Kapoor Shivali, Zhu Lisha, Froyd Cara, Liu Tao, Rusche Laura N
Department of Biological Sciences, State University of New York at Buffalo, Buffalo, NY 14260;
Department of Biochemistry, State University of New York at Buffalo, Buffalo, NY 14203;
Proc Natl Acad Sci U S A. 2015 Sep 29;112(39):12139-44. doi: 10.1073/pnas.1508749112. Epub 2015 Sep 14.
Point centromeres are specified by a short consensus sequence that seeds kinetochore formation, whereas regional centromeres lack a conserved sequence and instead are epigenetically inherited. Regional centromeres are generally flanked by heterochromatin that ensures high levels of cohesin and promotes faithful chromosome segregation. However, it is not known whether regional centromeres require pericentromeric heterochromatin. In the yeast Candida lusitaniae, we identified a distinct type of regional centromere that lacks pericentromeric heterochromatin. Centromere locations were determined by ChIP-sequencing of two key centromere proteins, Cse4 and Mif2, and are consistent with bioinformatic predictions. The centromeric DNA sequence was unique for each chromosome and spanned 4-4.5 kbp, consistent with regional epigenetically inherited centromeres. However, unlike other regional centromeres, there was no evidence of pericentromeric heterochromatin in C. lusitaniae. In particular, flanking genes were expressed at a similar level to the rest of the genome, and a URA3 reporter inserted adjacent to a centromere was not repressed. In addition, regions flanking the centromeric core were not associated with hypoacetylated histones or a sirtuin deacetylase that generates heterochromatin in other yeast. Interestingly, the centromeric chromatin had a distinct pattern of histone modifications, being enriched for methylated H3K79 and H3R2 but lacking methylation of H3K4, which is found at other regional centromeres. Thus, not all regional centromeres require flanking heterochromatin.
点着丝粒由一个短的共有序列指定,该序列引发动粒形成,而区域着丝粒缺乏保守序列,而是通过表观遗传方式遗传。区域着丝粒通常两侧是异染色质,这确保了高水平的黏连蛋白并促进忠实的染色体分离。然而,尚不清楚区域着丝粒是否需要着丝粒周围的异染色质。在葡萄牙假丝酵母中,我们鉴定出一种独特类型的区域着丝粒,它缺乏着丝粒周围的异染色质。通过对两种关键着丝粒蛋白Cse4和Mif2进行ChIP测序确定了着丝粒位置,并且与生物信息学预测一致。每个染色体的着丝粒DNA序列都是独特的,跨度为4 - 4.5 kbp,这与区域表观遗传遗传着丝粒一致。然而,与其他区域着丝粒不同,在葡萄牙假丝酵母中没有着丝粒周围异染色质的证据。特别是,侧翼基因的表达水平与基因组的其他部分相似,并且插入着丝粒附近的URA3报告基因没有被抑制。此外,着丝粒核心侧翼区域与低乙酰化组蛋白或在其他酵母中产生异染色质的沉默调节蛋白脱乙酰酶无关。有趣的是,着丝粒染色质具有独特的组蛋白修饰模式,富含甲基化的H3K79和H3R2,但缺乏在其他区域着丝粒中发现的H3K4甲基化。因此,并非所有区域着丝粒都需要侧翼异染色质。