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首页> 外文期刊>Nucleic Acids Research >IN DIFFERENTIATING MOUSE MYOBLASTS DNA METHYLTRANSFERASE IS POSTTRANSCRIPTIONALLY AND POSTTRANSLATIONALLY REGULATED
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IN DIFFERENTIATING MOUSE MYOBLASTS DNA METHYLTRANSFERASE IS POSTTRANSCRIPTIONALLY AND POSTTRANSLATIONALLY REGULATED

机译：在区分小鼠成肌细胞中，DNA甲基转移酶被后转录和后翻译调控

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摘要

Upon the onset of mouse myoblast differentiation there is a rapid drop in DNA methyltransferase activity followed by a genome wide demethylation [Jost and Jost (1994) J. Biol. Chem. 269, 10040-10043]. Here we show by using specific antibodies directed against DNA methyltransferase that upon differentiation there was a rapid drop in nuclear DNA methyltransferase whilst the internal control histone H1 remained constant. The loss of nuclear methyltransferase was not due to a translocation of the enzyme from the nucleus to the cytoplasm where there was an increase in creatine phosphokinase protein. In vitro run on experiments carried out with growing and differentiating myoblast nuclei showed no difference in the rate of DNA methyltransferase mRNA synthesis. As measured by Northern blot hybridization the relative half life of DNA methyltransferase mRNA in growing and differentiating cells in the presence of Actinomycin D was 5 h and 1 h 30 min respectively, whereas in the same cells the half life of histone H4 mRNA was in both cases 80 min. As measured by a combination of pulse chase experiments with labeled leucine and immunoprecipitation, the relative half-life of DNA methyltransferase in growing and differentiating cells was similar to 18 h and 4 h 30 min respectively.
机译：在小鼠成肌细胞分化开始后，DNA甲基转移酶活性迅速下降，随后是全基因组范围的去甲基化[Jost和Jost（1994）J.Biol.Chem。215：4031-5。化学269，10040-10043]。在这里，我们通过使用针对DNA甲基转移酶的特异性抗体表明，分化后核DNA甲基转移酶迅速下降，而内部对照组蛋白H1保持恒定。核甲基转移酶的丧失不是由于酶从细胞核转移到细胞质，而肌酸磷酸激酶蛋白增加了。用生长和分化成肌细胞核进行的体外实验显示，DNA甲基转移酶mRNA合成速率没有差异。通过Northern印迹杂交测定，在放线菌素D存在下，生长和分化细胞中DNA甲基转移酶mRNA的相对半衰期分别为5 h和1 h 30 min，而在相同细胞中，组蛋白H4 mRNA的半衰期在两个细胞中案件80分钟。通过脉冲追踪实验与标记的亮氨酸和免疫沉淀相结合的测量，DNA甲基转移酶在生长和分化细胞中的相对半衰期分别类似于18 h和4 h 30 min。

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