三种方法对草地贪夜蛾基因组转座子的注释

[目的]转座子(Transposable element,TE)是昆虫基因组的重要组成,不同昆虫类群的TE组成、基因组占比及转座活性等基本特征存在巨大差异.本研究旨在探究不同方法对于草地贪夜蛾Spodoptera frugiperda TE的注释效果,并在基因组水平阐明草地贪夜蛾TE的基本特征.[方法]采用3种方法对草地贪夜蛾基因组TE进行预测,包括基于数据库Repbase、ArTEdb进行同源预测,基于重复序列的特性和结构进行从头预测.[结果]ArTEdb方法和从头预测方法鉴定的TE分别占基因组21.48％和27.26％,其中LINE元件无论是拷贝数还是分布密度都最高;其次是DNA元件.2种方法预测的TE分歧率分布峰值约10％,而分歧率＜10％的TE主要是DNA转座子和LINE.比较3种方法的预测结果,Repbase方法灵敏度低,预测的TE远少于其他2种方法.ArTEdb方法能注释出更多的TE,但该方法对于已知超家族鉴定效果不佳.而从头预测注释出的TE数量多,且能划分到不同超家族,甚至能鉴定不包含在Repbase鳞翅目库的TE超家族.[结论]草地贪夜蛾基因组最主要的TE类型是LINE和DNA元件,基因组存在 大量年轻的转座子,草地贪夜蛾在TE家族的组成上与其它鳞翅目物种存在差异.从头预测的方法对草地贪夜蛾基因组TE注释效果较其它2种方法更好.这一研究结果为深入研究转座子的功能及其对草地贪夜蛾基因组多样性奠定了基础.     

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正长达～2m的人类基因组通过多次盘曲折叠,最终被包裹在微小的细胞核中,形成了复杂的基因组三维空间结构,CTCF在基因组三维空间结构的形成和维持中起到了至关重要的作用。在人类基因组上分布着成千上万的CTCF位点,CTCF方向性地结合在这些位点上并且通过环挤出模型形成染色质环,介导远距离DNA元件的相互作用,从而调控发育中基因的时空表达。     

绝缘子调控基因的表达

绝缘子在调控真核基因时空特异表达的过程中起着至关重要的作用.它的主要功能是增强子阻断和异染色质屏障.已经有竞争、阻断和成环等模型描述其增强子阻断功能;而它的异染色质屏障功能主要是通过影响染色质组蛋白的翻译后修饰来实现.已经确定的绝缘子包括果蝇基因组中的染色质特化结构(specialized chromatin structures, scs)和scs、gypsy、鸡珠蛋白β基因座上游的DNaseⅠ高敏感位点cHS4以及小鼠或人Igf2/H19基因座上的印记控制区(imprinting control region, ICR)和DNA甲基化区域(DNA methylated regions, DMR)元件等.许多转录因子参与绝缘子的基因调控作用,例如脊椎动物中的CCCTC结合因子(CCCTC binding factor,CTCF).利用基因组学和生物信息学等方法,还可以在基因组中发现新的绝缘子元件.     

基于迭代自学习的操纵子结构预测

原核生物操纵子结构的准确注释对基因功能和基因调控网络的研究具有重要意义,通过生物信息学方法计算预测是当前基因组操纵子结构注释的最主要来源.当前的预测算法大都需要实验确认的操纵子作为训练集,但实验确认的操纵子数据的缺乏一直成为发展算法的瓶颈.基于对操纵子结构的认识,从基因间距离、转录翻译相关的调控信号以及COG功能注释等特征出发,建立了描述操纵子复杂结构的概率模型,并提出了不依赖于特定物种操纵子数据作为训练集的迭代自学习算法.通过对实验验证的操纵子数据集的测试比较,结果表明算法对于预测操纵子结构非常有效.在不依赖于任何已知操纵子信息的情况下,算法在总体预测水平上超过了目前最好的操纵子预测方法,而且这种自学习的预测算法要优于依赖特定物种进行训练的算法.这些特点使得该算法能够适用于新测序的物种,有别于当前常用的操纵子预测方法.对细菌和古细菌的基因组进行大规模比较分析,进一步提高了对基因组操纵子结构的普遍特征和物种特异性的认识.     

