Transacting Genes Regulating Recombination Hotspot Activities

调节重组热点活动的基因交易

• 批准号: 8306823
• 负责人: KENNETH PAIGEN
• 金额: $ 20.22万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8306823
• 关键词: Animals; Biological Assay; Biological Process; Chromatin; Chromatin Structure; Chromosome Segregation; Chromosomes; DNA; DNA Sequence; Data; Dependence; Disease; Dose; Ethnic group; Ethylnitrosourea; Eukaryota; Evolution; Failure; Family; Fertility; Frequencies; Gene Conversion; Genes; Genetic; Genetic Crossing Over; Genetic Drift; Genetic Recombination; Genetic Structures; Genetic Variation; Genome; Geography; Grant; Health; Human; Human Genetics; Individual; Individuality; Inherited; Investigation; Knowledge; Learning; Length; Link; Linkage Disequilibrium; Location; Mammals; Maps; Measures; Meiosis; Meiotic Recombination; Molecular; Mus; Mutagenesis; Organism; Pathway interactions; Pattern; Play; Population; Population Biology; Preventive Intervention; Process; Quantitative Trait Loci; Regulation; Regulator Genes; Regulatory Element; Relative (related person); Reproduction; Role; Sampling; Site; Sterility; Stretching; System; Testing; Therapeutic Intervention; Time; Transact; Variant; Vascular Plant; Work; Yeasts; dosage; fungus; genetic regulatory protein; genome wide association study; improved; novel; reproductive; research study; sperm cell; success

DESCRIPTION (provided by applicant): The locations of recombination hotspots in humans and mice and their frequencies of undergoing genetic crossing over determine patterns of linkage disequilibrium and the possibilities for closely linked genes to be co-inherited; both are critical issues in efforts to identify genes important in human health and disease. Our experiments have revealed the existence of a hitherto unknown, macromolecular regulatory system that controls the location and relative activity of mammalian recombination hotspots by activating, suppressing and modulating their activity. Genetic variation in this regulatory system first enabled its discovery and now provides a means of identifying its components and their interactions, an essential step in understanding its mechanisms, as well as its relevance to issues of human genetics, population biology and evolution. The first regulatory protein we identified in this way, PRDM9, enables recombination at a family of mouse hotspots by modifying chromatin structure, allowing formation of the initiating double strand break. Others have simultaneously identified PRDM9 as a regulator of human recombination hotspots. In this project we will identify components of the complementary regulatory system controlling suppression of recombination at specific hotspots, a matter of equal concern in understanding the regulation of recombination. We will identify the genes encoding suppressors of five hotspots on mouse Chr 1 whose activities are regulated by genes that differ between the M. m. domesticus strain C57BL/6J (B6) and the M. m. castaneus strain CAST/EiJ (CAST) by: (a) applying a new, considerably improved, quantitative assay system that measures hotspot activities in sperm DNA samples using NextGen DNA sequencing; (b) using this assay to map and clone the regulatory genes involved; (c) testing their interactions with each other and whether they act in a dose dependent manner, indicating whether they act catalytically or stoichimetrically; and finally, (d) testing whether they control the initiation of recombination or the decision between the alternative recombination pathways leading to crossing over v. non-crossover gene conversions. In separate experiments we will also lay the groundwork for identifying additional regulatory factors with allelic differences between M. m. domesticus (B6) and the M. m. musculus strain PWD. We expect to learn whether each hotspot has its own unique regulatory system or whether there are shared regulatory elements, what these molecules are, whether controls are exerted on the initiation of recombination or the choice between alternate pathways of recombination, and the manner in which any of these genes interact with each other. These data together with the molecular identity of these genes will provide information essential to resolving their mechanism of action. The results will considerably enhance our understanding of one of the most basic of biological processes, genetic recombination.

描述（申请人提供）：人类和小鼠重组热点的位置及其进行遗传交叉的频率决定了连锁不平衡的模式以及紧密连锁的基因共同遗传的可能性；两者都是努力识别对人类健康和疾病重要的基因的关键问题。我们的实验揭示了迄今为止未知的大分子调节系统的存在，该系统通过激活、抑制和调节哺乳动物重组热点的活性来控制其位置和相对活性。这一调控系统中的遗传变异首先促成了它的发现，现在提供了一种识别其组成部分及其相互作用的方法，这是理解其机制及其与人类遗传学、群体生物学和进化问题的相关性的重要一步。我们以这种方式鉴定的第一个调节蛋白 PRDM9，通过修改染色质结构，允许形成起始双链断裂，从而能够在小鼠热点家族中进行重组。其他人同时发现 PRDM9 是人类重组热点的调节因子。 在这个项目中，我们将确定控制特定热点重组抑制的补充调控系统的组成部分，这与理解重组调控同样重要。我们将鉴定编码小鼠 Chr 1 上五个热点抑制因子的基因，这些抑制因子的活性受 M. m. 之间不同的基因调节。家蝇菌株 C57BL/6J (B6) 和 M. m.栗菌株 CAST/EiJ (CAST) 通过： (a) 应用一种新的、显着改进的定量分析系统，使用 NextGen DNA 测序测量精子 DNA 样本中的热点活动； (b) 使用该测定来绘制和克隆所涉及的调控基因； (c) 测试它们之间的相互作用以及它们是否以剂量依赖性方式起作用，表明它们是催化作用还是化学计量作用；最后，（d）测试它们是否控制重组的启动或导致交叉与非交叉基因转换的替代重组途径之间的决定。在单独的实验中，我们还将为识别具有 M. m. 之间等位基因差异的其他调节因子奠定基础。 Domesticus (B6) 和 M. m.肌肉拉伤PWD。 我们期望了解每个热点是否有其独特的调控系统，或者是否存在共享的调控元件，这些分子是什么，是否对重组的启动或重组的替代途径之间的选择进行控制，以及任何重组的方式。这些基因彼此相互作用。这些数据与这些基因的分子特性一起将为解决其作用机制提供必要的信息。 这些结果将大大增强我们对最基本的生物过程之一——基因重组的理解。