Babinger P, Völkl R, Cakstina I, Maftei A, Schmitt R
Institute for Biochemistry, Genetics and Microbiology, University of Regensburg, 93040 Regensburg, Germany.
Plant Mol Biol. 2007 Feb;63(3):325-36. doi: 10.1007/s11103-006-9091-1. Epub 2006 Oct 11.
DNA methylation plays an important role in the gene-silencing network of higher eukaryotes. We have analyzed the 21.5-kb maintenance methyltransferase (M-MTase) gene, met1, of the multicellular green alga Volvox carteri. The met1 transcript was detected only during the period when DNA replication and cell division are taking place. It encodes a 238 kDa protein containing eight C-terminal activity domains typical of M-MTases, plus upstream DNA-binding domains including the ProDom domain PD003757, which experimental analyses in animal systems have indicated is required for targeting the enzyme to DNA-replication foci. Several insertions of unknown function make Volvox Met1 the largest known member of the Met1/Dnmt1 family. Here we also show that several endogenous transposon families are CpG-methylated in Volvox, which we think causes them to be inactive. This view is supported by the observation that an in vitro CpG-methylated gene introduced into Volvox was maintained in the methylated and silent state over >100 generations. Thus, we believe that Met1 recognizes and perpetuates the in vitro methylation signal, and that the silencing machinery is then able to transduce such a methylation-only signal into a stable heterochromatic (and silent) state.
DNA甲基化在高等真核生物的基因沉默网络中起着重要作用。我们分析了多细胞绿藻团藻的21.5kb维持甲基转移酶(M-MTase)基因met1。met1转录本仅在DNA复制和细胞分裂发生期间被检测到。它编码一种238kDa的蛋白质,该蛋白质包含八个典型的M-MTase C末端活性结构域,以及上游DNA结合结构域,包括ProDom结构域PD003757,动物系统中的实验分析表明,该结构域是将该酶靶向DNA复制位点所必需的。几个功能未知的插入片段使团藻Met1成为Met1/Dnmt1家族中已知最大的成员。在这里,我们还表明,团藻中的几个内源性转座子家族是CpG甲基化的,我们认为这导致它们无活性。这一观点得到以下观察结果的支持:导入团藻的体外CpG甲基化基因在>100代中保持甲基化和沉默状态。因此,我们认为Met1识别并延续体外甲基化信号,然后沉默机制能够将这种仅甲基化的信号转导为稳定的异染色质(沉默)状态。