Center for Plant Molecular Biology (ZMBP), University of Tübingen, Auf der Morgenstelle 32, 72076, Tübingen, Germany.
Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076, Tübingen, Germany.
Genome Biol. 2019 Apr 30;20(1):87. doi: 10.1186/s13059-019-1694-3.
The nuclear envelope not only serves as a physical barrier separating nuclear content from the cytoplasm but also plays critical roles in modulating the three-dimensional organization of genomic DNA. For both plants and animals, the nuclear periphery is a functional compartment enriched with heterochromatin. To date, how plants manage to selectively tether chromatin at the nuclear periphery is unclear.
By conducting dual-color fluorescence in situ hybridization experiments on 2C nuclei, we show that in Arabidopsis thaliana, specific chromatin positioning at the nuclear periphery requires plant lamin-like proteins CROWDED NUCLEI 1 (CRWN1), CRWN4, and DNA methylation in CHG and CHH contexts. With chromosome painting and Hi-C analyses, we show global attenuation of spatial chromatin compartmentalization and chromatin positioning patterns at the nuclear periphery in both the crwn1 and crwn4 mutants. Furthermore, ChIP-seq analysis indicates that CRWN1 directly interacts with chromatin domains localized at the nuclear periphery, which mainly contains non-accessible chromatin.
In summary, we conclude that CRWN1 is a key component of the lamina-chromatin network in plants. It is functionally equivalent to animal lamins, playing critical roles in modulating patterns of chromatin positioning at the nuclear periphery.
核膜不仅作为物理屏障将核内容物与细胞质分隔开,而且在调节基因组 DNA 的三维组织方面也起着关键作用。对于植物和动物来说,核周质富含异染色质,是一个具有功能的隔室。迄今为止,植物如何能够有选择地将染色质固定在核周质尚不清楚。
通过在 2C 核上进行双色荧光原位杂交实验,我们表明在拟南芥中,特定的染色质在核周质的定位需要植物层粘连蛋白样蛋白拥挤核 1(CRWN1)、CRWN4 和 CHG 和 CHH 上下文的 DNA 甲基化。通过染色体作图和 Hi-C 分析,我们表明在 crwn1 和 crwn4 突变体中,空间染色质区室化和核周质的染色质定位模式都受到全局衰减。此外,ChIP-seq 分析表明 CRWN1 与定位于核周质的染色质域直接相互作用,这些染色质域主要包含不可接近的染色质。
总之,我们得出结论,CRWN1 是植物中核膜-染色质网络的关键组成部分。它在功能上与动物 lamin 相当,在调节核周质染色质定位模式方面起着关键作用。