Zfp281 mediates Nanog autorepression through recruitment of the NuRD complex and inhibits somatic cell reprogramming

Abstract

The homeodomain transcription factor Nanog plays an important role in embryonic stem cell (ESC) self-renewal and is essential for acquiring ground-state pluripotency during reprogramming. Understanding how Nanog is transcriptionally regulated is important for further dissecting mechanisms of ESC pluripotency and somatic cell reprogramming. Here, we report that Nanog is subjected to a negative autoregulatory mechanism, i.e., autorepression, in ESCs, and that such autorepression requires the coordinated action of the Nanog partner and transcriptional repressor Zfp281. Mechanistically, Zfp281 recruits the NuRD repressor complex onto the Nanog locus and maintains its integrity to mediate Nanog autorepression and, functionally, Zfp281-mediated Nanog autorepression presents a roadblock to efficient somatic cell reprogramming. Our results identify a unique transcriptional regulatory mode of Nanog gene expression and shed light into the mechanistic understanding of Nanog function in pluripotency and reprogramming.

NuRD Blocks Reprogramming of Mouse Somatic Cells into Pluripotent Stem Cells

“…5C). This demonstrates that, consistent with a recent report [19], Mbd3 acts as a barrier during the late stage of somatic cell reprogramming by silencing Nanog.…”

“…Thus, depletion of Mbd3/ NuRD upregulates the expression of key iPSC promoting factors, facilitating the generation of pluripotent cells. Apparently, modulatory activity of NuRD constrains differentiation of ESCs thereby directly controlling their immediate differentiation potential [14][15][16]19]. Depletion of other core components of the NuRD complex, such as Mbd2 or Mi2/CHD4, improved iPSC generation to the same extent as knockdown of Mbd3 (unpublished results).…”

“…2H). The observation that downregulation of Mbd3 can substitute for c-Myc or Sox2 in in iPSC generation (Oct4-GFP þ colonies) highlights the essential role of Mbd3 in controlling somatic cell reprogramming and cell fate decision [14,19]. Of note, reprogramming did not occur with Klf4 and c-Myc alone, indicating that depletion of Mbd3/NuRD does not replace Oct4 in iPSC formation.…”

“…In this study, we provided data showing that Mbd3 deficiency facilitates iPSC induction by upregulation of Nanog. NuRD has been shown to be recruited to Nanog by the transcriptional repressor Zfp281 and to dampen expression of genes encoding key pluripotency transcription factors, such as Klf4, Klf5, Zfp42, and Tbx3, in ESCs by opposing transcriptional activators [19]. Wild-type ESCs exhibit bimodal expression of these factors, and this phenotypic variation requires Mbd3.…”

Embryonic stem cell and induced pluripotent stem cell: an epigenetic perspective

“…Such an expression change is associated with LIF-independent self-renewal capacity and deficiency in lineage commitment upon differentiation [40,41]. Moreover, NuRD has been shown to contribute to the autorepression of a set of pluripotency genes, whose expression are subjected to negative autoregulatory feedback control in serum-cultured ESCs [41,42]. In particular, NuRD is recruited by the transcription factor Zfp281 and mediates Nanog autorepression [42].…”