Glucocorticoid Hormone-Induced Receptor Localization to the Chromatin Fibers Formed on Injected DNA in Xenopus Oocytes
| Autor: | Örjan Wrange, Birgitta Gelius |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Tissue Fixation
Microinjections viruses Immunoblotting Active Transport Cell Nucleus Xenopus RNA polymerase II Biology Chromatin remodeling Mice Xenopus laevis chemistry.chemical_compound Receptors Glucocorticoid Glucocorticoid receptor Transcription (biology) Animals RNA Messenger Promoter Regions Genetic Glucocorticoids Cell Nucleus DNA Helicases Nuclear Proteins Cell Biology biology.organism_classification Molecular biology Chromatin Recombinant Proteins SWI/SNF DNA-Binding Proteins NFI Transcription Factors Mammary Tumor Virus Mouse Microscopy Fluorescence chemistry CCAAT-Enhancer-Binding Proteins Oocytes biology.protein RNA Polymerase II Y-Box-Binding Protein 1 DNA Signal Transduction Transcription Factors |
| Zdroj: | Experimental Cell Research. 265:319-328 |
| ISSN: | 0014-4827 |
| DOI: | 10.1006/excr.2001.5184 |
| Popis: | Oocytes from Xenopus laevis have provided a model system for studying the dynamic changes that occur in chromatin during gene activation. We have reconstituted glucocorticoid receptor (GR) induced transcription from the mouse mammary tumor virus (MMTV) promoter by intranuclear injection of an MMTV-driven reporter and cytoplasmic injection of synthetic mRNA GR into Xenopus oocytes. Here we investigate the intranuclear distribution of injected DNA, which is assembled into chromatin. We show that this chromatin is organized as an intranuclear fibrous network. Unliganded GR is located in the cytosol and hormone triggers its nuclear translocation and association with the chromatin fibers. Furthermore, we analyze the intranuclear distribution of other factors involved in transcription from the MMTV promoter. Indirect immunofluorescence microscopy on cryostat-sectioned oocytes revealed that BRG1, which is a subunit of the SWI/SNF chromatin remodeling complex, as well as RNA polymerase II and recombinantly expressed Xenopus nuclear factor 1-B, are all associated with the endogenous chromosomes and the chromatin fibers formed on injected DNA. This association does not depend on specific DNA binding sites and appears to be nonspecific. |
| Databáze: | OpenAIRE |
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