Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute and Harvard School of Public Health, Boston, Massachusetts 02115, USA.
Genome Res. 2011 Feb;21(2):227-36. doi: 10.1101/gr.115519.110. Epub 2010 Dec 22.
Chromatin immunoprecipitation identifies specific interactions between genomic DNA and proteins, advancing our understanding of gene-level and chromosome-level regulation. Based on chromatin immunoprecipitation experiments using validated antibodies, we define the genome-wide distributions of 19 histone modifications, one histone variant, and eight chromatin-associated proteins in Caenorhabditis elegans embryos and L3 larvae. Cluster analysis identified five groups of chromatin marks with shared features: Two groups correlate with gene repression, two with gene activation, and one with the X chromosome. The X chromosome displays numerous unique properties, including enrichment of monomethylated H4K20 and H3K27, which correlate with the different repressive mechanisms that operate in somatic tissues and germ cells, respectively. The data also revealed striking differences in chromatin composition between the autosomes and between chromosome arms and centers. Chromosomes I and III are globally enriched for marks of active genes, consistent with containing more highly expressed genes, compared to chromosomes II, IV, and especially V. Consistent with the absence of cytological heterochromatin and the holocentric nature of C. elegans chromosomes, markers of heterochromatin such as H3K9 methylation are not concentrated at a single region on each chromosome. Instead, H3K9 methylation is enriched on chromosome arms, coincident with zones of elevated meiotic recombination. Active genes in chromosome arms and centers have very similar histone mark distributions, suggesting that active domains in the arms are interspersed with heterochromatin-like structure. These data, which confirm and extend previous studies, allow for in-depth analysis of the organization and deployment of the C. elegans genome during development.
染色质免疫沉淀鉴定了基因组 DNA 与蛋白质之间的特定相互作用,增进了我们对基因水平和染色体水平调控的理解。基于使用经过验证的抗体进行的染色质免疫沉淀实验,我们定义了 19 种组蛋白修饰、1 种组蛋白变体和 8 种染色质相关蛋白在秀丽隐杆线虫胚胎和 L3 幼虫中的全基因组分布。聚类分析确定了 5 组具有共同特征的染色质标记:两组与基因抑制相关,两组与基因激活相关,一组与 X 染色体相关。X 染色体具有许多独特的性质,包括单甲基化 H4K20 和 H3K27 的富集,这与分别在体细胞组织和生殖细胞中起作用的不同抑制机制相关。数据还揭示了常染色体之间以及染色体臂和中心之间染色质组成的惊人差异。染色体 I 和 III 全局富含活性基因标记,与包含更多高表达基因一致,与染色体 II、IV 相比,尤其是 V。与细胞学异染色质的缺失以及秀丽隐杆线虫染色体的全中心性质一致,如 H3K9 甲基化等异染色质标记并未集中在每条染色体的单个区域。相反,H3K9 甲基化在染色体臂上富集,与减数分裂重组升高的区域一致。染色体臂和中心的活性基因具有非常相似的组蛋白标记分布,这表明臂中的活性结构域与异染色质样结构交错。这些数据证实并扩展了以前的研究,使我们能够深入分析秀丽隐杆线虫基因组在发育过程中的组织和部署。