title: NANOG-dependent function of TET1 and TET2 in establishment of pluripotency.
abstract: Molecular control of the pluripotent state is thought to reside in a core circuitry of master transcription factors including the homeodomain-containing protein NANOG, which has an essential role in establishing ground state pluripotency during somatic cell reprogramming. Whereas the genomic occupancy of NANOG has been extensively investigated, comparatively little is known about NANOG-associated proteins and their contribution to the NANOG-mediated reprogramming process. Using enhanced purification techniques and a stringent computational algorithm, we identify 27 high-confidence protein interaction partners of NANOG in mouse embryonic stem cells. These consist of 19 previously unknown partners of NANOG that have not been reported before, including the ten-eleven translocation (TET) family methylcytosine hydroxylase TET1. We confirm physical association of NANOG with TET1, and demonstrate that TET1, in synergy with NANOG, enhances the efficiency of reprogramming. We also find physical association and reprogramming synergy of TET2 with NANOG, and demonstrate that knockdown of TET2 abolishes the reprogramming synergy of NANOG with a catalytically deficient mutant of TET1. These results indicate that the physical interaction between NANOG and TET1/TET2 proteins facilitates reprogramming in a manner that is dependent on the catalytic activity of TET1/TET2. TET1 and NANOG co-occupy genomic loci of genes associated with both maintenance of pluripotency and lineage commitment in embryonic stem cells, and TET1 binding is reduced upon NANOG depletion. Co-expression of NANOG and TET1 increases 5-hydroxymethylcytosine levels at the top-ranked common target loci Esrrb and Oct4 (also called Pou5f1), resulting in priming of their expression before reprogramming to naive pluripotency. We propose that TET1 is recruited by NANOG to enhance the expression of a subset of key reprogramming target genes. These results provide an insight into the reprogramming mechanism of NANOG and uncover a new role for 5-methylcytosine hydroxylases in the establishment of naive pluripotency.
authors: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK.
authors: Yael
authors: Y
authors: Costa
authors: Junjun
authors: J
authors: Ding
authors: Thorold W
authors: TW
authors: Theunissen
authors: Francesco
authors: F
authors: Faiola
authors: Timothy A
authors: TA
authors: Hore
authors: Pavel V
authors: PV
authors: Shliaha
authors: Miguel
authors: M
authors: Fidalgo
authors: Arven
authors: A
authors: Saunders
authors: Moyra
authors: M
authors: Lawrence
authors: Sabine
authors: S
authors: Dietmann
authors: Satyabrata
authors: S
authors: Das
authors: Dana N
authors: DN
authors: Levasseur
authors: Zhe
authors: Z
authors: Li
authors: Mingjiang
authors: M
authors: Xu
authors: Wolf
authors: W
authors: Reik
authors: José C R
authors: JC
authors: Silva
authors: Jianlong
authors: J
authors: Wang
mesh_terms: D000818:Animals; D065150:Cellular Reprogramming / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D049308:Dioxygenases; D053595:Embryonic Stem Cells; D018507:Gene Expression Regulation, Developmental; D016678:Genome; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D000071317:Nanog Homeobox Protein; D011485:Protein Binding; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D011518:Proto-Oncogene Proteins; C540839:TET1 protein, mouse; D049308:Dioxygenases; C551914:Tet2 protein, mouse
references: Mol Cell. 2012 Oct 26;48(2):266-76
references: 22981865
references: Nat Protoc. 2009;4(4):506-17
references: 19325547
references: Curr Protoc Stem Cell Biol. 2009 Apr;Chapter 4:Unit 4B.1
references: 19382123
references: Nature. 2010 Aug 26;466(7310):1129-33
references: 20639862
references: Curr Protoc Stem Cell Biol. 2009 Mar;Chapter 1:Unit1B.5
references: 19306258
references: Proc Natl Acad Sci U S A. 2008 Apr 29;105(17):6326-31
references: 18436640
references: Cell. 2003 May 30;113(5):631-42
references: 12787504
references: Cell. 2008 Jun 13;133(6):1106-17
references: 18555785
references: Cell. 2009 Aug 21;138(4):722-37
references: 19703398
references: Cell. 2008 Aug 8;134(3):521-33
references: 18692474
references: Nature. 2011 May 19;473(7347):343-8
references: 21490601
references: Cell. 2003 May 30;113(5):643-55
references: 12787505
references: Cell. 2006 Aug 25;126(4):663-76
references: 16904174
references: Cell Res. 2012 Jan;22(1):155-67
references: 22083510
references: Curr Biol. 2011 Jan 11;21(1):65-71
references: 21194951
references: Nature. 2011 May 19;473(7347):389-93
references: 21451524
references: Cell Stem Cell. 2011 Aug 5;9(2):166-75
references: 21816367
references: Cell. 2012 Jun 8;149(6):1368-80
references: 22608086
references: Nature. 2012 Aug 30;488(7413):652-5
references: 22902501
references: Nature. 2009 Dec 3;462(7273):595-601
references: 19898493
references: Science. 2009 May 15;324(5929):930-5
references: 19372391
references: Nat Cell Biol. 2008 Jun;10(6):731-9
references: 18454139
references: PLoS Biol. 2008 Oct 21;6(10):e253
references: 18942890
references: Nature. 2006 Nov 16;444(7117):364-8
references: 17093407
references: Nature. 2011 May 19;473(7347):398-402
references: 21460836
references: J Biol Chem. 2011 Dec 9;286(49):42690-42703
references: 21969378