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Status |
Public on Sep 07, 2021 |
Title |
Morc3 silences endogenous retroviruses by enabling Daxx-mediated H3.3 incorporation [ChIP-seq] |
Organism |
Mus musculus |
Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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Summary |
Endogenous retroviruses (ERVs) comprise a significant portion of mammalian genomes. Although, specific ERV loci feature regulatory roles for host gene expression, most ERV integrations are transcriptionally repressed by Setdb1 mediated H3K9me3 and DNA methylation. However, the protein network which regulates deposition of these chromatin modifications is still incompletely understood. Here, we performed a genome-wide sgRNA screen for genes involved in ERV silencing and identified the GHKL ATPase protein Morc3 as top scoring hit. Morc3 knock-out cells display de-repression, reduced H3K9me3 and increased chromatin accessibility of distinct ERV classes. We found that the GHKL ATPase domain of Morc3 is critical for ERV silencing, since mutants which cannot bind ATP, or which are defective in ATP hydrolysis cannot rescue the Morc3 ko phenotype. Proteomic analysis revealed that Morc3 mutant protein which cannot bind ATP fails to interact with the H3.3 chaperone Daxx. This interaction depends on Morc3 sumoylation, as Daxx lacking the SUMO interaction domain shows reduced association with Morc3. Notably, in Morc3 ko cells, we observed strongly reduced H3.3 on Morc3 binding sites. Thus, our data demonstrate Morc3 as critical regulator of Daxx-mediated H3.3 incorporation into ERV regions.
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Overall design |
ChIP analysis of the H3K4me3, H3K9me3 and H3K27ac histone modifications, H3.3 histone variant, Setdb1 and wild type and mutant Morc3 in different mouse embryonic stem cell lines
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Contributor(s) |
Groh S, de Almeida GP, Schotta G |
Citation(s) |
34650047 |
Submission date |
Oct 23, 2020 |
Last update date |
Nov 03, 2021 |
Contact name |
Gunnar Schotta |
E-mail(s) |
gunnar.schotta@bmc.med.lmu.de
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Phone |
089218075422
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Organization name |
LMU Munich
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Department |
Biomedical Center
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Street address |
Grosshaderner Strasse 9
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City |
Planegg-Martinsried |
ZIP/Postal code |
82152 |
Country |
Germany |
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Platforms (1) |
GPL18480 |
Illumina HiSeq 1500 (Mus musculus) |
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Samples (47)
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This SubSeries is part of SuperSeries: |
GSE159936 |
Morc3 silences endogenous retroviruses by enabling Daxx-mediated H3.3 incorporation |
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Relations |
BioProject |
PRJNA670920 |
SRA |
SRP288314 |