Genomic Analysis of Centromere Assembly and Function

着丝粒组装和功能的基因组分析

基本信息

批准号：
10667642
负责人：
BETH A SULLIVAN
金额：
$ 34.79万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10667642
关键词：
Address Architecture Area Artificial Chromosomes Artificial Human Chromosomes Auxins Award Biological Assay Biology Bypass CRISPR/Cas technology Centromere Chimeric Proteins Chromatin Chromosomal Stability Chromosome 17 Chromosome abnormality Chromosomes Classification Competence Congenital Abnormality Coupled DNA DNA Sequence DNA Transposable Elements Defect Dicentric chromosome Disease Elements Engineering Epigenetic Process Exhibits Genome Genome Stability Genome engineering Genomics Genotype Goals Homo sapiens Human Human Chromosomes Human Genome Infertility Kinetochores Knowledge Label Link Location Maintenance Malignant Neoplasms Maps Memory Modeling Molecular Molecular Conformation Outcome Phenotype Population Process Protein Dynamics Proteins Repetitive Sequence Research Role Satellite DNA Series Site Specific qualifier value Stretching Technology Testing Variant Work centromere protein A chromosome movement density experience flexibility functional outcomes genomic locus genomic variation improved innovation monomer nanopore negative affect recruit telomere

Address Architecture Area Artificial Chromosomes Artificial Human Chromosomes Auxins Award Biological Assay Biology Bypass CRISPR/Cas technology Centromere Chimeric Proteins Chromatin Chromosomal Stability Chromosome 17 Chromosome abnormality Chromosomes Classification Competence Congenital Abnormality Coupled DNA DNA Sequence DNA Transposable Elements Defect Dicentric chromosome Disease Elements Engineering Epigenetic Process Exhibits Genome Genome Stability Genome engineering Genomics Genotype Goals Homo sapiens Human Human Chromosomes Human Genome Infertility Kinetochores Knowledge Label Link Location Maintenance Malignant Neoplasms Maps Memory Modeling Molecular Molecular Conformation Outcome Phenotype Population Process Protein Dynamics Proteins Repetitive Sequence Research Role Satellite DNA Series Site Specific qualifier value Stretching Technology Testing Variant Work centromere protein A chromosome movement density experience flexibility functional outcomes genomic locus genomic variation improved innovation monomer nanopore negative affect recruit telomere

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项目摘要

Centromeres are essential to genome inheritance. Abnormal centromere function is associated with birth defects, infertility, and cancer. Human centromeric (CEN) chromatin typically forms on alpha satellite DNA, a 171bp monomeric sequence that is organized into tandem arrays extending for several megabases. The Telomere-to- Telomere (T2T) Consortium that recently produced the first complete human genome assembly revealed that nearly all endogenous human chromosomes contain multiple alpha satellite arrays. We have shown that on some chromosomes, multiple arrays are competent for centromere assembly. Using Homo sapiens chromosome 17 (HSA17) as a model, we demonstrated that centromere location is dictated by genomic variation within alpha satellite DNA. On HSA17 that has three distinct higher order repeat (HOR) unit arrays, when the largest array contains size and sequence variation, centromere assembly shifts to a nearby array. If the centromere forms at a highly variant array, fewer centromere proteins are present and the chromosome experiences instability. We hypothesize that placement/organization of variant HORs within an alpha satellite array influences centromere location and kinetochore assembly. Since most human chromosomes must choose between two (or more) sites at which to build a stable centromere, our work addresses a fundamental gap in the knowledge of basic processes that influence centromere location, competence, and long-term stability. However, we still lack a comprehensive view of the extent of alpha satellite variation within the population and thus the range of functional centromere outcomes. In this competing renewal application, the proposed work builds on our classification of specific alpha satellite variants identified in diverse human populations to assemble stable, de novo centromeres. We will also systematically test centromere competency of long-range alpha satellite organization, coupled with variant content and proximity to mobile elements. Our project goals are to: 1) produce new genomic assemblies of functionally characterized centromeres using targeted long read sequencing approaches, 2) use human artificial chromosome assays to assess competency of different alpha satellite DNA arrangements for de novo centromere formation, 3) explore the molecular basis for variant centromere defects, and 4) use genome engineering to rehabilitate and/or rescue defective variant centromeres. Successful completion of this work will result in major advancement of our basic understanding of genomic variation within large repetitive DNA arrays in humans and its link to specific centromere outcomes and long-term chromosome maintenance and stability.

中心粒对基因组遗传至关重要。异常的中心粒功能与先天缺陷有关，不育和癌症。人的丝粒（CEN）染色质通常在α卫星DNA上形成171bp 单体序列被组织为延伸的几个巨蛋群的串联阵列。端粒到最近产生了第一个完整的人类基因组组装的端粒（T2T）财团表明几乎所有内源性人类染色体都包含多个α卫星阵列。我们已经证明了这一点染色体，多个阵列有能力用于丝粒组装。使用Homo Sapiens染色体17 （HSA17）作为模型，我们证明了Centromere位置由Alpha内的基因组变异决定卫星DNA。在最大数组时，在HSA17上具有三个不同的高阶重复（HOR）单位阵列包含尺寸和序列变化，中心仪组件转移到附近的阵列。如果中心仪形成高度变化的阵列，存在较少的中心粒蛋白，并且染色体体验不稳定。我们假设在Alpha卫星阵列中放置/组织变体HORS会影响Centromere 位置和动力学组装。由于大多数人类染色体必须在两个（或更多）位置之间进行选择在其中建立一个稳定的中心粒，我们的工作解决了基本过程的知识的根本差距这会影响中心位置，能力和长期稳定性。但是，我们仍然缺乏全面的人口内α卫星变化的程度以及功能性丝粒的范围结果。在此相互竞争的续签应用中，拟议的工作以我们对特定alpha的分类为基础卫星变体在各种人群中鉴定出来，以组装稳定的从头centromeres。我们也会远程Alpha卫星组织的系统测试CentRomere的能力，并与变体结合内容和靠近移动元素。我们的项目目标是：1）生产新的基因组组件使用有针对性的长读取测序方法的功能表征着centromeres，2）使用人工评估不同α卫星DNA的能力的染色体测定形成，3）探索变体丝粒缺陷的分子基础，4）使用基因组工程修复和/或救援有缺陷的变体centromeres。成功完成这项工作将导致重大我们对人类重复性DNA阵列中基因组变异的基本理解的发展它与特定的丝粒结局以及长期染色体维持和稳定性的联系。