Epigenetic regulation of Nanog gene in embryonic stem and trophoblast stem cells

The Nanog and Oct‐4 genes are essential for maintaining pluripotency of embryonic stem (ES) cells and early embryos. We previously reported that DNA methylation and chromatin remodeling underlie the cell type‐specific mechanism of Oct‐4 gene expression. In the present study, we found that there is a tissue‐dependent and differentially methylated region (T‐DMR) in the Nanog up‐stream region. The T‐DMR is hypomethylated in ES cells, but is heavily methylated in trophoblast stem (TS) cells and NIH/3T3 cells, in which the Nanog gene is repressed. Furthermore, in vitro methylation of T‐DMR suppressed Nanog promoter activity in reporter assay. Chromatin immunoprecipitation assay revealed that histone H3 and H4 are highly acetylated, and H3 lysine (K) 4 is hypermethylated at the Nanog locus in ES cells. Conversely, histone deacetylation and H3‐K4 demethylation occurred in TS cells. Importantly, in TS cells, hypermethylation of H3‐K9 and ‐K27 is found only at the Nanog locus, not the Oct‐4 locus, indicating that the combination of histone modifications associated with the Nanog gene is distinct from that of the Oct‐4 gene. In conclusion, the Nanog gene is regulated by epigenetic mechanisms involving DNA methylation and histone modifications.

[1]  Kohta Ikegami,et al.  Genome‐wide and locus‐specific DNA hypomethylation in G9a deficient mouse embryonic stem cells , 2007, Genes to cells : devoted to molecular & cellular mechanisms.

[2]  F. Yi,et al.  Repression of Nanog Gene Transcription by Tcf3 Limits Embryonic Stem Cell Self-Renewal , 2006, Molecular and Cellular Biology.

[3]  Fred H. Gage,et al.  Nanog binds to Smad1 and blocks bone morphogenetic protein-induced differentiation of embryonic stem cells , 2006, Proceedings of the National Academy of Sciences.

[4]  B. Turner,et al.  Epigenetic characterization of the early embryo with a chromatin immunoprecipitation protocol applicable to small cell populations , 2006, Nature Genetics.

[5]  Satoshi Tanaka,et al.  DNA Methylation-dependent Epigenetic Regulation of Dimethylarginine Dimethylaminohydrolase 2 Gene in Trophoblast Cell Lineage* , 2006, Journal of Biological Chemistry.

[6]  X. Chen,et al.  The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells , 2006, Nature Genetics.

[7]  H. Cedar,et al.  G9a-mediated irreversible epigenetic inactivation of Oct-3/4 during early embryogenesis , 2006, Nature Cell Biology.

[8]  Y. Matsui,et al.  A histone H3 methyltransferase controls epigenetic events required for meiotic prophase , 2005, Nature.

[9]  A. Clark,et al.  Human embryonic stem cell genes OCT4, NANOG, STELLAR, and GDF3 are expressed in both seminoma and breast carcinoma , 2005, Cancer.

[10]  D. Wheeler,et al.  Orphan Nuclear Receptor GCNF Is Required for the Repression of Pluripotency Genes during Retinoic Acid-Induced Embryonic Stem Cell Differentiation , 2005, Molecular and Cellular Biology.

[11]  K. Robertson.  DNA methylation and human disease , 2005, Nature Reviews Genetics.

[12]  P. Robson,et al.  Transcriptional Regulation of Nanog by OCT4 and SOX2* , 2005, Journal of Biological Chemistry.

[13]  N. Hattori,et al.  Stage-by-Stage Change in DNA Methylation Status of Dnmt1 Locus during Mouse Early Development* , 2005, Journal of Biological Chemistry.

[14]  T. Ushijima,et al.  Detection and interpretation of altered methylation patterns in cancer cells , 2005, Nature Reviews Cancer.

[15]  M. Tada,et al.  Octamer and Sox Elements Are Required for Transcriptional cis Regulation of Nanog Gene Expression , 2005, Molecular and Cellular Biology.

[16]  Ettore Appella,et al.  p53 induces differentiation of mouse embryonic stem cells by suppressing Nanog expression , 2005, Nature Cell Biology.

[17]  L. Olson,et al.  Oct4 expression in adult human stem cells: evidence in support of the stem cell theory of carcinogenesis. , 2004, Carcinogenesis.

[18]  K. Orita,et al.  Demethylating agent, 5-azacytidine, reverses differentiation of embryonic stem cells. , 2004, Biochemical and biophysical research communications.

