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Context-dependent effects of Acf1 assessed in high throughput

ABSTRACT: Context-dependent effects of Acf1 assessed in high throughput

PROVIDER: PRJNA417865 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6279390.fastq.gz	Fastqsanger.gz
	SRR6279400.fastq.gz	Fastqsanger.gz
	SRR6279391.fastq.gz	Fastqsanger.gz
	SRR6279401.fastq.gz	Fastqsanger.gz
	SRR6279392.fastq.gz	Fastqsanger.gz

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Context-dependent effects of Acf1 assessed in high throughput

Project description:We applied a multiplexed genomic tethering assay (Brueckner, Epigenetics Chromatin, 2016; PMID:27777628) to elucidate the function of ACF1, the large subunit of the ACF nucleosome sliding complex in drosophila Kc167 cells. Targeting a Gal4DBD-ACF1 fusion to several hundred integrated reporters revealed a context-dependent repressive effect which is modulated by gene activity and chromatin state.

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Context-dependent effects of HP1 assessed in high throughput

Project description:Chromatin proteins control gene activity in a concerted manner. We developed a high-throughput assay to study the effects of the local chromatin environment on the regulatory activity of a protein of interest. The assay combines a previously reported multiplexing strategy based on barcoded randomly integrated reporter genes (Akhtar, Cell, 2013; PMID: 23953119) with Gal4-mediated tethering of the protein of interest. We applied the assay to Drosophila Heterochromatin protein 1a (HP1a), which is mostly known as a repressive protein working by a chromatin compaction mechanism but has also been linked to transcriptional activation. Probing the effects of HP1a in more than one thousand genomic locations revealed that HP1a is a potent repressor able to silence even highly expressing loci. However, the local chromatin context can modulate HP1a function. In pericentromeric regions, HP1a-induced repression was enhanced by two-fold. In regions marked by a H3K36me3-rich chromatin signature linked to transcriptional elongation, HP1a-dependent silencing was significantly decreased. We found no evidence for an activating function of HP1a in our experimental system. Furthermore, we did not observe stable transmission of repression over mitotic divisions after loss of targeted HP1a recruitment in any of the loci examined. The multiplexed tethered reporter assay will be a useful tool to dissect combinatorial regulatory interactions in chromatin.

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