Jarid2 is a PRC2 component in embryonic stem cells required for multi-lineage differentiation and recruitment of PRC1 and RNA Polymerase II to developmental regulators

Polycomb Repressor Complexes (PRCs) are important regulators of embryogenesis. In embryonic stem (ES) cells many genes that regulate subsequent stages in development are enriched at their promoters for PRC1, PRC2 and Ser 5-phosphorylated RNA Polymerase II (RNAP), and contain domains of 'bivalent' chromatin (enriched for H3K4me3; histone H3 di- or trimethylated at Lys 4 and H3K27me3; histone H3 trimethylated at Lys 27). Loss of individual PRC components in ES cells can lead to gene de-repression and to unscheduled differentiation. Here we show that Jarid2 is a novel subunit of PRC2 that is required for the co-recruitment of PRC1 and RNAP to genes that regulate development in ES cells. Jarid2-deficient ES cells showed reduced H3K4me2/me3 and H3K27me3 marking and PRC1/PRC2 recruitment, and did not efficiently establish Ser 5-phosporylated RNAP at target genes. ES cells lacking Jarid2, in contrast to previously characterized PRC1 and PRC2 mutants, did not inappropriately express PRC2 target genes. Instead, they show a severely compromised capacity for successful differentiation towards neural or mesodermal fates and failed to correctly initiate lineage-specific gene expression in vitro. Collectively, these data indicate that transcriptional priming of bivalent genes in pluripotent ES cells is Jarid2-dependent, and suggests that priming is critical for subsequent multi-lineage differentiation.

[1]  J. Voncken,et al.  Rnf2 (Ring1b) deficiency causes gastrulation arrest and cell cycle inhibition , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[2]  S. Mcconnell,et al.  Gene targeting using a promoterless gene trap vector ("targeted trapping") is an efficient method to mutate a large fraction of genes. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

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

[4]  Megan F. Cole,et al.  Control of Developmental Regulators by Polycomb in Human Embryonic Stem Cells , 2006, Cell.

[5]  A. Spradling,et al.  Searching Chromatin for Stem Cell Identity , 2006, Cell.

[6]  S. Nishikawa,et al.  In vitro differentiation of mouse embryonic stem cells: hematopoietic and vascular cell types. , 2003, Methods in enzymology.

[7]  Kristian Helin,et al.  The Polycomb Group Protein Suz12 Is Required for Embryonic Stem Cell Differentiation , 2007, Molecular and Cellular Biology.

[8]  Stephan Sauer,et al.  Cohesins Functionally Associate with CTCF on Mammalian Chromosome Arms , 2008, Cell.

[9]  Haruhiko Koseki,et al.  Ring1-mediated ubiquitination of H2A restrains poised RNA polymerase II at bivalent genes in mouse ES cells , 2007, Nature Cell Biology.

[10]  Arend Sidow,et al.  Jarid2/Jumonji Coordinates Control of PRC2 Enzymatic Activity and Target Gene Occupancy in Pluripotent Cells , 2009, Cell.

[11]  Austin G Smith,et al.  Defined conditions for neural commitment and differentiation. , 2003, Methods in enzymology.

[12]  T. Magnuson,et al.  The eed mutation disrupts anterior mesoderm production in mice. , 1995, Development.

[13]  J. Zeitlinger,et al.  Polycomb complexes repress developmental regulators in murine embryonic stem cells , 2006, Nature.

[14]  James A. Cuff,et al.  A Bivalent Chromatin Structure Marks Key Developmental Genes in Embryonic Stem Cells , 2006, Cell.

[15]  Debbie L C van den Berg,et al.  An Oct4-Centered Protein Interaction Network in Embryonic Stem Cells , 2010, Cell stem cell.

[16]  V. Orlando,et al.  General transcription factors bind promoters repressed by Polycomb group proteins , 2001, Nature.

[17]  Juri Rappsilber,et al.  JARID2 regulates binding of the Polycomb repressive complex 2 to target genes in ES cells , 2010, Nature.

[18]  S. Orkin,et al.  An Extended Transcriptional Network for Pluripotency of Embryonic Stem Cells , 2008, Cell.

[19]  Terry Magnuson,et al.  Polycomb Repressive Complex 2 Is Dispensable for Maintenance of Embryonic Stem Cell Pluripotency , 2008, Stem cells.

[20]  N. Brockdorff,et al.  Polycomb Repressive Complexes Restrain the Expression of Lineage-Specific Regulators in Embryonic Stem Cells , 2006, Cell cycle.

[21]  Gang Li,et al.  Jarid2 and PRC2, partners in regulating gene expression. , 2010, Genes & development.

[22]  R. Young,et al.  Stem Cells, the Molecular Circuitry of Pluripotency and Nuclear Reprogramming , 2008, Cell.

[23]  Guo-Cheng Yuan,et al.  EZH1 mediates methylation on histone H3 lysine 27 and complements EZH2 in maintaining stem cell identity and executing pluripotency. , 2008, Molecular cell.

[24]  Michael B. Stadler,et al.  Lineage-specific polycomb targets and de novo DNA methylation define restriction and potential of neuronal progenitors. , 2008, Molecular cell.

[25]  Jeroen A. A. Demmers,et al.  Exportin 4 mediates a novel nuclear import pathway for Sox family transcription factors , 2009, The Journal of cell biology.

[26]  R. Young,et al.  A Chromatin Landmark and Transcription Initiation at Most Promoters in Human Cells , 2007, Cell.

[27]  O. Ohara,et al.  Polycomb group proteins Ring1A/B are functionally linked to the core transcriptional regulatory circuitry to maintain ES cell identity , 2008, Development.

[28]  Stephan Sauer,et al.  Chromatin signatures of pluripotent cell lines , 2006, Nature Cell Biology.

[29]  A. Bradley,et al.  Agouti C57BL/6N embryonic stem cells for mouse genetic resources , 2009, Nature Methods.

[30]  S. Orkin,et al.  Jumonji Modulates Polycomb Activity and Self-Renewal versus Differentiation of Stem Cells , 2009, Cell.

[31]  A. Pombo,et al.  Modifications of RNA polymerase II are pivotal in regulating gene expression states , 2009, EMBO reports.

[32]  Megan F. Cole,et al.  Core Transcriptional Regulatory Circuitry in Human Embryonic Stem Cells , 2005, Cell.

[33]  T. Takeuchi,et al.  Roles of jumonji and jumonji family genes in chromatin regulation and development , 2006, Developmental dynamics : an official publication of the American Association of Anatomists.

[34]  V. Kouskoff,et al.  Tracking mesoderm induction and its specification to the hemangioblast during embryonic stem cell differentiation , 2003, Development.

[35]  Kristian Helin,et al.  Suz12 is essential for mouse development and for EZH2 histone methyltransferase activity , 2004, The EMBO journal.

[36]  Kelly J. Morris,et al.  Heterokaryon-Based Reprogramming of Human B Lymphocytes for Pluripotency Requires Oct4 but Not Sox2 , 2008, PLoS genetics.

[37]  Youngsook Lee,et al.  Roles of JUMONJI in mouse embryonic development , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[38]  Y. Schwartz,et al.  Polycomb silencing blocks transcription initiation. , 2004, Molecular cell.

[39]  Yi Zhang,et al.  SUZ12 is required for both the histone methyltransferase activity and the silencing function of the EED-EZH2 complex. , 2004, Molecular cell.