[19]  Gangning Liang,et al.  Preferential response of cancer cells to zebularine. , 2004, Cancer cell.

[20]  John M. Greally,et al.  Epigenomics: beyond CpG islands , 2004, Nature Reviews Genetics.

[21]  Satoshi Tanaka,et al.  DNA Methylation-mediated Control of Sry Gene Expression in Mouse Gonadal Development* , 2004, Journal of Biological Chemistry.

[22]  Satoshi Tanaka,et al.  Epigenetic Control of Mouse Oct-4 Gene Expression in Embryonic Stem Cells and Trophoblast Stem Cells* , 2004, Journal of Biological Chemistry.

[23]  Y. Pommier,et al.  Inhibition of histone deacetylase increases cytotoxicity to anticancer drugs targeting DNA. , 2003, Cancer research.

[24]  K. Irvine,et al.  Glycosylation regulates Notch signalling , 2003, Nature Reviews Molecular Cell Biology.

[25]  M. Murakami,et al.  The Homeoprotein Nanog Is Required for Maintenance of Pluripotency in Mouse Epiblast and ES Cells , 2003, Cell.

[26]  J. Nichols,et al.  Functional Expression Cloning of Nanog, a Pluripotency Sustaining Factor in Embryonic Stem Cells , 2003, Cell.

[27]  C. Allis,et al.  Trimethylated lysine 9 of histone H3 is a mark for DNA methylation in Neurospora crassa , 2003, Nature Genetics.

[28]  R. Lovell-Badge,et al.  Multipotent cell lineages in early mouse development depend on SOX2 function. , 2003, Genes & development.

[29]  C. Allis,et al.  Induction and maintenance of nonsymmetrical DNA methylation in Neurospora , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[30]  E. Li.  Chromatin modification and epigenetic reprogramming in mammalian development , 2002, Nature Reviews Genetics.

[31]  Satoshi Tanaka,et al.  Epigenetic marks by DNA methylation specific to stem, germ and somatic cells in mice , 2002, Genes to cells : devoted to molecular & cellular mechanisms.

[32]  H. Kato,et al.  G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis. , 2002, Genes & development.

[33]  T. Jenuwein,et al.  The many faces of histone lysine methylation. , 2002, Current opinion in cell biology.

[34]  P. Becker,et al.  Histone acetylation: a switch between repressive and permissive chromatin , 2002, EMBO reports.

[35]  C. Holding,et al.  Human embryonic genes re-expressed in cancer cells , 2001, Oncogene.

[36]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[37]  N. Hattori,et al.  DNA methylation regulates placental lactogen I gene expression. , 2001, Endocrinology.

[38]  S. Ito,et al.  CpG island of rat sphingosine kinase-1 gene: tissue-dependent DNA methylation status and multiple alternative first exons. , 2001, Genomics.

[39]  J. Miyazaki,et al.  Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells , 2000, Nature Genetics.

[40]  A. Bird,et al.  Methylation-Induced Repression— Belts, Braces, and Chromatin , 1999, Cell.

[41]  J. Rossant,et al.  Promotion of trophoblast stem cell proliferation by FGF4. , 1998, Science.

[42]  H. Schöler,et al.  Formation of Pluripotent Stem Cells in the Mammalian Embryo Depends on the POU Transcription Factor Oct4 , 1998, Cell.

[43]  N. Hattori,et al.  Analysis of CpG islands of trophoblast giant cells by restriction landmark genomic scanning. , 1998, Developmental genetics.

[44]  M. Sekiguchi.  Genes to cells: edited by Jun-ichi Tomizawa, Blackwell Science Ltd. Institutional: £218.00 (Europe), £242.00 (Rest of World), US$382.00 (USA and Canada). Individual: £65.00 (Europe), £72.00 (Rest of World), US$114.00 (USA and Canada) ISSN 1356 9597 , 1997 .

[45]  Nihon Hassei Seibutsu Gakkai,et al.  Genes to cells , 1996 .

[46]  K. Okazaki,et al.  Strain difference in establishment of mouse embryonic stem (ES) cell lines. , 1994, The International journal of developmental biology.

[47]  J. Roder,et al.  Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[48]  A. Joyner,et al.  Production of targeted embryonic stem cell clones. , 1993 .

[49]  Donald Metcalf,et al.  Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells , 1988, Nature.

[50]  G. Martin,et al.  Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. , 1981, Proceedings of the National Academy of Sciences of the United States of America.