Abstract
[3H]Ponasterone A (PNA) of high specific activity has been used to identify and begin characterization of ecdysteroid (formerly called ecdysone) receptors in cytoplasmic and nuclear fractions of imaginal discs of Drosophila melanogaster. The equilibrium Kd of the observed macromolecular binding, 3--4 X 10(-9) M PNA, is in good agreement with the minimal concentration required for induction of complete morphogenesis in vitro, 4.2 X 10(-9) M PNA. Binding is analog specific and has kinetics consistent with a role in hormone response. On gentle homogenization, less than 5% of the binding capacity of the cell is released as soluble receptor; the other 95% remains with the nuclear fraction. This nuclear fraction specifically binds [3H]PNA in vitro. Greater than 95% of nuclear PNA receptors are released by extraction with 0.3 M KCl. The binding properties of the nuclear receptors are indistinguishable from those of the cytosol fraction or of the whole cell.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson L. E., McClure W. O. An improved scintillation cocktail of high-solubilizing power. Anal Biochem. 1973 Jan;51(1):173–179. doi: 10.1016/0003-2697(73)90465-x. [DOI] [PubMed] [Google Scholar]
- Beadle G. W., Ephrussi B. The Differentiation of Eye Pigments in Drosophila as Studied by Transplantation. Genetics. 1936 May;21(3):225–247. doi: 10.1093/genetics/21.3.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borst D. W., Bollenbacher W. E., O'Connor J. D., King D. S., Fristrom J. W. Ecdysone levels during metamorphosis of Drosophila melanogaster. Dev Biol. 1974 Aug;39(2):308–316. doi: 10.1016/0012-1606(74)90242-5. [DOI] [PubMed] [Google Scholar]
- Chang E. S., Sage B. A., O'Connor J. D. The qualitative and quantitative determinations of ecdysones in tissues of the crab, Pachygrapsus crassipes, following molt induction. Gen Comp Endocrinol. 1976 Sep;30(1):21–33. doi: 10.1016/0016-6480(76)90062-9. [DOI] [PubMed] [Google Scholar]
- Chihara C. J., Petri W. H., Fristrom J. W., King D. S. The assay of ecdysones and juvenile hormones on Drosophila imaginal disks in vitro. J Insect Physiol. 1972 Jun;18(6):1115–1123. doi: 10.1016/0022-1910(72)90146-1. [DOI] [PubMed] [Google Scholar]
- Fristrom J. W., Gregg T. L., Siegel J. The effect of beta-ecdysone on protein synthesis in imaginal discs of Drosophila melanogaster cultured in vitro. Dev Biol. 1974 Dec;41(2):301–313. doi: 10.1016/0012-1606(74)90308-x. [DOI] [PubMed] [Google Scholar]
- Fristrom J. W., Logan W. R., Murphy C. The synthetic and minimal culture requirements for evagination of imaginal discs of Drosophila melanogaster in vitro. Dev Biol. 1973 Aug;33(2):441–456. doi: 10.1016/0012-1606(73)90149-8. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Maroy P., Dennis R., Beckers C., Sage B. A., O'Connor J. D. Demonstration of an ecdysteroid receptor in a cultured cell line of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6035–6038. doi: 10.1073/pnas.75.12.6035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nishiura J. T., Fristrom J. W. Effect of insect hormones on RNA polymerases of mass-isolated imaginal discs of Drosophila melanogaster cultured in vitro. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2984–2988. doi: 10.1073/pnas.72.8.2984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petri W. H., Fristrom J. W., Stewart D. J., Hanly E. W. The in vitro synthesis and characteristics of ribosomal RNA in imaginal discs of Drosophila melanogaster. Mol Gen Genet. 1971;110(3):245–262. doi: 10.1007/BF00337837. [DOI] [PubMed] [Google Scholar]
- Raikow R., Fristrom J. W. Effects of -ecdysone on RNA metabolism of imaginal disks of Drosophila melanogaster. J Insect Physiol. 1971 Sep;17(9):1599–1614. doi: 10.1016/0022-1910(71)90056-4. [DOI] [PubMed] [Google Scholar]
- Robb J. A. Maintenance of imaginal discs of Drosophila melanogaster in chemically defined media. J Cell Biol. 1969 Jun;41(3):876–885. doi: 10.1083/jcb.41.3.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schooley D. A., Weiss G., Nakanishi K. A simple and general extraction procedure for phytoecdysones based on reversed-phase adsorption chromatography. Steroids. 1972 Mar;19(3):377–383. doi: 10.1016/0039-128x(72)90077-3. [DOI] [PubMed] [Google Scholar]
- Siegel J. G., Fristrom J. W. The effect of beta-ecdysone on protein synthesis in imaginal discs of Drosophila melanogaster cultured in vitro. Dev Biol. 1974 Dec;41(2):314–330. doi: 10.1016/0012-1606(74)90309-1. [DOI] [PubMed] [Google Scholar]
- Williams D., Gorski J. A new assessment of subcellular distribution of bound estrogen in the uterus. Biochem Biophys Res Commun. 1971 Oct 1;45(1):258–264. doi: 10.1016/0006-291x(71)90078-7. [DOI] [PubMed] [Google Scholar]
- Yund M. A., Fristrom J. W. Uptake and binding of beta-ecdysone in imaginal discs of Drosophila melanogaster. Dev Biol. 1975 Apr;43(2):287–298. doi: 10.1016/0012-1606(75)90028-7. [DOI] [PubMed] [Google Scholar]