Henry I, Forlani S, Vaillant S, Muschler J, Choulika A, Nicolas J F
Unité de biologie moléculaire du développement, Institut Pasteur, Paris, France.
C R Acad Sci III. 1999 Dec;322(12):1061-70. doi: 10.1016/s0764-4469(99)00105-5.
The methylation of 5'CpG 3' dinucleotides within genes creates potential targets for protein complexes that bind to methylated DNA sequences and to histone deacetylases (MBD-HDAC). This can lead to transcriptional repression by modification of chromatic. To test the importance of this repression in vivo and to determine when during development these epigenetic controls are placed on genes, two novel genes have been engineered by directed mutagenesis of the CpG-rich LacZ gene that are depleted of (LagZ) or completely lacking (LagoZ) CpG sequences. We report that the expression (transcriptional and translational) of the three genes is indistinguishable in transient assays in cleaving mouse embryos. Therefore, the complete absence of CpG sequences within three kilobases of coding sequence is compatible with its maintenance in the nucleus and with its expression. These molecules can now be used to study the ontogenesis of the CpG-dependent repressive system in intact organisms.
基因内5'CpG 3'二核苷酸的甲基化产生了蛋白质复合物的潜在靶点,这些复合物可与甲基化的DNA序列以及组蛋白脱乙酰酶(MBD-HDAC)结合。这可通过染色质修饰导致转录抑制。为了测试这种抑制在体内的重要性,并确定这些表观遗传控制在发育过程中何时作用于基因,通过对富含CpG的LacZ基因进行定向诱变,构建了两个新基因,它们缺失(LagZ)或完全缺乏(LagoZ)CpG序列。我们报道,在切割小鼠胚胎的瞬时试验中,这三个基因的表达(转录和翻译)没有区别。因此,编码序列三碱基内完全没有CpG序列与其在细胞核中的维持及其表达是相容的。这些分子现在可用于研究完整生物体中CpG依赖性抑制系统的个体发生。