecdysone receptor (DE), treatment of BE with the ecdysone agonist tebufenozide stimulated high level transactivation in mammalian cells without adding an exogenous heterodimer partner. Gel mobilit...

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Bibliographic Details
Published in:	Proceedings of the National Academy of Sciences - PNAS Vol. 95; no. 14; pp. 7999 - 8004
Main Authors:	Suhr, S.T. (The Salk Institute for Biological Studies, La Jolla, CA.), Gil, E.B, Senut, M.C, Gage, F.H
Format:	Journal Article
Language:	English
Published:	United States National Academy of Sciences of the United States of America 07-07-1998 National Acad Sciences National Academy of Sciences The National Academy of Sciences
Subjects:	ADN ADN RECOMBINADO ADN RECOMBINE Amino acids ANIMAL TRANSGENIQUE ANIMALES TRANSGENICOS Animals BINDING BINDING PROTEINS BINDING SITES Biochemistry BIOLOGICAL DIFFERENCES Biological Sciences Bombyx BOMBYX MORI CELL CULTURE Cell Line CELL LINES Cells Cellular biology Chimeras CULTIVO DE CELULAS CULTURE DE CELLULE DIFERENCIAS BIOLOGICAS DIFFERENCE BIOLOGIQUE Dimerization Dimers DNA DROSOPHILA MELANOGASTER Ecdysone - metabolism EXPRESION GENICA EXPRESSION DES GENES FACTEUR DE TRANSCRIPTION FACTORES DE TRANSCRIPCION Gels GENE EXPRESSION Genes HORMONAS DE LA MUDA HORMONE DE MUE HORMONE RECEPTORS Insects Ligands MAMIFEROS MAMMALS MAMMIFERE MOULTING HORMONES PROTEINAS AGLUTINANTES PROTEINE DE LIAISON Proteins RECEPTEUR D'HORMONE RECEPTORES DE HORMONAS Receptors Receptors, Retinoic Acid - genetics Receptors, Retinoic Acid - metabolism Receptors, Steroid - genetics Receptors, Steroid - metabolism RECOMBINANT DNA Retinoid X Receptors SPECIES DIFFERENCES TEBUFENOZIDE Transactivation TRANSCRIPTION FACTORS Transcription Factors - genetics Transcription Factors - metabolism Transcriptional Activation Transfection TRANSGENIC ANIMALS
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Summary:	Our studies of the Bombyx mori ecdysone receptor (BE) revealed that, unlike the Drosophila melanogaster ecdysone receptor (DE), treatment of BE with the ecdysone agonist tebufenozide stimulated high level transactivation in mammalian cells without adding an exogenous heterodimer partner. Gel mobility shift and transfection assays with both the ultraspiracle gene product (Usp) and retinoid X receptor heterodimer partners indicated that this property of BE stems from significantly augmented heterodimer complex formation and concomitant DNA binding. We have mapped this "gain of function" to determinants within the D and E domains of BE and demonstrated that, although the D domain determinant is sufficient for high affinity heterodimerization with Usp, both determinants are necessary for high affinity interaction with retinoid X receptor. Modified BE receptors alone used as replication-defective retroviruses potently stimulated separate "reporter" viruses in all cell types examined, suggesting that BE has potentially broad utility in the modulation of transgene expression in mammalian cells
Bibliography:	1999002857 L10 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 To whom reprint requests should be addressed. e-mail: Fgage@salk.edu. Communicated by Michael G. Rosenfeld, University of California at San Diego, La Jolla, CA
ISSN:	0027-8424 1091-6490
DOI:	10.1073/pnas.95.14.7999