Role of Spo11 and recombination in mouse meiosis

描述（由申请人提供）：我们的目标是了解哺乳动物减数分裂重组和监测它的检查点。减数分裂产生同源染色体之间的物理连接，这对准确分离至关重要。如果重组发生改变，染色体分离就会受到影响，从而导致配子非整倍性。重组的分子机制和防止重组错误的过程还没有很好地理解。小鼠是这些研究的理想选择，因为其与人类相关的分子过程具有广泛的保守性。具体目标是：1。表征小鼠中的交叉稳态。在酵母中，即使当DSB频率降低时，交叉数也保持不变（“交叉稳态”）。同样的过程可以在鼠标中操作。这一假设将通过减少Spo11（编码启动重组的蛋白质的基因）剂量的雄性和雌性动物中重组的细胞学和分子学分析进行检验。2.定义ATM在减数分裂重组中的作用。检查点激酶ATM是修复减数分裂DNA断裂和交叉控制所必需的。将在缺乏ATM的小鼠中确定重组蛋白的行为和重组产物的结构。3.定义Spo11剪接异构体的功能。已经描述了小鼠和人Spo11的多种剪接变体。编码蛋白的功能尚不清楚。这个问题将通过表征携带转基因或表达个体同种型的靶向突变的小鼠来解决。4.探讨TRIP13（PCH2）在监测染色体突触缺陷中的作用。最近在酵母和线虫中的研究表明，染色体突触检查点的存在不同于重组检查点。我们将测试突触检查点蛋白PCH2的小鼠同源物是否在哺乳动物减数分裂中以类似的途径起作用。相关性：卵子或精子中的染色体数目异常会导致发育障碍或自然流产。这些异常通常是由于减数分裂同源重组缺陷引起的染色体分离不当而引起的。这个项目将解决哺乳动物如何控制减数分裂重组和应对时，有问题的基本问题。

DESCRIPTION (provided by applicant): Our goal is to understand both mammalian meiotic recombination and the checkpoints that monitor it. Recombination generates physical connections between homologous chromosomes that are essential for accurate segregation. If recombination is altered, chromosome segregation is affected, and gamete aneuploidy results. The molecular mechanisms of recombination and the processes that protect against recombination errors are not well understood. Mouse is ideal for these studies because of extensive conservation of relevant molecular processes with humans. Specific aims are to: 1. Characterize crossover homeostasis in mouse. In yeast, crossover numbers are maintained even when DSB frequencies are decreased ("crossover homeostasis"). The same process may operate in mouse. This hypothesis will be tested by cytological and molecular analysis of recombination in male and female animals with reduced dosage of Spo11, the gene encoding the protein that initiates recombination. 2. Define the roles of ATM in meiotic recombination. The checkpoint kinase ATM is required for repair of meiotic DMA breaks and for crossover control. Behaviors of recombination proteins and structures of recombination products will be determined in mice lacking ATM. 3. Define the functions of Spo11 splicing isoforms. Multiple splice variants of mouse and human Spo11 have been described. The functions of the encoded proteins are not known. This issue will be addressed through characterization of mice carrying transgenes or targeted mutations that express individual isoforms. 4. To determine the role of TRIP13 (PCH2) in monitoring chromosome synapsis defects. Recent studies in yeast and nematode demonstrated the existence of a chromosome synapsis checkpoint that is distinct from the recombination checkpoint. We will test whether a mouse homolog of the synapsis checkpoint protein PCH2 functions in a similar pathway in mammalian meiosis. Relevance: Abnormal chromosome numbers in eggs or sperm cause developmental disabilities or spontaneous abortion. These abnormalities often arise because of improper separation of chromosomes caused by defects in meiotic homologous recombination. This project will address fundamental questions about how mammals control meiotic recombination and respond when there are problems.