NANOG-dependent function of TET1 and TET2 in establishment of pluripotency.
NANOG-dependent function of TET1 and TET2 in establishment of pluripotency.
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多能状态的分子控制被认为存在于包括同源结构域蛋白Nanog在内的主要转录因子的核心电路中，在体细胞重新编程过程中，Nanog在建立基态多能性方面发挥着至关重要的作用。虽然人们对Nanog的基因组占有率进行了广泛的研究，但对Nanog相关蛋白及其在Nanog介导的重编程过程中的贡献知之甚少。使用增强的纯化技术和严格的计算算法，我们在小鼠ES细胞中鉴定了27个Nanog的高置信度蛋白质相互作用伙伴。其中包括19个未见报道的新的Nanog伴侣，包括Ten-11易位(Tet)家族甲基胞嘧啶羟基酶Tet1。我们证实了Nanog与Tet1的物理结合，并证明了Tet1与Nanog的协同作用提高了重新编程的效率。我们还发现了TET2与Nanog的物理结合和重编程协同作用，并证明了TET2的击倒取消了Nanog与Tet1(Tet1Mut)的催化缺陷突变体的重编程协同作用。这些结果表明，Nanog和Tet1/2蛋白之间的物理相互作用促进了重编程，这种重编程方式依赖于Tet1/2‘S的催化活性。在ES细胞中，Tet1和Nanog共同占据与维持多能性和谱系承诺相关的基因的基因组位置，当Nanog耗尽时，Tet1的结合减少。Nanog和Tet1的共同表达导致在重新编程为天真多能性之前，顶级共同靶标Esrrb和Oct4的表达启动并增加了5hmC水平。我们认为Tet1是由Nanog招募的，以增强关键的重新编程靶基因的子集的表达。这些结果提供了对Nanog重编程机制的洞察，并揭示了5mC羟基酶在建立天真多能性中的新作用。
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 plays an essential role in establishing ground state pluripotency during somatic cell reprogramming. While 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 identified 27 high-confidence protein interaction partners of Nanog in mouse ES cells. These consist of 19 novel partners of Nanog that have not been reported before including the Ten eleven translocation (Tet) family methylcytosine hydroxylase Tet1. We confirmed physical association of Nanog with Tet1, and demonstrated that Tet1, in synergy with Nanog, enhances the efficiency of reprogramming. We also found physical association and reprogramming synergy of Tet2 with Nanog, and demonstrated that knockdown of Tet2 abolishes the reprogramming synergy of Nanog with a catalytically deficient mutant of Tet1 (Tet1Mut). These results indicate that the physical interaction between Nanog and Tet1/2 proteins facilitates reprogramming in a manner that is dependent on Tet1/2's catalytic activity. Tet1 and Nanog co-occupy genomic loci of genes associated with both maintenance of pluripotency and lineage commitment in ES cells, and Tet1 binding is reduced upon Nanog depletion. Co-expression of Nanog and Tet1 results in expression priming of and increased 5hmC levels at top ranked common targets Esrrb and Oct4 before reprogramming to naïve 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 novel role for 5mC hydroxylases in the establishment of naïve pluripotency.
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影响因子：
23.9
作者：
Dawlaty, Meelad M.;Ganz, Kibibi;Powell, Benjamin E.;Hu, Yueh-Chiang;Markoulaki, Styliani;Cheng, Albert W.;Gao, Qing;Kim, Jongpil;Choi, Sang-Woon;Page, David C.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
影响因子：
64.5
作者：
Mitsui, K;Tokuzawa, Y;Yamanaka, S
通讯作者：
Yamanaka, S
影响因子：
64.5
作者：
Marson A;Levine SS;Cole MF;Frampton GM;Brambrink T;Johnstone S;Guenther MG;Johnston WK;Wernig M;Newman J;Calabrese JM;Dennis LM;Volkert TL;Gupta S;Love J;Hannett N;Sharp PA;Bartel DP;Jaenisch R;Young RA
通讯作者：
Young RA
影响因子：
16
作者：
Kallin, Eric M.;Rodriguez-Ubreva, Javier;Christensen, Jesper;Cimmino, Luisa;Aifantis, Iannis;Helin, Kristian;Ballestar, Esteban;Graf, Thomas
通讯作者：
Graf, Thomas
影响因子：
9.8
作者：
Silva, Jose;Barrandon, Ornella;Nichols, Jennifer;Kawaguchi, Jitsutaro;Theunissen, Thorold W.;Smith, Austin
通讯作者：
Smith, Austin