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Mechanisms of stimulation of SAGA-mediated nucleosome acetylation by a transcriptional activator

ABSTRACT: Eukaryotic gene expression requires the coordination of multiple factors to overcome the repressive nature of chromatin. However, the mechanistic details of this coordination are not well understood. The SAGA family of transcriptional coactivators interacts with DNA-binding activators to establish regions of hyperacetylation. We have previously shown that, contrary to the prevailing model in which activator protein increases SAGA affinity for nucleosome substrate, the Gal4-VP16 activator model system augments the rate of acetylation turnover for the SAGA complex from budding yeast. To better understand how this stimulation occurs, we have identified necessary components using both kinetics assays and binding interactions studies. We find that Gal4-VP16-mediated stimulation requires activator binding to DNA flanking the nucleosome, as it cannot be reproduced in trans by activator protein alone or by exogenous DNA containing the activator binding site in combination with the activator protein. Further, activator-mediated stimulation requires subunits outside of the histone acetylation (HAT) module, with the Tra1 subunit being responsible for the majority of the stimulation. Interestingly, for the HAT module alone, nucleosome acetylation is inhibited by activator proteins due to non-specific binding of the activator to the nucleosomes. This inhibition is not observed for the yeast ADA complex, a small complex comprised mostly of the HAT module, suggesting that subunits outside of the HAT module in both it and SAGA can overcome non-specific activator binding to nucleosomes. However, this activity appears distinct from activator-mediated stimulation, as ADA complex acetylation is not stimulated by Gal4-VP16. Graphical abstract Image 1 Highlights • Stimulation of nucleosome acetylation by SAGA requires activator in cis• Tra1 mediates the majority of activator stimulation• The core HAT complex of SAGA is inhibited by activator due to non-specific binding• The related ADA complex is neither stimulated nor inhibited by activator

SUBMITTER: Culbertson S

PROVIDER: S-EPMC7786029 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Mechanisms of stimulation of SAGA-mediated nucleosome acetylation by a transcriptional activator

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