Dunican Donncha S, Pennings Sari, Meehan Richard R
MRC Human Genetics Unit, MRC IGMM, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK.
Centre for Cardiovascular Science, Queen's Medical Research Institute, 47 Little France Crescent, Edinburgh EH16 4TJ, UK.
Biomed Res Int. 2015;2015:740637. doi: 10.1155/2015/740637. Epub 2015 Sep 28.
Eukaryotic genomes are methylated at cytosine bases in the context of CpG dinucleotides, a pattern which is maintained through cell division by the DNA methyltransferase Dnmt1. Dramatic methylation losses are observed in plant and mouse cells lacking Lsh (lymphoid specific helicase), predominantly at repetitive sequences and gene promoters. However, the mechanism by which Lsh contributes to the maintenance of DNA methylation is unknown. Here we show that DNA methylation is lost in Lsh depleted frog and fish embryos, both of which exhibit developmental delay. Additionally, we show that both Lsh and Dnmt1 are associated with chromatin and that Lsh knockdown leads to a decreased Dnmt1-chromatin association. Coimmunoprecipitation experiments reveal that Lsh and Dnmt1 are found in the same protein complex, and pulldowns show this interaction is direct. Our data indicate that Lsh is usually diffuse in the nucleus but can be recruited to heterochromatin in a HP1α-dependent manner. These data together (a) show that the role of Lsh in DNA methylation is conserved in plants, amphibian, fish, and mice and (b) support a model in which Lsh contributes to Dnmt1 binding to chromatin, explaining how its loss can potentially lead to perturbations in DNA methylation maintenance.
真核生物基因组在CpG二核苷酸的背景下,其胞嘧啶碱基会发生甲基化,这种模式通过DNA甲基转移酶Dnmt1在细胞分裂过程中得以维持。在缺乏Lsh(淋巴细胞特异性解旋酶)的植物和小鼠细胞中,观察到显著的甲基化缺失,主要发生在重复序列和基因启动子处。然而,Lsh促进DNA甲基化维持的机制尚不清楚。在此我们表明,在Lsh缺失的青蛙和鱼类胚胎中DNA甲基化会丢失,这两种胚胎均表现出发育延迟。此外,我们表明Lsh和Dnmt1都与染色质相关,并且Lsh基因敲低会导致Dnmt1与染色质的结合减少。免疫共沉淀实验表明,Lsh和Dnmt1存在于同一蛋白质复合物中,下拉实验表明这种相互作用是直接的。我们的数据表明,Lsh通常在细胞核中呈弥散状态,但可以以HP1α依赖的方式被招募到异染色质中。这些数据共同表明:(a)Lsh在DNA甲基化中的作用在植物、两栖动物、鱼类和小鼠中是保守的;(b)支持一种模型,即Lsh有助于Dnmt1与染色质结合,解释了其缺失如何可能导致DNA甲基化维持的扰动。