The many faces of plant chromatin: Meeting summary of the 4th European workshop on plant chromatin 2015, Uppsala,Sweden

In June 2015, the fourth European Workshop on Plant Chromatin took place in Uppsala, Sweden, bringing together 80 researchers studying various aspects of plant chromatin and epigenetics. The intricate relationships between plant chromatin dynamics and gene expression change, chromatin organization within the plant cell nucleus, and the impact of chromatin structure on plant development were discussed. Among the main highlights of the meeting were an ever-growing list of newly identified players in chromatin structure establishment and the development of novel tools and approaches to foster our understanding of chromatin-mediated gene regulation, taking into account the context of the plant cell nucleus and its architecture. In this report, we summarize some of the main advances and prospects of plant chromatin research presented at this meeting.     

染色质重塑和高等植物开花时间控制

染色质的结构和组成直接影响转录因子与基因启动子的结合,并最终导致基因的活化或沉默。多年来在酵母和动物等领域的研究已经证实,起关键调节作用的转录因子表达模式的建立和维持需要染色质重塑。外界和细胞内部信号介导的染色质重塑调控基因的表达,并最终调控细胞的分化和生物个体的发育。近几年人们发现高等植物也存在与动物和酵母同源的参与染色质重塑的蛋白质因子。最近的研究结果表明,决定高等植物开花时间关键基因的表达调控就是通过外界信号影响其染色质结构实现的。     

染色质结构之美——染色质可及性

染色质可及性(chromatin accessibility)作为一种衡量染色质结合因子与染色质DNA结合能力高低的染色质属性,是评价染色质结构稳态的重要指标之一,在多种细胞核进程中扮演重要角色,包括基因转录调控以及DNA损伤修复等。该属性的异常调控与多种疾病的发生发展密切相关,包括肿瘤以及神经退行性疾病等。对于该属性探究已经成为生命科学与疾病领域的热点。伴随越来越多的新技术应运而生,例如染色质构象捕获技术、高通量测序技术以及两种技术的结合等。随着技术的进步,多种参与调控染色质可及性的因素被发现和总结,包括核小体占位、组蛋白修饰以及非编码RNA等。多项大规模的染色质组学数据绘制了多种疾病的染色质可及性图谱,为揭示疾病的发生发展与染色质可及性之间的关系提供了数据支持。同时,随着单细胞染色质可及性测序技术的发展,实现了对细胞类型染色质层面的划分,弥补了单纯依赖基因表达划分细胞类型的不足。本文将从染色质的组成与可及性、影响染色质可及性的因素、染色质可及性的检测方法,以及染色质可及性与癌症的关系等方面简要阐述染色质可及性的研究进展。     

动物早期胚胎发育中染色质结构的继承和重编程

染色质开放性和染色质三维高级结构在基因表达和调控中发挥着非常重要的作用,广泛参与分化、发育、肿瘤发生等细胞生理过程,是表观遗传研究的热点领域之一。动物胚胎发育起始于终端分化的卵子受精形成全能性的受精卵。在精卵结合的过程中,染色质开放性和染色质三维高级结构发生了剧烈的变化,经历继承、重编程、重新建立的过程,并指导调控受精卵分化发育最终成为多细胞、多器官组织的新生命个体。本文介绍了近年来研究染色质开放性和染色质三维高级结构的实验分析技术手段,染色质结构在动物早期胚胎发育过程中的变化规律及其在早期胚胎发育中的作用,染色质结构与其他表观遗传信息(甲基化、组蛋白修饰等)关系方面的重要研究进展和存在的科学问题,以期为表观遗传调控早期胚胎发育的研究提供参考。     

A ‘higher order’ of telomere regulation: telomere heterochromatin and telomeric RNAs

Protection of chromosome ends from DNA repair and degradation activities is mediated by specialized protein complexes bound to telomere repeats. Recently, it has become apparent that epigenetic regulation of the telomric chromatin template critically impacts on telomere function and telomere‐length homeostasis from yeast to man. Across all species, telomeric repeats as well as the adjacent subtelomeric regions carry features of repressive chromatin. Disruption of this silent chromatin environment results in loss of telomere‐length control and increased telomere recombination. In turn, progressive telomere loss reduces chromatin compaction at telomeric and subtelomeric domains. The recent discoveries of telomere chromatin regulation during early mammalian development, as well as during nuclear reprogramming, further highlights a central role of telomere chromatin changes in ontogenesis. In addition, telomeres were recently shown to generate long, non‐coding RNAs that remain associated to telomeric chromatin and will provide new insights into the regulation of telomere length and telomere chromatin. In this review, we will discuss the epigenetic regulation of telomeres across species, with special emphasis on mammalian telomeres. We will also discuss the links between epigenetic alterations at mammalian telomeres and telomere‐associated diseases.     

