Cyclin-dependent kinases regulate epigenetic gene silencing through phosphorylation of EZH2

The Polycomb group (PcG) protein, enhancer of zeste homologue 2 (EZH2), has an essential role in promoting histone H3 lysine 27 trimethylation (H3K27me3) and epigenetic gene silencing. This function of EZH2 is important for cell proliferation and inhibition of cell differentiation, and is implicated in cancer progression. Here, we demonstrate that under physiological conditions, cyclin-dependent kinase 1 (CDK1) and cyclin-dependent kinase 2 (CDK2) phosphorylate EZH2 at Thr 350 in an evolutionarily conserved motif. Phosphorylation of Thr 350 is important for recruitment of EZH2 and maintenance of H3K27me3 levels at EZH2-target loci. Blockage of Thr 350 phosphorylation not only diminishes the global effect of EZH2 on gene silencing, it also mitigates EZH2-mediated cell proliferation and migration. These results demonstrate that CDK-mediated phosphorylation is a key mechanism governing EZH2 function and that there is a link between the cell-cycle machinery and epigenetic gene silencing.

[1]  J. Hsieh,et al.  Down-regulation of Human DAB2IP Gene Expression Mediated by Polycomb Ezh2 Complex and Histone Deacetylase in Prostate Cancer* , 2005, Journal of Biological Chemistry.

[2]  Hengbin Wang,et al.  Role of Histone H3 Lysine 27 Methylation in X Inactivation , 2003, Science.

[3]  G. Hannon,et al.  Ezh2 Orchestrates Gene Expression for the Stepwise Differentiation of Tissue-Specific Stem Cells , 2009, Cell.

[4]  P. Liu,et al.  CDK1 promotes cell proliferation and survival via phosphorylation and inhibition of FOXO1 transcription factor , 2008, Oncogene.

[5]  H. Kovar,et al.  EZH2 is a mediator of EWS/FLI1 driven tumor growth and metastasis blocking endothelial and neuro-ectodermal differentiation , 2009, Proceedings of the National Academy of Sciences.

[6]  S. Dhanasekaran,et al.  The polycomb group protein EZH2 is involved in progression of prostate cancer , 2002, Nature.

[7]  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.

[8]  Kristian Helin,et al.  EZH2 is downstream of the pRB‐E2F pathway, essential for proliferation and amplified in cancer , 2003, The EMBO journal.

[9]  C. Li,et al.  Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Y. Kotake,et al.  pRB family proteins are required for H3K27 trimethylation and Polycomb repression complexes binding to and silencing p16INK4alpha tumor suppressor gene. , 2007, Genes & development.

[11]  D. Tindall,et al.  Androgens repress Bcl-2 expression via activation of the retinoblastoma (RB) protein in prostate cancer cells , 2004, Oncogene.

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

[13]  D. Tindall,et al.  CDK2-Dependent Phosphorylation of FOXO1 as an Apoptotic Response to DNA Damage , 2006, Science.

[14]  D. Reinberg,et al.  Ezh1 and Ezh2 maintain repressive chromatin through different mechanisms. , 2008, Molecular cell.

[15]  Anke Sparmann,et al.  Polycomb silencers control cell fate, development and cancer , 2006, Nature Reviews Cancer.

[16]  J. Labbé,et al.  Identification of major nucleolar proteins as candidate mitotic substrates of cdc2 kinase , 1990, Cell.

[17]  Karima K. Esmail,et al.  A changing paradigm [agile manufacturing] , 1996 .

[18]  S. Elledge,et al.  A quantitative atlas of mitotic phosphorylation , 2008, Proceedings of the National Academy of Sciences.

[19]  Hengbin Wang,et al.  Role of Histone H3 Lysine 27 Methylation in Polycomb-Group Silencing , 2002, Science.

[20]  D. Reinberg,et al.  Histone methyltransferase activity associated with a human multiprotein complex containing the Enhancer of Zeste protein. , 2002, Genes & development.

[21]  V. Pirrotta,et al.  Drosophila Enhancer of Zeste/ESC Complexes Have a Histone H3 Methyltransferase Activity that Marks Chromosomal Polycomb Sites , 2002, Cell.

[22]  H. Piwnica-Worms,et al.  Inactivation of the p34cdc2-cyclin B complex by the human WEE1 tyrosine kinase. , 1992, Science.

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

[24]  Rameen Beroukhim,et al.  An oncogene–tumor suppressor cascade drives metastatic prostate cancer by coordinately activating Ras and nuclear factor-κB , 2010, Nature Medicine.

[25]  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.

[26]  Debashis Ghosh,et al.  EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Brigitte Wild,et al.  Histone Methyltransferase Activity of a Drosophila Polycomb Group Repressor Complex , 2002, Cell.

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

[29]  Vincenzo Pirrotta,et al.  Polycomb silencing mechanisms and the management of genomic programmes , 2007, Nature Reviews Genetics.

[30]  L. Naldini,et al.  Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters , 2005, Nature Biotechnology.

[31]  M. Hung,et al.  Akt-Mediated Phosphorylation of EZH2 Suppresses Methylation of Lysine 27 in Histone H3 , 2005, Science.

[32]  G. Davies,et al.  Knowns and Unknowns , 2003 .

[33]  J. Wagner,et al.  Sleeping Beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[34]  S. Dhanasekaran,et al.  Integrative genomics analysis reveals silencing of beta-adrenergic signaling by polycomb in prostate cancer. , 2007, Cancer cell.

[35]  J. Simon,et al.  Roles of the EZH2 histone methyltransferase in cancer epigenetics. , 2008, Mutation research.

[36]  A. Otte,et al.  Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation , 1999, Nature Genetics.

[37]  S. Varambally,et al.  Genomic Loss of microRNA-101 Leads to Overexpression of Histone Methyltransferase EZH2 in Cancer , 2008, Science.

[38]  John Quackenbush,et al.  Genesis: cluster analysis of microarray data , 2002, Bioinform..

[39]  M. Barbacid,et al.  Cell cycle, CDKs and cancer: a changing paradigm , 2009, Nature Reviews Cancer.

[40]  Robert E. Kingston,et al.  Mechanisms of Polycomb gene silencing: knowns and unknowns , 2009, Nature Reviews Molecular Cell Biology.