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| Status |
Public on May 11, 2021 |
| Title |
ChIP-seq analysis of wild-type and K885A SETDB1 in 3T3-L1 preadipocytes |
| Organism |
Mus musculus |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
The lysine methyltransferase SETDB1, an enzyme responsible for methylation of histone H3 at lysine 9, plays a key role in H3K9 tri-methylation dependent silencing of endogenous retroviruses and developmental genes. Recent studies have shown that ubiquitination of human SETDB1 complements its catalytic activity and the silencing of endogenous retroviruses in human embryonic stem cells. However, it is not known whether SETDB1 ubiquitination is essential for its other major role in epigenetic silencing of developmental gene programs. We previously showed that SETDB1 contributes to the formation of H3K4/H3K9me3 bivalent chromatin domains that keep adipogenic Cebpa and Pparg genes in a poised state for activation and restricts the differentiation potential of pre-adipocytes. Here, we show that ubiquitin resistant K885A mutant of SETDB1 represses adipogenic genes and inhibits preadipocyte differentiation similar to wild-type SETDB1. We show this was due to a compensation mechanism for H3K9me3 chromatin modifications on the Cebpa locus by other H3K9 methyltransferases Suv39H1 and Suv39H2. In contrast, the K885A mutant did not repress other SETDB1 target genes such as Tril and Gas6 suggesting SETDB1 represses its target genes by two mechanisms; one that requires its ubiquitination and another that still requires SETDB1 but not its enzyme activity.
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| Overall design |
Retroviral expression of FLAG-tagged wild-type and K885A mutant mSETDB1 in 3T3-L1 preadipocytes. ChIP using anti-FLAG antibody followed by next-generation sequencing.
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| Contributor(s) |
Matsumura Y |
| Citation(s) |
33971063 |
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| Submission date |
Apr 26, 2021 |
| Last update date |
May 11, 2021 |
| Contact name |
Yoshihiro Matsumura |
| E-mail(s) |
ymatsumura@med.akita-u.ac.jp
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| Organization name |
Akita University Graduate School of Medicine
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| Department |
Biochemistry and Metabolic Science
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| Street address |
Hondo 1-1-1
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| City |
Akita |
| State/province |
Akita |
| ZIP/Postal code |
010-8543 |
| Country |
Japan |
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| Platforms (1) |
| GPL11002 |
Illumina Genome Analyzer IIx (Mus musculus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA725182 |
| SRA |
SRP316370 |
