Mechanisms of site-specific DNA recombination

描述：(申请人提供)：由 位点特异性重组酶具有重要的生物学效应。例如, 位点特异性重组负责整合和切除 细菌中噬菌体基因组，细菌染色体的稳定繁殖 原核生物中基因表达的细胞分裂、发育调控和复制 酵母中染色体外成分的数量维持。了解 定点重组的机制将有助于我们更好地理解DNA 发生在原核生物和真核生物中的重排。 此外，该知识可用于在许多情况下操纵反应 有用的方法：在细菌中实现有针对性的基因改变， 酵母、植物、苍蝇和哺乳动物系统。我们使用了特定于FLP站点的 酿酒酵母重组酶作为一种简单的模拟体系 探索DNA-蛋白质相互作用和参与的化学步骤 链断裂和连接反应。这些研究揭示了几个 表征复合反应的全局性机理特征 由整合酶/酪氨酸蛋白家族执行。我们现在将利用 目前可用的分子遗传、生化和结构 进一步解决FLP系统中一些悬而未决的问题的信息。

DESCRIPTION: (provided by applicant): DNA rearrangements mediated by site-specific recombinases have important biological consequences. For example, site-specific recombination is responsible for integration and excision of phage genomes in bacteria, stable propagation of bacterial chromosomes during cell division, developmental control of gene expression in prokaryotes and copy number maintenance of extrachromosomal elements in yeasts. Understanding the mechanism of site-specific recombination will help us better understand DNA rearrangements that occur in living beings, prokaryotic as well as eukaryotic. Furthermore, the knowledge can be used to manipulate the reaction in a number of useful ways: to bring about targeted genetic alterations in bacteria, yeasts, plants, flies and mammalian systems. We have used the Flp site-specific recombinase from Saccharomyces cerevisiae as a simple model system for exploring the DNA-protein interactions and the chemical steps involved in the strand breakage and joining reactions. These studies have unveiled several global mechanistic features that characterize the recombination reactions carried out by the Integrase/Tyrosine family of proteins. We will now utilize the currently available molecular genetic, biochemical and structural information to further tackle some of the unsolved questions in the Flp system.