Key Laboratory of Solid Waste Treatment and Resource Recycle, Southwest University of Science and Technology, Mianyang, Sichuan, China.
DNA Cell Biol. 2012 Dec;31(12):1657-64. doi: 10.1089/dna.2012.1708. Epub 2012 Oct 16.
Liquid crystalline phases of DNA and nucleosome core particles can be formed in vitro, indicating the crucial roles of these phases in the maintenance and compaction of genomes in vivo. In the present study, sequential levels of liquid crystalline decompaction were identified in highly purified nuclei of Karenia papilionacea in response to the gradual chelation of divalent counterions by ethylenediaminetetraacetic acid (EDTA); the decompaction was observed using polarizing light microscopy, confocal laser scanning microscopy, and transmission electron microscopy and further confirmed utilizing microcalorimetry. Nested fibrous coils in 150 nm arc-like bands of chromatin were observed in the early stages of chromosomal decompaction. The microcalorimetry spectra of isolated nuclei revealed that the dynamic processes of nuclear decompaction occurred in a nonlinear manner; in addition, an EDTA-sensitive thermal transition between 60°C-70°C, corresponding to a liquid-crystalline-phase transition of chromosomes, was found. The results suggested that nested coils of fibrous chromatin filaments are responsible for the establishment and stabilization of the liquid crystalline and birefringence features of the chromosomes of dinoflagellates. The results also indicated that positively charged divalent counterions play significant roles in modulating liquid crystalline phases to compact the chromosomes of dinoflagellates.
DNA 和核小体核心颗粒的液晶相可以在体外形成,这表明这些相在体内基因组的维持和压缩中起着至关重要的作用。在本研究中,通过逐步螯合二价抗衡离子 EDTA,鉴定了在高度纯化的扇贝多甲藻细胞核中出现的液晶相连续去浓缩的顺序;使用偏光显微镜、共聚焦激光扫描显微镜和透射电子显微镜观察到去浓缩,并用差示扫描量热法进一步确认。在染色体去浓缩的早期阶段,观察到 150nm 弧形带的染色质中嵌套的纤维状线圈。分离核的差示扫描量热法谱表明,核去浓缩的动态过程以非线性方式发生;此外,在 60°C-70°C 之间发现了一个 EDTA 敏感的热转变,对应于染色体的液晶相转变。结果表明,纤维状染色质丝的嵌套线圈负责建立和稳定甲藻染色体的液晶和双折射特征。结果还表明,带正电荷的二价抗衡离子在调节液晶相以压缩甲藻染色体方面起着重要作用。