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首页> 外文期刊>Nucleic Acids Research >PHOSPHORYLATION/DEPHOSPHORYLATION OF THE REPRESSOR MDBP-2-H1 SELECTIVELY AFFECTS THE LEVEL OF TRANSCRIPTION FROM A METHYLATED PROMOTER IN VITRO
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PHOSPHORYLATION/DEPHOSPHORYLATION OF THE REPRESSOR MDBP-2-H1 SELECTIVELY AFFECTS THE LEVEL OF TRANSCRIPTION FROM A METHYLATED PROMOTER IN VITRO

机译：阻遏物MDBP-2-H1的磷酸化/去磷酸化选择性影响甲基化启动子的体外转录水平

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摘要

We have previously shown that in vivo estradiol-dependent dephosphorylation of MDBP-2-H1 (a member of the histone H1 family) correlates with the loss of in vitro preferential binding to methylated DNA, To study the effects of the phosphorylation/dephosphorylation of MDBP-2-H1 on the expression of the avian vitellogenin II gene, we optimised an in vitro transcription system using HeLa nuclear extracts, We show that in the absence of the phosphorylated form of MDBP-2-H1 from rooster, methylation of the vitellogenin II promoter does not affect the transcription, Addition of purified MDBP-2-H1 from rooster to the in vitro transcription system inhibits transcription more efficiently from a methylated than an unmethylated DNA template, Dephosphorylation of rooster MDBP-2-H1 by phosphatase treatment or estradiol treatment of rooster lead to the loss of inhibitory activity of the protein when added to the in vitro transcription assays, These findings indicate that the phosphorylation of MDBP-2-H1 is essential for the repression of the transcription, Taken together these results establish the relationship between the dephosphorylation of MDBP-2-H1 caused by estradiol, the down regulation of its binding activity to methylated DNA and the derepression of vitellogenin II transcription.
机译：我们以前已经表明，MDBP-2-H1（组蛋白H1家族成员）的体内雌二醇依赖性去磷酸化与体外优先结合甲基化DNA的丧失有关，以研究MDBP磷酸化/去磷酸化的影响-2-H1在禽卵黄蛋白原II基因的表达上，我们使用HeLa核提取物优化了体外转录系统，我们表明在没有来自公鸡的MDBP-2-H1磷酸化形式的情况下，卵黄蛋白原II的甲基化启动子不影响转录，从公鸡向体外转录系统添加纯化的MDBP-2-H1比未甲基化的DNA模板更有效地抑制从甲基化的转录，通过磷酸酶处理或雌二醇处理使公鸡MDBP-2-H1脱磷酸化当添加到体外转录分析中时，雄性激素导致蛋白质抑制活性的丧失，这些发现表明MDBP-2的磷酸化-H1对于转录的抑制至关重要。综上所述，这些结果建立了雌二醇引起的MDBP-2-H1的去磷酸化，其与甲基化DNA结合活性的下调和卵黄蛋白原II转录的抑制之间的关系。

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