Chow Man H, Yan Kosmo T H, Bennett Michael J, Wong Joseph T Y
Hong Kong University of Science and Technology, Kowloon, Hong Kong.
Eukaryot Cell. 2010 Oct;9(10):1577-87. doi: 10.1128/EC.00026-10. Epub 2010 Apr 16.
DNA can self-assemble in vitro into several liquid crystalline phases at high concentrations. The largest known genomes are encoded by the cholesteric liquid crystalline chromosomes (LCCs) of the dinoflagellates, a diverse group of protists related to the malarial parasites. Very little is known about how the liquid crystalline packaging strategy is employed to organize these genomes, the largest among living eukaryotes-up to 80 times the size of the human genome. Comparative measurements using a semiautomatic polarizing microscope demonstrated that there is a large variation in the birefringence, an optical property of anisotropic materials, of the chromosomes from different dinoflagellate species, despite their apparently similar ultrastructural patterns of bands and arches. There is a large variation in the chromosomal arrangements in the nuclei and individual karyotypes. Our data suggest that both macroscopic and ultrastructural arrangements affect the apparent birefringence of the liquid crystalline chromosomes. Positive correlations are demonstrated for the first time between the level of absolute retardance and both the DNA content and the observed helical pitch measured from transmission electron microscopy (TEM) photomicrographs. Experiments that induced disassembly of the chromosomes revealed multiple orders of organization in the dinoflagellate chromosomes. With the low protein-to-DNA ratio, we propose that a highly regulated use of entropy-driven force must be involved in the assembly of these LCCs. Knowledge of the mechanism of packaging and arranging these largest known DNAs into different shapes and different formats in the nuclei would be of great value in the use of DNA as nanostructural material.
在体外,高浓度的DNA能自组装成几种液晶相。已知最大的基因组由甲藻的胆甾相液晶染色体(LCCs)编码,甲藻是一类与疟原虫相关的多样原生生物。关于这种液晶包装策略如何用于组织这些基因组,人们知之甚少,这些基因组是现存真核生物中最大的——比人类基因组大80倍。使用半自动偏光显微镜进行的比较测量表明,尽管不同甲藻物种的染色体在带和拱形的超微结构模式上看似相似,但它们的双折射(各向异性材料的一种光学特性)存在很大差异。细胞核中的染色体排列和个体核型也存在很大差异。我们的数据表明,宏观和超微结构排列都会影响液晶染色体的表观双折射。首次证明了绝对延迟水平与DNA含量以及从透射电子显微镜(TEM)显微照片测量得到的观察到的螺旋间距之间存在正相关。诱导染色体解组装的实验揭示了甲藻染色体中的多个组织层次。鉴于蛋白质与DNA的比例较低,我们提出这些LCCs的组装过程中必定涉及对熵驱动力的高度调控利用。了解将这些已知最大的DNA包装并排列成细胞核中不同形状和不同形式的机制,对于将DNA用作纳米结构材料具有重要价值。