Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
Genes Dev. 2010 Apr 15;24(8):766-82. doi: 10.1101/gad.559610.
To understand whether the spatial organization of the genome reflects the cell's differentiated state, we examined whether genes assume specific subnuclear positions during Caenorhabditis elegans development. Monitoring the radial position of developmentally controlled promoters in embryos and larval tissues, we found that small integrated arrays bearing three different tissue-specific promoters have no preferential position in nuclei of undifferentiated embryos. However, in differentiated cells, they shifted stably toward the nuclear lumen when activated, or to the nuclear envelope when silent. In contrast, large integrated arrays bearing the same promoters became heterochromatic and nuclear envelope-bound in embryos. Tissue-specific activation of promoters in these large arrays in larvae overrode the perinuclear anchorage. For transgenes that carry both active and inactive promoters, the inward shift of the active promoter was dominant. Finally, induction of master regulator HLH-1 prematurely induced internalization of a muscle-specific promoter array in embryos. Fluorescence in situ hybridization confirmed analogous results for the endogenous endoderm-determining gene pha-4. We propose that, in differentiated cells, subnuclear organization arises from the selective positioning of active and inactive developmentally regulated promoters. We characterize two forces that lead to tissue-specific subnuclear organization of the worm genome: large repeat-induced heterochromatin, which associates with the nuclear envelope like repressed genes in differentiated cells, and tissue-specific promoters that shift inward in a dominant fashion over silent promoters, when they are activated.
为了理解基因组的空间组织是否反映了细胞的分化状态,我们研究了在秀丽隐杆线虫的发育过程中,基因是否会在核内呈现特定的亚核位置。通过监测胚胎和幼虫组织中发育调控启动子的径向位置,我们发现,带有三个不同组织特异性启动子的小型整合阵列在未分化胚胎的核内没有优先位置。然而,在分化的细胞中,当它们被激活时,这些启动子会稳定地向核腔转移,或者在沉默时向核膜转移。相比之下,带有相同启动子的大型整合阵列在胚胎中变成异染色质并与核膜结合。在幼虫中,这些大型阵列中相同启动子的组织特异性激活克服了核周锚定。对于携带活性和非活性启动子的转基因,活性启动子的内向移位占主导地位。最后,HLH-1 主调控因子的诱导过早地诱导了胚胎中肌肉特异性启动子阵列的内化。荧光原位杂交证实了内胚层决定基因 pha-4 的类似结果。我们提出,在分化的细胞中,亚核组织的产生源于活性和非活性发育调控启动子的选择性定位。我们描述了导致线虫基因组组织特异性亚核组织的两种力量:大的重复诱导异染色质,其与核膜的结合方式类似于分化细胞中被抑制的基因;组织特异性启动子在被激活时以显性方式向内移位,超过沉默的启动子。