Eset partners with Oct4 to restrict extraembryonic trophoblast lineage potential in embryonic stem cells.
The histone H3 Lys 9 (H3K9) methyltransferase Eset is an epigenetic regulator critical for the development of the inner cell mass (ICM). Although ICM-derived embryonic stem (ES) cells are normally unable to contribute to the trophectoderm (TE) in blastocysts, we find that depletion of Eset by shRNAs leads to differentiation with the formation of trophoblast-like cells and induction of trophoblast-associated gene expression. Using chromatin immmunoprecipitation (ChIP) and sequencing (ChIP-seq) analyses, we identified Eset target genes with Eset-dependent H3K9 trimethylation. We confirmed that genes that are preferentially expressed in the TE (Tcfap2a and Cdx2) are bound and repressed by Eset. Single-cell PCR analysis shows that the expression of Cdx2 and Tcfap2a is also induced in Eset-depleted morula cells. Importantly, Eset-depleted cells can incorporate into the TE of a blastocyst and, subsequently, placental tissues. Coimmunoprecipitation and ChIP assays further demonstrate that Eset interacts with Oct4, which in turn recruits Eset to silence these trophoblast-associated genes. Our results suggest that Eset restricts the extraembryonic trophoblast lineage potential of pluripotent cells and links an epigenetic regulator to key cell fate decision through a pluripotency factor.
Yuan
Ping
P
Gene Regulation Laboratory, Genome Institute of Singapore, Singapore 138672.
Han
Jianyong
J
Guo
Guoji
G
Orlov
Yuriy L
YL
Huss
Mikael
M
Loh
Yuin-Han
YH
Yaw
Lai-Ping
LP
Robson
Paul
P
Lim
Bing
B
Ng
Huck-Hui
HH
D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D047369:Chromatin Immunoprecipitation; D053595:Embryonic Stem Cells / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D016678:Genome / Q000502:physiology; D011495:Histone-Lysine N-Methyltransferase; D018398:Homeodomain Proteins / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism*; D051379:Mice; D009028:Morula / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D011496:Protein Methyltransferases / Q000378:metabolism*; D050656:Transcription Factor AP-2 / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism*
D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D050656:Transcription Factor AP-2; D008780:Methyltransferases; D011496:Protein Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C516239:SETDB1 protein, mouse