The mechanisms of molecular adaptation to rapidly evolving genes and systems

描述(申请人提供)：最近一项全面的测序工作和随后的严格酵母属进化分析表明，大约123个基因在整个属内快速进化。对快速进化基因的研究与人类健康特别相关，因为人类基因组中的许多快速进化基因都涉及宿主防御和免疫。尽管对人类的进化和遗传研究有后勤上的限制，但这些对萌芽酵母属成员的研究要容易得多。这里提出的实验将使用物理和遗传相互作用图谱来阐明基因组和蛋白质组如何适应酿酒酵母和巴扬酵母中快速进化的基因。在第一个目标中，将在酿酒酵母和贝亚诺链霉菌中确定一组目标三种蛋白质复合体的物理相互作用图，这些复合体都包含一个快速进化的亚基。在第二个目标中，将使用上位性微阵列图谱(E-MAP)技术构建遗传互作图谱，以识别合成的遗传互作 在两组相同的约400个基因敲除菌株之间，导致分析了约160,000个双基因敲除组合。这些结果将与广泛的酿酒酵母E-map数据进行比较，以确定在巴彦链霉菌和酿酒酵母中快速进化的基因在遗传网络连接性方面的差异。综上所述，这些数据将极大地扩大我们对密切相关物种之间快速进化基因的分子适应的理解，并可能为未来研究如何针对微生物的快速进化基因，以减缓或消除耐药病原体的发展提供信息。

DESCRIPTION (provided by applicant): A recent, comprehensive sequencing effort and subsequent evolutionary analysis of the Saccharomyces sensu stricto genus demonstrated that approximately 123 genes are rapidly evolving across the genus. The study of rapidly evolving genes is particularly relevant to human health as many of the rapidly evolving genes in the human genome are involved in host defense and immunity. And although evolutionary and genetic studies on humans have logistical limits, these same studies on members of the budding yeast Saccharomyces genus are much more tractable. The experiments proposed here will use physical and genetic interaction mapping to elucidate how the genome and proteome adapt to accommodate rapidly evolving genes in Saccharomyces cerevisiae and Saccharomyces bayanus. In the first aim, the physical interaction maps of a targeted set of three protein complexes, all containing a rapidly evolving subunit, will be determined in both S. cerevisiae and S. bayanus. In the second aim, a genetic interaction map will be constructed using epistatic mini-array profiling (E-MAP) technology, to identify synthetic genetic interactions between two identical sets of ~400 knockout strains, leading to the analysis of ~160,000 pair wise combinations of double knockouts in S. bayanus. These results will be compared to the extensive set of S. cerevisiae E-MAP data to identify differences in the genetic network connectivity of rapidly evolving genes in S. bayanus and S. cerevisiae. Taken together, these data will substantially expand our understanding of molecular adaptation to rapidly evolving genes among closely related species, and may inform future studies on how to target rapidly evolving genes in microbes so as to slow or eliminate the development of drug-resistant pathogens.