Mínguez A, Franca S, Moreno Díaz de la Espina S
Laboratorio de Biología Celular y Molecular Vegetal, Centro de Investigaciones Biológicas (CSIC), Madrid, Spain.
J Cell Sci. 1994 Oct;107 ( Pt 10):2861-73. doi: 10.1242/jcs.107.10.2861.
Unicellular Dinoflagellates represent the only eukaryotic Phylum lacking histones and nucleosomes. To investigate whether Dinoflagellates do have a nuclear matrix that would modulate the supramolecular organization of their non-nucleosomal DNA and chromosomes, cells of the free-living unarmored Dinoflagellate Amphidinium carterae were encapsulated in agarose microbeads and submitted to sequential extraction with non-ionic detergents, nucleases and 2 M NaCl. Our results demonstrate that this species has a residual nuclear matrix similar to that of vertebrates and higher plants. The cytoskeleton-nuclear matrix complex of A. carterae shows a relatively intricate polypeptide pattern. Immunoblots with different antibodies reveal several intermediate filament types of proteins, one of which is immunologically related to vertebrate lamins, confirming that these proteins are ancestral members of the IF family, which is highly conserved in eukaryotes. A topoisomerase II homologue has also been identified in the nuclear matrix, suggesting that these structures could play a role in organizing the Dinoflagellate DNA in loop domains. Taken together our results demonstrate that the nuclear matrix is an early acquisition of the eukaryotic nucleus, independent of histones and nucleosomes in such a way that the mechanisms controlling the two levels of organization in eukaryotic chromatin would be molecularly and evolutionarily independent.
单细胞甲藻是唯一缺乏组蛋白和核小体的真核生物门。为了研究甲藻是否具有能够调节其非核小体DNA和染色体超分子组织的核基质,将自由生活的无甲单细胞甲藻卡特亚得藻的细胞封装在琼脂糖微珠中,并用非离子洗涤剂、核酸酶和2M氯化钠进行顺序提取。我们的结果表明,该物种具有与脊椎动物和高等植物相似的残余核基质。卡特亚得藻的细胞骨架-核基质复合体显示出相对复杂的多肽模式。用不同抗体进行的免疫印迹揭示了几种中间丝类型的蛋白质,其中一种在免疫上与脊椎动物核纤层蛋白相关,证实这些蛋白质是中间丝家族的祖先成员,在真核生物中高度保守。在核基质中还鉴定出一种拓扑异构酶II同源物,表明这些结构可能在将甲藻DNA组织成环状结构域中发挥作用。综合我们的结果表明,核基质是真核细胞核早期获得的结构,与组蛋白和核小体无关,从而控制真核染色质两个组织水平的机制在分子和进化上是独立的。