Beauchemin Mathieu, Morse David
Institut de Recherche en Biologie Végétale, Département de Sciences Biologiques, Université de Montréal, Montréal, Québec, H1X 2B2, Canada.
Chromosoma. 2018 Mar;127(1):29-43. doi: 10.1007/s00412-017-0643-8. Epub 2017 Aug 29.
Dinoflagellate chromatin is unique among eukaryotes, as the chromosomes are permanently condensed in a liquid crystal state instead of being packed in nucleosomes. However, how it is organized is still an unsolved mystery, in part due to the lack of a comprehensive catalog of dinoflagellate nuclear proteins. Here, we report the results of CHromatin Enrichment for Proteomics (CHEP) followed by shotgun mass spectrometry sequencing of the chromatin-associated proteins from the dinoflagellate Lingulodinum polyedra. Our analysis identified proteins involved in DNA replication and repair, transcription, and mRNA splicing, and showed a low level of contamination by proteins from other organelles. A limited number of proteins containing DNA-binding domains were found, consistent with the lack of diversity of these proteins in dinoflagellate transcriptomes. However, the number of proteins containing RNA-binding domains was unexpectedly high supporting a potential role for this type of protein in mediating gene expression and chromatin organization. We also identified a number of proteins involved in chromosome condensation and cell cycle progression as well as a single histone protein (H4). Our results provide the first detailed look at the nuclear proteins associated with the unusual chromatin structure of dinoflagellate nuclei and provide important insights into the biochemical basis of its structure and function.
在真核生物中,甲藻染色质是独特的,因为其染色体以液晶态永久浓缩,而非包装在核小体中。然而,它是如何组织的仍是一个未解之谜,部分原因是缺乏甲藻核蛋白的全面目录。在此,我们报告了蛋白质组学染色质富集(CHEP)的结果,随后对来自多面舌形藻的染色质相关蛋白进行了鸟枪法质谱测序。我们的分析鉴定出了参与DNA复制、修复、转录和mRNA剪接的蛋白质,并显示出其他细胞器蛋白质的污染水平较低。发现了数量有限的含有DNA结合结构域的蛋白质,这与甲藻转录组中这类蛋白质的多样性缺乏一致。然而,含有RNA结合结构域的蛋白质数量出乎意料地高,这支持了这类蛋白质在介导基因表达和染色质组织中可能发挥的作用。我们还鉴定出了一些参与染色体浓缩和细胞周期进程的蛋白质以及一种单一的组蛋白(H4)。我们的结果首次详细观察了与甲藻细胞核异常染色质结构相关的核蛋白,并为其结构和功能的生化基础提供了重要见解。