Institut de Recherche en Biologie Végétale, Département de Sciences Biologiques, Université de Montréal, Montréal, Québec, Canada.
PLoS One. 2012;7(4):e34340. doi: 10.1371/journal.pone.0034340. Epub 2012 Apr 4.
Dinoflagellates typically lack histones and nucleosomes are not observed in DNA spreads. However, recent studies have shown the presence of core histone mRNA sequences scattered among different dinoflagellate species. To date, the presence of all components required for manufacturing and modifying nucleosomes in a single dinoflagellate species has not been confirmed.
Analysis of a Lingulodinium transcriptome obtained by Illumina sequencing of mRNA shows several different copies of each of the four core histones as well as a suite of histone modifying enzymes and histone chaperone proteins. Phylogenetic analysis shows one of each Lingulodinium histone copies belongs to the dinoflagellate clade while the second is more divergent and does not share a common ancestor. All histone mRNAs are in low abundance (roughly 25 times lower than higher plants) and transcript levels do not vary over the cell cycle. We also tested Lingulodinium extracts for histone proteins using immunoblotting and LC-MS/MS, but were unable to confirm histone expression at the protein level.
We show that all core histone sequences are present in the Lingulodinium transcriptome. The conservation of these sequences, even though histone protein accumulation remains below currently detectable levels, strongly suggests dinoflagellates possess histones.
通常情况下,甲藻缺乏组蛋白,且在 DNA 弥散片中观察不到核小体。然而,最近的研究表明,核心组蛋白 mRNA 序列存在于不同的甲藻物种中。迄今为止,尚未在单一甲藻物种中证实存在制造和修饰核小体所需的所有成分。
通过对 Lingulodinium mRNA 进行 Illumina 测序获得的转录组分析表明,每种核心组蛋白都有几个不同的拷贝,以及一系列组蛋白修饰酶和组蛋白伴侣蛋白。系统发育分析表明,Lingulodinium 组蛋白的每个拷贝都属于甲藻分支,而第二个拷贝则更为多样化,没有共同的祖先。所有组蛋白 mRNAs 的丰度都较低(大约比高等植物低 25 倍),且在细胞周期中没有变化。我们还使用免疫印迹和 LC-MS/MS 测试了 Lingulodinium 提取物中的组蛋白蛋白,但未能在蛋白质水平上确认组蛋白的表达。
我们表明,所有核心组蛋白序列都存在于 Lingulodinium 转录组中。尽管组蛋白蛋白的积累仍然低于当前可检测的水平,但这些序列的保守性强烈表明甲藻具有组蛋白。