Journal
EMBO JOURNAL
Volume 36, Issue 18, Pages 2710-2725Publisher
WILEY
DOI: 10.15252/embj.201696035
Keywords
diffusion constant; FCS; FLIP; FRAP; tudor domain
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Funding
- IGBMC PhD fellowship
- Fondation pour la Recherche Medicale (FRM) fellowship
- CNRS
- INSERM
- Strasbourg University
- EC Marie Curie-ITN (NR-NET)
- Agence Nationale de la Recherche [ANR-11-BSV5-010-02 Chromact, ANR-13-BSV6-0001-02 COREAC, ANR-13-BSV8-0021-03 DiscoverIID]
- European Research Council (ERC) Advanced grant (Birtoaction) [ERC-2013-340551]
- French State fund by Agence Nationale de la Recherche under the frame program Investissements d'Avenir [ANR-10-LABX-0030-INRT, ANR-10-IDEX-0002-02]
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SAGA and ATAC are two distinct chromatin modifying co-activator complexes with distinct enzymatic activities involved in RNA polymerase II (Pol II) transcription regulation. To investigate the mobility of co-activator complexes and general transcription factors in live-cell nuclei, we performed imaging experiments based on photobleaching. SAGA and ATAC, but also two general transcription factors (TFIID and TFIIB), were highly dynamic, exhibiting mainly transient associations with chromatin, contrary to Pol II, which formed more stable chromatin interactions. Fluorescence correlation spectroscopy analyses revealed that the mobile pool of the two co-activators, as well as that of TFIID and TFIIB, can be subdivided into fast (free) and slow (chromatin-interacting) populations. Inhibiting transcription elongation decreased H3K4 trimethylation and reduced the slow population of SAGA, ATAC, TFIIB and TFIID. In addition, inhibiting histone H3K4 trimethylation also reduced the slow populations of SAGA and ATAC. Thus, our results demonstrate that in the nuclei of live cells the equilibrium between fast and slow population of SAGA or ATAC complexes is regulated by active transcription via changes in the abundance of H3K4me3 on chromatin.
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