Zfp206 Is a Transcription Factor That Controls Pluripotency of Embryonic Stem Cells

Abstract

Zfp206 (ZNF206 in human) encodes a zinc finger-and SCAN domain-containing protein that is highly expressed in pluripotent ESC. Upon differentiation of human and mouse ESC, Zfp206 expression is quickly repressed. Zfp206 was found to be expressed throughout embryogenesis but absent in adult tissues except testis. We have identified a role for Zfp206 in controlling ESC differentiation. ESC engineered to overexpress Zfp206 were found to be resistant to differentiation induced by retinoic acid. In addition, ESC with knocked-down expression of Zfp206 were more sensitive to differentiation by retinoic acid treatment. We found that Zfp206 was able to enhance expression from its own promoter and also activate transcription of the Oct4 and Nanog promoters. Our results show that Zfp206 is an embryonic transcription factor that plays a role in regulating pluripotency of embryonic stem cells.

Zfp206, Oct4, and Sox2 Are Integrated Components of a Transcriptional Regulatory Network in Embryonic Stem Cells

“…Formaldehyde was inactivated by the addition of 125 mM glycine. Sonicated chromatin extracts containing DNA fragments with an average size of 500 bp were immunoprecipitated using 10 g of Zfp206 antibody (13), rabbit IgG (Santa Cruz Biotechnology), or V5 control monoclonal antibodies (Invitrogen) with preblocked protein G-Sepharose beads with overnight incubation at 4°C. The following day, the chromatin-protein-antibody-bead complexes were eluted, and the ChIP DNA was extracted.…”

“…ChIP enrichments were measured by Q-RT-PCR and expressed as fold differences, relative to non-IP (input) chromatin. Positions of the 14 regions (probe numbers [1][2][3][4][5][6][7][8][9][10][11][12][13][14] amplified are indicated as distances (Ϫ6 to 3 kb) from the transcriptional start site (TSS) of the Oct4 gene. B, the transcriptional activity of the Oct4 promoter is dependent on Zfp206-binding sites.…”

“…Zfp206 is another transcription factor that is specifically expressed in ESC (13,14) and is directly regulated by Oct4 and Sox2 (15). Zfp206 was implicated as a pluripotency factor because it was found highly expressed in undifferentiated ESC and the inner cell mass of the preimplantation embryo, but not in differentiated ESC or trophectoderm.…”

“…More recently, thousands of direct target genes regulated by Oct4 and Sox2 have been identified through comprehensive, genome-wide chromatin immunoprecipitation studies (5, 6). Many of the Oct4/Sox2 targets are genes encoding other transcriptional regulators, including several that also have been found to play a role in regulating pluripotency, such as Nanog, Esrrb, Tcf3, Tcl1, Zfp281, Zic3, and Sall4 (3,7,[8][9][10][11][12].Zfp206 is another transcription factor that is specifically expressed in ESC (13,14) and is directly regulated by Oct4 and Sox2 (15). Zfp206 was implicated as a pluripotency factor because it was found highly expressed in undifferentiated ESC and the inner cell mass of the preimplantation embryo, but not in differentiated ESC or trophectoderm.…”

“…Zfp206 was implicated as a pluripotency factor because it was found highly expressed in undifferentiated ESC and the inner cell mass of the preimplantation embryo, but not in differentiated ESC or trophectoderm. Knockdown of Zfp206 expression induces ESC differentiation, whereas its sustained overexpression impedes retinoic acid induced differentiation of ESC, thus establishing that Zfp206 is a regulator of pluripotency (13). Zfp206 encodes a protein that contains 14 zinc fingers, although alternative splice forms contain fewer finger domains.…”