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SisterC可检测姐妹染色体相互作用

文章来源:葡京网址;时间:2020-09-24 20:52

本期文章:《自然—方法学》:Online/在线发表

美国马萨诸塞大学医学院Job Dekker研究组取得最新进展。他们发现使用SisterC可检测姐妹染色单体之间以及沿染色单体的染色质相互作用。这一研究成果发表在2020年9月23日的《自然-方法学》上。

他们描述了SisterC,这是一种染色体构象捕获检测法,可区分相同姐妹染色单体之间以及沿着同一姐妹染色单体之间的相互作用。SisterC在S期使用5-溴2'-脱氧尿苷(BrdU)掺入以标记新复制的链,然后进行Hi-C标记,然后通过Hoechst /紫外线处理破坏含5-溴脱氧尿苷的链。在对剩余的完整链进行测序后,这允许根据读取的链将Hi-C产物分为姐妹间和姐妹内相互作用。

他们在有丝分裂的酿酒酵母细胞上进行了SisterC。他们发现着丝粒姐妹染色单体的精确比对。沿臂方向,姐妹染色单体的排列较不精确,姐妹染色体之间,每35kkibase(kb)一个连接。姐妹染色体之间的相互作用发生在黏附蛋白结合位点之间,这些位点之间通常相距5至25 kb。沿着姐妹染色单体,粘着蛋白导致形成高达50 kb的环。SisterC允许研究姐妹染色单体压缩与其在有丝分裂期间的分离之间的复杂相互作用。

附:英文原文

Title: Detecting chromatin interactions between and along sister chromatids with SisterC

Author: Marlies E. Oomen, Adam K. Hedger, Jonathan K. Watts, Job Dekker

Issue&Volume: 2020-09-23

Abstract: Chromosome segregation requires both compaction and disentanglement of sister chromatids. We describe SisterC, a chromosome conformation capture assay that distinguishes interactions between and along identical sister chromatids. SisterC employs 5-bromo-2′-deoxyuridine (BrdU) incorporation during S-phase to label newly replicated strands, followed by Hi-C and then the destruction of 5-bromodeoxyuridine-containing strands via Hoechst/ultraviolet treatment. After sequencing of the remaining intact strands, this allows assignment of Hi-C products as inter- and intra-sister interactions based on the strands that reads are mapped to. We performed SisterC on mitotic Saccharomyces cerevisiae cells. We find precise alignment of sister chromatids at centromeres. Along arms, sister chromatids are less precisely aligned, with inter-sister connections every ~35kilobase (kb). Inter-sister interactions occur between cohesin binding sites that are often offset by 5 to 25kb. Along sister chromatids, cohesin results in the formation of loops of up to 50kb. SisterC allows study of the complex interplay between sister chromatid compaction and their segregation during mitosis.

DOI: 10.1038/s41592-020-0930-9

Source: https://www.nature.com/articles/s41592-020-0930-9

期刊信息

Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:28.467
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex

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