染色质互作相关转录因子的挖掘及功能分析

染色质互作是真核生物基因组组装的基础,并且在调控真核基因细胞特异性表达中发挥重要作用.染色质互作的发生与特定的蛋白质有关,目前已经发现CTCF (CCCTC binding facor,转录阻抑物)和黏连蛋白与染色质互作相关,然而并不清楚是否还有其他蛋白质参与染色质互作.我们将整合高通量染色体构象捕获(Hi-C)和染色质免疫沉淀-测序(ChIP-seq)数据,在GM12878和K562细胞系中挖掘与染色质互作相关的转录因子,并对发现的转录因子做功能分析.我们在频繁发生互作的染色质位点中发现RUNX3、SPI1等转录因子也可能参与染色质互作.另外,通过FP-growth的数据挖掘方法还发现多个转录因子可能协同作用参与染色质互作.研究结果将为染色质互作相关实验的开展提供先验知识.     

Chromatin techniques for plant cells

A large number of recent studies have demonstrated that many important aspects of plant development are regulated by heritable changes in gene expression that do not involve changes in DNA sequence. Rather, these regulatory mechanisms involve modifications of chromatin structure that affect the accessibility of target genes to regulatory factors that can control their expression. The central component of chromatin is the nucleosome, containing the highly conserved histone proteins that are known to be subject to a wide range of post-translational modifications, which act as recognition codes for the binding of chromatin-associated factors. In addition to these histone modifications, DNA methylation can also have a dramatic influence on gene expression. To accommodate the burgeoning interest of the plant science community in the epigenetic control of plant development, a series of methods used routinely in our laboratories have been compiled that can facilitate the characterization of putative chromatin-binding factors at the biochemical, molecular and cellular levels.     

Molecular characterization of the tobacco SET domain protein NtSET1 unravels its role in histone methylation, chromatin binding, and segregation

Plants contain a great number of genes encoding a distinctive class of SET domain proteins which harbor a plant-specific N-terminal part together with a C-terminal part showing highest sequence similarity to the catalytic domain of the yeast CLR4, the human SUV39H1 and G9a histone-methyltransferases (HMTases). Here we show that NtSET1, a representative member of this class from tobacco, methylated both K9 and K27 of histone H3 in vitro. Ectopic expression of NtSET1, by an inducible promoter, increased the amount of dimethylated H3K9 and induced chromosome-segregation defects in tobacco BY2 cells. Deletion analyses show that the HMTase activity, the association with specific chromatin regions and with condensed chromosomes, and the cellular effects largely depended on the C-terminal region including the SET domain of the protein. Nevertheless, the N-terminal part of NtSET1 was capable of targeting the green fluorescent protein to interphase chromatin. Finally, we show that NtSET1 bound LHP1, the Arabidopsis homolog of animal heterochromatin protein 1 (HP1), and that LHP1 co-localized with heterochromatin containing high amounts of dimethylated H3K9, suggesting a role for NtSET1 in heterochromatic function. Taken together, our results provide new insights into the molecular and global chromatin-binding activities of this particular class member of plant SET domain proteins.     

植物三维染色质构型研究进展

染色质在细胞核内的缠绕、折叠及其在细胞核内的空间排布是真核生物染色质构型的主要特征。在经典DNA探针荧光原位杂交显微观察的基础上,基于新一代测序技术的Hi-C及ChIA-PET染色质构型捕获技术已经被广泛应用于动物及植物细胞核染色质构型的研究中,并以新的角度定义了包括:染色体(质)域(chromosome territory)、A/B染色质区室(compartment A/B)、拓扑偶联结构域(topological associated domains,TADs)、染色质环(chromatin loops)等在内的多个更为精细的染色质构型。利用以上两种主流技术,越来越多的植物物种染色质构型特征被鉴定、分析和比较。本文系统分析和总结了近年来以植物细胞为模型的细胞核染色构型领域取得的重要成果,包括各级染色质构型特征的组成、建立机制和主要影响因素等。在此基础上,分析了目前研究植物染色质构型技术的瓶颈和突破性的技术进展,并对后续研究主要关注的问题和研究内容进行了展望,以期为相关领域的研究提供更多的理论参考和依据。     

高等植物中SWI/SNF染色质重塑研究的新进展

染色质重塑是真核生物表观遗传调控的重要方式．通过对染色质物理结构的调节,染色质重塑在高等动植物干细胞的自我更新及分化、器官和个体发育以及肿瘤发生等多种生物学过程中发挥重要作用．近年来,高等动植物染色质重塑方面的研究已经成为表观遗传学研究领域的热点．本综述总结近年来有关高等动植物染色质重塑的重要研究报道,介绍了染色质重塑的结构机制、分析比较了高等动植物染色质重塑复合体的组成及其生物学功能的多样性,并着重综述了高等植物SWI/SNF染色质重塑复合体各组分在调控植物发育与逆境生长等方面的功能,以期为今后植物中染色质重塑的研究提供启示．