Sammelbandbeitrag

Erschienen in: HDAC/HAT Function Assessment and Inhibitor Development. Methods and Protocols

Herausgeberliste: Krämer, O.H.

Jahr der Veröffentlichung: 2023

Seiten: 361-376

ISBN: 978-1-0716-2787-7

eISBN: 978-1-0716-2788-4

DOI Link: https://doi.org/10.1007/978-1-0716-2788-4_24

Serientitel: Methods in Molecular Biology

Serienzählung: 2589

Abstract: 
Experiments determining the chromatin association of histone acetylases (HATs) and deacetylases (HDACs) at the genome-wide level provide precise maps of locus occupancy, but do not allow conclusions on the functional consequences of this locus-specific enrichment. Here we describe a protocol that allows tethering of HATs or HDACs to specific genomic loci upon fusion with a fluorescent protein and a DNA-binding protein such as the E. coli Lac repressor (LacI), which binds to genomically inserted lac operon sequences (lacO) via DNA/protein interactions. Integration of these lacO sequences into a genomic region of interest allows to monitor the functional consequences of HAT/HDAC targeting on chromatin (de)compaction, histone modification, and interaction with other proteins by quantitative light microscopy, as described here. As DNA-binding of LacI can be tightly controlled by the addition of galactose-derivatives, this method also allows to monitor the effects of locus-specific recruitment in a time-resolved manner.

Harvard-Zitierstil: Pfisterer, M. and Schmitz, M. (2023) Testing the Effect of Histone Acetyltransferases on Local Chromatin Compaction, in Krämer, O. (ed.) HDAC/HAT Function Assessment and Inhibitor Development. Methods and Protocols. New York, NY: Humana, pp. 361-376. https://doi.org/10.1007/978-1-0716-2788-4_24

APA-Zitierstil: Pfisterer, M., & Schmitz, M. (2023). Testing the Effect of Histone Acetyltransferases on Local Chromatin Compaction. In Krämer, O. (Ed.), HDAC/HAT Function Assessment and Inhibitor Development. Methods and Protocols (pp. 361-376). Humana. https://doi.org/10.1007/978-1-0716-2788-4_24