SGER: Purification of the Yeast Synaptonemal Complex

SGER：酵母联会复合体的纯化

• 批准号: 9911233
• 负责人: David Kaback
• 金额: $ 3万
• 依托单位: Rutgers, The State University of New Jersey-RBHS-New Jersey Med
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9911233&HistoricalAwards=false
• 关键词: SGER; Purification; Yeast; Synaptonemal; Complex

During meiosis reciprocal recombination between homologues appears to be required for proper meiosis I segregation of chromosomes. Interference is the crossover-induced inhibition of additional meiotic recombination and was proposed to be helpful in ensuring that crossovers are evenly distributed over the genome. The mechanism of interference is not known, but the synaptonemal complex (SC) has been proposed to be involved in mediating interference. A better understanding of the molecular mechanism of interference might be gained by investigating the structure of the SC. The only known integral component that is found over the entire length of the SC is the ZIP1 protein. This protein may provide a scaffold for other SC proteins that have not yet been identified. Identification of these components is essential for understanding the mechanisms of interference and other aspects of the meiotic pairing process. This Small Grant for Exploratory Research is aimed at constructing several recombinant ZIP1 constructs that will enable the visualization of the SC as well as the affinity purification of the protein complexes contained in this structure. An existing construct will be modified that is fully functional yet contains green fluorescent protein and affinity tags that will enable the rapid isolation of protein complexes that form with the ZIP1 protein. The identity of any ZIP1 associated proteins will be determined by tandem mass spectroscopy. These studies should enable the identification of the genes corresponding to each protein and provide the groundwork for testing their role in SC complex formation, crossover interference, and other aspects of meiotic recombination and pairing.

在减数分裂过程中，同源物之间的相互重组似乎是适当的减数分裂I染色体分离所必需的。 干扰是交叉引起的抑制额外的减数分裂重组，并被认为有助于确保跨基因均匀分布在基因组上。 干扰机理尚不清楚，但已提出突发型复合物（SC）参与介导干扰。 可以通过研究SC的结构来更好地理解干扰分子机制。 在SC的整个长度上发现的唯一已知的积分分量是Zip1蛋白。 该蛋白可以为尚未鉴定的其他SC蛋白提供脚手架。这些成分的识别对于理解干扰机理和减数分裂配对过程的其他方面至关重要。 这项用于探索性研究的小赠款旨在构建几种重组ZIP1构造，这些ZIP1构造将使SC的可视化以及该结构中包含的蛋白质复合物的亲和力纯化。 现有的构建体将被修改，该构建体功能齐全，但包含绿色荧光蛋白和亲和力标签，该标签将能够快速分离与Zip1蛋白质形成的蛋白质复合物。 任何Zip1相关蛋白的身份将通过串联质谱法确定。 这些研究应能够鉴定与每种蛋白质相对应的基因，并为测试其在SC复合物形成，交叉干扰以及减数分裂重组和配对的其他方面的作用提供基础。