Analysis of Human Telomeric Structural Variation

人类端粒结构变异分析

基本信息

批准号：
7798586
负责人：
Harold RIETHMAN
金额：
$ 42.01万
依托单位：
WISTAR INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-09-30 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7798586
关键词：
Address Age Aging Alleles Area Base Sequence Biological Biological Assay Biology Blood Cell Line Cell physiology Cells Chromosomal Instability Chromosome Arm Chromosomes Communities Complement Cytogenetics DNA DNA Sequence DNA Sequence Rearrangement DNA Structure Data Databases Development Disease Disease susceptibility Distal Elements Ensure Epigenetic Process Evolution Exhibits Fingerprint Genes Genetic Genetic Materials Genetic Variation Genome Genome Stability Genomic Segment Genotype Goals Heart Diseases Human Human Biology Human Chromosomes Human Genome Incidence Individual Knowledge Length Libraries Malignant Neoplasms Maps Measures Methods Molecular Morphologic artifacts Normal Cell Partner in relationship Phenotype Population Preparation Proteins Qualifying Reading Reagent Regulation Research Resources Role Sampling Site Source Southern Blotting Stretching Structure Testing Validation Variant Work age related assay development base disease phenotype genome sequencing human disease novel public health relevance telomere

项目摘要

DESCRIPTION (provided by applicant): Properly regulated and maintained telomeric DNA is essential for normal genome stability and replication; malfunctioning telomeres are associated with natural aging as well as with a broad spectrum of human diseases including the major killers cancer and heart disease. Although intensive efforts in many labs including our own have led to a reasonably complete reference DNA sequence for most telomeric regions, DNA sequence gaps still exist adjacent to many telomeres. Perhaps more significantly, telomeric and subtelomeric DNA regions exhibit a very high level of structural variation that is likely to impact telomere function but has yet to be characterized in detail. We intend to address these critical gaps in our knowledge of telomeric DNA regions and in our ability to experimentally characterize the role(s) of telomeric DNA in human biology by accomplishing the following three aims: (1) identify and analyze structural variants in distal subtelomeric regions of the human genome, (2) identify and sequence the distal 5 kb of (TTAGGG)n-adjacent subterminal DNA for each telomere allele of multiple unrelated genomes, and (3) develop PCR-based genotyping assays for unique subterminal allelic variant or class of closely-related variants that can be identified. A new resource of clone libraries and associated paired-end reads constructed as part of the Human Structural Variation initiative will be used along with a combination of computational and wet-lab mapping methods to identify and characterize structural variants in distal human subtelomere regions. Terminal fosmids from these libraries will be identified, mapped and used to characterize and sequence (TTAGGG)n-adjacent subterminal DNA. Identification and characterization of telomeric structural variants will close many of the remaining gaps in the human genome sequence, and will in the long term reveal the universe of common germline subterminal allele structures that exist. The global complement of variant subtelomeric alleles in a given individual may have important consequences for expression in gene-rich subtelomeric regions, and could contribute substantially to both natural human phenotypic variation and to disease phenotypes. Creating a database of allele-specific subterminal sequences will help fill a critical gap in our knowledge of telomere structure and its potential impact on length regulation and stability. Appropriately regulated (TTAGGG)n tracts are critical for normal cell function; individual (TTAGGG)n tract lengths in humans are allele-specific and regulated in part by cis-acting subtelomeric elements. The sequence information on subterminal alleles and allelic variants will open new avenues of telomere research and provide novel opportunities for developing PCR-based methods to track subterminal genotypes in populations and measure allele-specific telomere lengths. Public Health Relevance: The tips of human chromosomes have a variable stretch of (TTAGGG)n sequence; this sequence and associated proteins act to ensure proper replication of our genetic material and the stability of our chromosomes. Critically short (TTAGGG)n tracts trigger cells to stop dividing or to self-destruct; if they do not, chromosome instability and perhaps cancer ensues. Cells normally lose (TTAGGG)n slowly with age, suggesting a possible role for these sequences in both aging and the increased cancer incidence with age. DNA sequences adjacent to (TTAGGG)n tracts participate in its regulation and are needed to study biological mechanisms which occur at individual telomeres; however, these sequences are variable among humans and not well defined in the current human genome sequence. This project aims to fill a major gap in our knowledge of the content and variability of (TTAGGG)n-adjacent DNA sequences and to enable understanding of how these sequences help to regulate (TTAGGG)n tract length; hence its completion could have a very significant impact on our our ability to understand of aging and age-related diseases, including cancer.