三维基因组测序技术发展

近20年来,研究者们通过大规模DNA测序,在基因组、转录组和表观基因组方面获得了大量的信息,这些信息为人类基因组中的功能元件和相互作用机制等研究提供了许多详细的视图。但明确2 m长的DNA如何组装在10μm左右的细胞核中仍是一个挑战性的工作。人们着眼于多角度探究染色质在狭窄空间里的精准调控表达,因此,三维基因组研究也越来越受到关注,与此相关的技术也在不断发展中。主要介绍以核邻位连接为基础的染色质构象捕获技术(3C)及其衍生技术,如4C、5C、Hi-C及其衍生技术、Ch IA-PET、原位Hi-C,以及DLO Hi-C等,并简单介绍应用这些技术已取得的重要成果以及未来的发展方向。     

蛋白质基因组学：运用蛋白质组技术注释基因组

随着高通量DNA测序技术的飞速发展,越来越多的物种完成了基因组测序．定位编码基因、确定编码基因结构是基因组注释的基本任务,然而以往的基因组注释方法主要依赖于DNA及RNA序列信息．为了更加精确地解读完成测序的基因组,我们需要整合多种类型的组学数据进行基因组注释．近年来,基于串联质谱技术的蛋白质组学已经发展成熟,实现了对蛋白质组的高覆盖,使得利用串联质谱数据进行基因组注释成为可能．串联质谱数据一方面可以对已注释的基因进行表达验证,另一方面还可以校正原注释基因,进而发现新基因,实现对基因组序列的重新注释．这正是当前进展较快的蛋白质基因组学的研究内容．利用该方法系统地注释已完成测序的基因组已成为解读基因组的一个重要补充．本文综述了蛋白质基因组学的主要研究内容和研究方法,并展望了该研究方向未来的发展．     

染色质三维结构重建及其生物学意义

真核生物的基因组在细胞核中以染色质的形式存在,染色质的功能与它的三维结构紧密相关,例如,基因组的复制、转录、调控、DNA突变、长链非编码RNA的传播和胚胎发育等生物功能都是在细胞核的三维空间中完成的.随着染色体构象捕获及其衍生技术与高通量测序技术的结合,产生了大量的染色质交互作用数据.根据这些染色质交互作用数据,研究人员已经提出很多种方法来重建染色质的三维结构.这些方法有助于在不同分辨率下系统地研究染色质的三维结构,为更好地了解染色质的调控功能提供了结构依据.本文总结了近期染色质三维结构建模方法的进展,并探讨了其在研究染色质生物学功能方面的应用.     

人类基因组中的保守非编码元件

比较基因组学的研究发现: 人类基因组中约5%的序列受到选择压力的限制, 但编码序列只占其中很小一部分, 约3.5%是保守、非编码序列。这些保守非编码元件具有重要功能。可能在染色质构型(高级结构)、DNA转录和RNA加工等不同水平参与了基因的表达调控, 与哺乳动物的形态发生和人类疾病相关。文章简要综述了保守非编码元件的识别、功能及验证、起源演化以及与人类疾病的关系。     

转座元件、表观遗传调控与细胞命运决定

转座元件是哺乳动物基因组内含量最多的元素.尽管转座元件的存在对基因组稳定性具有潜在的危险,但它们同时还是潜在的基因调控序列、蛋白质编码序列和染色质结构序列,并参与物种进化过程.因此,基因组中转座元件的有害性和有益性保持着谨慎的平衡,并且这种平衡主要由表观遗传修饰来调控.本文详细介绍了异染色质类型表观遗传修饰如H3K9m...     

核小体及染色质修饰的基因组分布模式和染色质状态

染色质是真核DNA的存在方式,可以通过影响DNA的可及性调节基因转录,其基本单元为核小体,系由约147 bp的DNA缠绕在组蛋白八联体上形成的结构,核小体之间以连接DNA相连．核小体组蛋白上能发生甲基化和乙酰化等化学修饰．核小体位置、DNA的甲基化和组蛋白的修饰等对染色质状态(常染色质或异染色质)及基因组之间的长程相互作用有重要影响．近年,基于高通量测序技术,核小体位置和染色质修饰在多种细胞中的基因组分布已被测定．结果显示,这些标记的分布模式具有位点特异、动态变化、相互偶联和高度复杂的特征．本文详细回顾并评述了核小体位置和染色质修饰的分布模式、对应生物学功能、修饰之间的关联、实验测定技术、染色质状态的计算分析等内容．该工作对于深入认识和理解染色质的表观遗传调节机制有重要意义．     

