MPP8 is essential for sustaining self-renewal of ground-state pluripotent stem cells

Description: Summary from the GEO: "Deciphering the mechanisms that control the pluripotent ground state is key for understanding embryonic development. Nonetheless, the epigenetic regulation of ground-state mouse embryonic stem cells (mESCs) is not fully understood. Here, we identify the epigentic protein MPP8 as being essential for ground-state pluripotency. Its depletion leads to cell cycle arrest and spontaneous differentiation. MPP8 has been suggested to repress LINE1 elements by recruiting the human silencing hub (HUSH) complex to H3K9me3-rich regions. Unexpectedly, we find that LINE1 elements are efficiently repressed by MPP8 lacking the chromodomain, while the unannotated C-terminus is essential for its function. Moreover, we show that SETDB1 recruits MPP8 to its genomic target loci, whereas transcriptional repression of LINE1 elements is maintained without retaining H3K9me3 levels. Taken together our findings demonstrate that MPP8 protects the DNA-hypomethylated pluripotent ground state through its association with the HUSH core complex, however, independently of detectable chromatin binding and maintenance of H3K9me3. "

Overall design from the GEO: "MPP8, FLAG and H3K9me3 chromatin immunoprecipitation profiles (ChIP-seq) analyzed in mouse embryonic stem cells which have been engineered to homozygously express MPP8 carrying a C-terminal miniAID (mAID) tag from the endogenous Mphosph8 locus as well as the E3 ligase OsTIR1 exogenously (Mpp8mAID; OsTIR1 cells). Cells were left untreated or treated with auxin (IAA) for indicated time points to remove endogenous MPP8. MPP8, FLAG and H3K9me3 chromatin immunoprecipitation profiles (ChIP-seq) analyzed in the Mpp8mAID; OsTIR1 background additionally expressing FLAG-tagged MPP8 fragments, Mpp8wt, Mpp8112-858 and MPP81-729, respectively (wt corresponds to the full length protein sequence, Npart corresponds to amino acid 1-729, Cpart correspond to amino acid 112-858). Cells were left untreated or treated with auxin (IAA) for indicated time points to remove endogenous MPP8. FLAG ChIPSeq analysis in cells engineered to homozygously express MPP8, TASOR and PPHLN1 carrying a C-terminal FLAG2 tag at the endogenous Mphosph8, Tasor and Pphln1 loci, respectively, or untagged parental control. RNA-seq analysis in Mpp8mAID; OsTIR1 cells, and cells additionally expressing FLAG-tagged MPP8 fragments, Mpp8wt, Mpp8112-858 and MPP81-729, respectively. Cells were left untreated or treated with auxin (IAA) for indicated time points to remove endogenous MPP8. "

Subject of Study

Mus musculus

Subject(s)

Gene Knock-In Techniques

Mouse Embryonic Stem Cells

Phosphoproteins

Pluripotent Stem Cells

Access via GEO
Plaintext, BED and BW of RNA and ChIP sequencing
Accession #: GSE150926

Access via SRA
RNA and ChIP sequencing of 110 samples
Accession #: SRP262463

Access via BioProject
Additional information about overall initiative.
Accession #: PRJNA634099
Access Restrictions: Free to All
Access Instructions: The NCBI Gene Expression Omnibus, SRA, and BioProject databases provide open access to these files.
Associated Publications: Müller I, Moroni AS, Shlyueva D, et al. MPP8 is essential for sustaining self-renewal of ground-state pluripotent stem cells. Nat Commun. 2021;12(1):3034. Published 2021 May 24.
Data Type: Genomic
Equipment Used: Illumina NextSeq 500
Software Used: SRA Toolkit
The NCBI SRA Toolkit enables reading ("dumping") of sequencing files from the SRA database and writing ("loading") files into the .sra format.
Dataset Format(s): Plain Text, BED, BIGWIG, gzip
Data Tool(s): RNA Seq

ChIP Seq
Dataset Size: 1.4 KB (BED), 9.0 GB (TAR of BW), 15 MB (TXT)

Data Catalog Record Updated: 2023-12-07