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Updated: May 12, 2025

Immunofluorescent Staining for Visualization of Heterochromatin Associated Proteins in Drosophila Salivary Glands
Published on: August 21, 2021
Transcriptional induction by ecdysone in Drosophila salivary glands involves an increase in chromatin accessibility
Aleksandra A Evdokimova1, Tatyana D Kolesnikova2, Marina Yu Mazina1
1Institute of Gene Biology, Russian Academy of Sciences, 119334, Moscow, Russia.
Abstract:
Transcriptional activation by 20-hydroxyecdysone (20E) in Drosophila provides an excellent model for studying tissue-specific responses to steroids. An increase in the 20E concentration regulates the degradation of larval and the proliferation of adult tissues during metamorphosis. To study 20E-dependent transcription, we used the natural system for controlling the 20E concentration-the E23 membrane transporter-which exports 20E from the cell. We artificially expressed E23 in tissues to suppress the first wave of 20E-inducible transcription at metamorphosis. E23 expression revealed a plethora of 20E-dependent genes in salivary glands, while mildly affecting transcription in brain. We described the mechanisms controlling transcriptional activation by 20E in salivary glands. 20E depletion decreased the binding of Pol II and the TFIID subunit, TBP, to the promoters of primary targets, demonstrating the role of 20E in transcription initiation. At target loci, 20E depletion resulted in the malfunctioning of sites co-bound with EcR and CBP/Nejire and enriched for the H3K27Ac mark inherent to active enhancers. At these sites, the 20E concentration was found to control chromatin accessibility and acetylation. We suggest that the activity of these 'active' ecdysone-sensitive elements was responsible for the active status of 20E targets in the salivary glands of wandering larvae.
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