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Series GSE83714 Query DataSets for GSE83714
Status Public on Sep 20, 2016
Title Context-dependent effects of HP1 assessed in high throughput
Organism Drosophila melanogaster
Experiment type Expression profiling by high throughput sequencing
Other
Summary 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.
 
Overall design TRIP assay in drosophila Kc167 cells combined with targeted recruitment of Gal4-HP1 to integrated 5xGa4UAS-pMT-GFP reporters. Two technical replicate experiments of TRIP pool transfected with Gal4-HP1, Gal4 or HP1 performed by day 2 and day 16 after transient transfection.
 
Contributor(s) Brueckner L, van Arensbergen J, Akhtar W, Pagie L, van Steensel B
Citation(s) 27777628
Submission date Jun 24, 2016
Last update date May 15, 2019
Contact name Bas van Steensel
E-mail(s) b.v.steensel@nki.nl
Phone + 31 20 512 2040
Fax +31 20 669 1383
URL http://www.nki.nl/nkidep/vansteensel
Organization name Netherlands Cancer Institute
Department division of Molecular Biology
Lab van Steensel group
Street address Plesmanlaan 121
City Amsterdam
ZIP/Postal code 1066 CX
Country Netherlands
 
Platforms (3)
GPL13304 Illumina HiSeq 2000 (Drosophila melanogaster)
GPL16479 Illumina MiSeq (Drosophila melanogaster)
GPL17275 Illumina HiSeq 2500 (Drosophila melanogaster)
Samples (26)
GSM2213948 D2, cDNA reads, Gal4-HP1, repl 1
GSM2213949 D2, cDNA reads, Gal4-HP1, repl 2
GSM2213950 D2, cDNA reads, Gal4, repl 1
This SubSeries is part of SuperSeries:
GSE83715 HP1
Relations
BioProject PRJNA326782
SRA SRP077070

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE83714_TRIP_HP1_D16.csv.gz 69.5 Kb (ftp)(http) CSV
GSE83714_TRIP_HP1_D2.csv.gz 96.5 Kb (ftp)(http) CSV
GSE83714_spikein_BCDs.txt.gz 2.3 Kb (ftp)(http) TXT
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Raw data are available in SRA
Processed data are available on Series record

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