描述（由申请人提供）：适当调节和维持的端粒DNA对于正常基因组稳定性和复制至关重要；故障端粒与自然衰老以及包括主要杀手癌和心脏病在内的广泛人类疾病有关。尽管在包括我们自己在内的许多实验室中的强烈努力导致了大多数端粒区域的合理完整参考DNA序列，但DNA序列差距仍然存在于许多端粒附近。也许更重要的是，端粒和亚电体DNA区域表现出很高的结构变化，可能会影响端粒功能，但尚未详细表征。我们打算通过实现以下三个目的来解决端粒DNA区域的知识中的这些关键差距，并在实验中表征人类生物学中端粒DNA在人类生物学中的作用的能力：（1）识别和分析人类基因组的远端亚tepelomeric远端基因组中的结构变异，（2）sigens和subles nive and tenter n terminal nive and n tenter n tenter n ternal n tental n tental n tern-tem-tental n terj kb（Ttagg） - ttagg of-tem-tem-tej-te-ttagg of-tem-tenj-------多个无关基因组的等位基因，以及（3）为独特的亚末端等位基因变体或一类紧密相关的变体开发基于PCR的基因分型测定。克隆库和相关的配对末端读取的新资源将与人类结构变化计划的一部分一起构建，并结合计算和湿的LAB映射方法的组合，以识别和表征远端人类亚端子区域中的结构变体。这些文库的末端fosmid将被鉴定，映射和用于表征和序列（TTAGGG）N-近端亚端DNA。端粒结构变体的识别和表征将弥合人类基因组序列中的许多剩余差距，从长远来看，将揭示存在的常见种系亚终端等位基因结构的宇宙。给定个体中变体子层状等位基因的全球补体可能对富基因的亚telomeric区域的表达产生重要影响，并且可以对天然的人类表型变异和疾病表型产生重大贡献。创建等位基因特异性亚终端序列的数据库将有助于填补我们对端粒结构的了解及其对长度调节和稳定性的潜在影响。适当调控（TTAGGG）N区对于正常细胞功能至关重要。人类中的个体（ttaggg）n段长度是等位基因特异性的，部分通过顺式作用的亚电体元素来调节。有关亚终端等位基因和等位基因变体的序列信息将开放端粒研究的新途径，并为开发基于PCR的方法提供了新的机会，以跟踪人群中的亚终端基因型并测量等位基因特异性端粒长度。公共卫生相关性：人类染色体的尖端具有可变的（ttaggg）n序列；该序列和相关的蛋白质起作用，可确保我们的遗传物质和染色体的稳定性正确复制。至关重要的（ttaggg）n区域触发细胞停止分裂或自我毁灭；如果没有，染色体不稳定性和癌症随之而来。细胞通常会随着年龄的增长而缓慢地损失（ttaggg）n，这表明这些序列在衰老和随着年龄的增长率的增加中可能起作用。与（ttaggg）n段相邻的DNA序列参与其调节，需要研究在单个端粒处发生的生物学机制。但是，这些序列在人类中是可变的，在当前人类基因组序列中没有很好地定义。该项目旨在填补我们对（ttaggg）n-Adjacexent DNA序列的内容和可变性的了解的重大差距，并能够理解这些序列如何有助于调节（TTAGGG）N区域长度；因此，它的完成可能会对我们了解包括癌症在内的衰老和年龄相关疾病的能力产生非常重大的影响。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A sequence-ready map of the human chromosome 1q telomere.

人类染色体 1q 端粒的序列就绪图。

DOI：
10.1006/geno.2000.6448
发表时间：
2001
期刊：
Genomics.
影响因子：
0
作者：
Xiang,Z;Morse,E;Hu,XL;Flint,J;Chi,HC;Grady,DL;Moyzis,RK;Riethman,HC
通讯作者：
Riethman,HC

Human subtelomeric copy number variations.

人类亚端粒拷贝数变异。

DOI：
10.1159/000184714
发表时间：
2008
期刊：
Cytogenetic and genome research
影响因子：
1.7
作者：
Riethman,H
通讯作者：
Riethman,H

Physical analysis of the terminal 270 kb of DNA from human chromosome 1q.

对人类 1q 染色体末端 270 kb DNA 进行物理分析。

DOI：
10.1006/geno.1994.1430
发表时间：
1994
期刊：
Genomics
影响因子：
4.4
作者：
Negorev,DG;Macina,RA;Spais,C;Ruthig,LA;Hu,XL;Riethman,HC
通讯作者：
Riethman,HC

Chromosomal bar codes produced by multicolor fluorescence in situ hybridization with multiple YAC clones and whole chromosome painting probes.

DOI：
10.1093/hmg/2.5.505
发表时间：
1993-05
期刊：
Human molecular genetics
影响因子：
3.5
作者：
Christoph Lengauer;Michael R. Speicher;Susanne Popp;Anna Jauch;Masafumi Taniwaki;R. Nagaraja;
通讯作者：
Christoph Lengauer;Michael R. Speicher;Susanne Popp;Anna Jauch;Masafumi Taniwaki;R. Nagaraja;

Dinucleotide repeat polymorphism at the human chromosome 11p telomere (D11S2071).

人类染色体 11p 端粒 (D11S2071) 处的二核苷酸重复多态性。