Molecular Regulation of Mammalian Meiosis

描述(由申请人提供)： 尽管哺乳动物减数分裂具有重要的生物学意义，但它的几个重要特征在很大程度上仍未得到解释。我们和其他人最近的发现表明，这些特征中的许多都由相同的蛋白质PRDM9控制。我们的长期目标是确定这些控制是如何相关的，它们是如何实现的，以及PRDM9如何适应控制减数分裂进展的更大的蛋白质网络。多个PRDM9等位基因在小鼠身上的可获得性将极大地促进这一点，每个等位基因都有自己的调控特异性，在我们拥有的最有用的哺乳动物实验模型中表达出来。PRDM9是一种特殊的蛋白质，在其执行的多种生物学功能中，在其不寻常的结构域结构和在其不同的分子活动中。这项研究将提供的见解从减数分裂延伸到群体遗传学的主要方面，因为热点的位置决定了遗传模式，延伸到识别人类疾病基因的基因图谱研究，以及进化生物学，因为PRDM9的等位基因不亲和性可能导致杂交不育。为了解决相关的实验问题，我们组织了代表不同实验方法的四个项目之间的协调努力：遗传学(项目A)、蛋白质化学(项目B)、计算集成(项目0)、细胞生物学(项目D)和分子生物学(项目A、B、C和D)。我们的动物资源核心将提供小鼠和新的突变品系。一个细胞生物学核心将提供精母细胞的准备并协助芯片实验。计算核心将执行数据管理和分析。我们的长期目标源于以下总体计划目标：a)全面定义PrDM9的S在重组、转录和配子发生中的调控功能；b)通过鉴定和表征PrDM9与基因组DNA、其他减数分裂蛋白和专门的减数分裂结构的相互作用，确定PrDM9在减数分裂调控中的作用；c)通过使用计算技术建立其分子功能的集成模型，确定PrDM9在配子发生、热点激活和转录中的总体作用；以及d)利用PrDM9锌指结构的特殊变异性来阐明基因组调控中普遍存在的ZNFDNA相互作用的复杂性质。该计划项目的结果将显著增加对减数分裂调节机制以及它们参与其他重要生物学过程的理解。

DESCRIPTION (provided by applicant): Despite the biological importance of mammalian meiosis, several of its significant features rema n largely unexplained. Recent discoveries by ourselves and others have shown that many of these features are controlled by the same protein, PRDM9. Our long-term objective is to determine how these controls are related, how they are achieved, and how PRDM9 fits into the larger network of proteins controlling meiosis progression. This will be greatly facilitated by the availability of multiple PRDM9 alleles in mice, each with its own regulatory specificities expressed in the most useful mammalian experimental model that we have. PRDM9 is an exceptional protein in the diversity of biological functions it carries out, in its unusual domain structure, and in its diverse molecular activities. The insights this study will provide extend beyond meiosis to major aspects of population genetics, as the location of hotspots determines patterns of inheritance; to genetic mapping studies identifying human disease genes; and to evolutionary biology, as allelic incompatibilities in PRDM9 can produce hybrid sterility. To address the relevant experimental questions, we have organized a coordinated effort among four projects representing diverse experimental approaches: genetics (Project A), protein chemistry (Project B), computational integration (Project 0), cell biology (Project D), and molecular biology (Projects A, B, C and D). Our Animal Resources Core will provide mice and new mutant strains. A Cell Biology Core will provide preparations of spermatocytes and assist in ChIP experiments. The Computational Core will perform data management and analysis. Our long-term objectives derive from the following overall program goals: a) Comprehensively define PRDM9's regulatory functions in recombination, transcription, and gametogenesis; b) Define the role of PRDM9 in regulation of meiosis by identifying and characterizing its interactions with genomic DNA, other meiotic proteins, and specialized meiotic structures; c) Define the overall role of PRDM9 in gametogenesis, hotspot activation, and transcription by using computational techniques to build an integrated model of its molecular functions; and d) Exploit the exceptional degree of variability in the Zn fingers (ZNF) of PRDM9 to clarify the complex nature of ZNF - DNA interactions, which are so ubiquitous in genomic regulation. Results of the program project will significantly increase the understanding of the mechanisms that regulate meiosis as well as their involvement in other important biological processes.