Discovery of cell-type specific regulatory elements in the human genome using differential chromatin modification analysis

Chromatin modifications have been comprehensively illustrated to play important roles in gene regulation and cell diversity in recent years. Given the rapid accumulation of genome-wide chromatin modification maps across multiple cell types, there is an urgent need for computational methods to analyze multiple maps to reveal combinatorial modification patterns and define functional DNA elements, especially those are specific to cell types or tissues. In this current study, we developed a computational method using differential chromatin modification analysis (dCMA) to identify cell-type-specific genomic regions with distinctive chromatin modifications. We then apply this method to a public data set with modification profiles of nine marks for nine cell types to evaluate its effectiveness. We found cell-type-specific elements unique to each cell type investigated. These unique features show significant cell-type-specific biological relevance and tend to be located within functional regulatory elements. These results demonstrate the power of a differential comparative epigenomic strategy in deciphering the human genome and characterizing cell specificity.     

Advances in computational ChIA-PET data analysis

Genome-wide chromatin interaction analysis has become important for understanding 3D topological structure of a genome as well as for linking distal cis-regulatory elements to their target genes. Compared to the Hi-C method, chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) is unique, in that one can interrogate thousands of chromatin interactions (in a genome) mediated by a specific protein of interest at high resolution and reasonable cost. However, because of the noisy nature of the data, efficient analytical tools have become necessary. Here, we review some new computational methods recently developed by us and compare them with other existing methods. Our intention is to help readers to better understand ChIA-PET results and to guide the users on selection of the most appropriate tools for their own projects.     

Epigenetic regulation of genes during development： A conserved theme from flies to mammals

Eukaryotic genome is organized in form of chromatin within the nucleus. This organization is important for compaction of DNA as well as for the proper expression of the genes. During early embryonic development, genomic packaging receives variety of signals to eventually set up cell type specific expression patterns of genes. This process of regulated chromatinization leads to "cell type specific epigenomes". The expression states attained during differentiation process need to be maintained subsequently throughout the life of the organism. Epigenetie modifications are responsible for chromatin dependent regulatory mechanism and play a key role in maintenance of the expression state-a process referred to as cellular memory. Another key feature in the packaging of the genome is formation of chro- matin domains that are thought to be structural as well as functional units of the higher order chromatin organization. Boundary elements that function to define such domains set the limits of regulatory elements and that of epigenetie modifications. This connection of epige- netic modification, chromatin structure and genome organization has emerged from several studies. Hox genes are among the best studied in this context and have led to the significant understanding of the epigenetic regulation during development. Here we discuss the evolu- tionarily conserved features of epigenetic mechanisms emerged from studies on homeotic gene clusters.     

泛基因组及其在植物功能基因组学研究中的应用

参考基因组是现代功能基因组学的核心框架,以此为基础的现代基因组学技术在过去20年对植物遗传变异发掘、功能基因克隆等研究起了巨大的推动作用.然而,越来越多的研究发现,单一或少数参考基因组不能完整代表和呈现物种或特定群体内的所有基因组变异,因此其在功能基因组学研究中应用存在很大的局限性,甚至会导致错误的结果.泛基因组是指物...     

Methods for identification of epigenetic elements in mammalian long multigenic genome sequences

Epigenetic elements of the genome, i.e. elements that determine stably inherited changes in gene expression without changes in the genomic DNA sequence, are essential tools of genetic regulation in higher eukaryotes. The complete sequencing of the human and other genomes allowed studies to be started on positioning of these elements within long multigenic regions of the genome, which is a prerequisite for a comprehensive functional annotation of genomes. This mini-review considers some recent experimental approaches to the high-throughput identification and mapping of epigenetic elements of mammalian genomes, including the mapping of methylated CpG sites, open and closed chromatin regions, and DNase I hypersensitivity sites.     

Genomic views of chromatin

With the availability of complete genome sequences for a number of organisms, a major challenge has become to understand how chromatin and its epigenetic modifications regulate genome function. High-throughput microarray and sequencing technologies are being combined with biochemical and immunological enrichment methods to obtain genome-scale views of chromatin in a variety of organisms. The data pinpoint novel, genomic elements and expansive chromatin domains, and offer insight into the functions of histone modifications. In parallel, state-of-the-art imaging techniques are being used to investigate higher-order chromatin organization, and are beginning to bridge our understanding of chromatin biology with that of chromosome structure.