中文摘要：噬菌体与宿主之间的相互作用是由噬菌体受体结合蛋白(RBP)介导的。尽管RBP在控制噬菌体活性和宿主范围方面起着重要作用，但其功能的分子规律仍然成谜。本研究开发了一种高通量、位点特异性噬菌体工程方法，系统地剖析了T7噬菌体RBP（1660变体）末端结构域中每个残基的功能作用。得到的丰富数据集可以用于功能谱的跨宿主交叉比较，精确识别功能重要的区域，其中许多区域为以前所未知。替代模式在突变位点及理化性质上显示出宿主特异性差异，揭示了分子对个体宿主的适应性。研究发现了针对抗性宿主的功能获得性变异和消除某些宿主的宿主收缩变异。研究设计高活性T7变异体用于对抗尿路病原体，证明了治疗效用。本研究方法提出了一个描述噬菌体-细菌系统中序列-功能关系的通用框架。
外文摘要：The interaction between a bacteriophage and its host is mediated by the phage's receptor binding protein (RBP). Despite its fundamental role in governing phage activity and host range, molecular rules of RBP function remain a mystery. Here, we systematically dissect the functional role of every residue in the tip domain of T7 phage RBP (1660 variants) by developing a high-throughput, locus-specific, phage engineering method. This rich dataset allowed us to cross compare functional profiles across hosts to precisely identify regions of functional importance, many of which were previously unknown. Substitution patterns showed host-specific differences in position and physicochemical properties of mutations, revealing molecular adaptation to individual hosts. We discovered gain-of-function variants against resistant hosts and host-constricting variants that eliminated certain hosts. To demonstrate therapeutic utility, we engineered highly active T7 variants against a urinary tract pathogen. Our approach presents a generalized framework for characterizing sequence-function relationships in many phage-bacterial systems.
作者：Huss, P；Meger, A；Leander, M；Nishikawa, K；Raman, S
作者单位：Univ Wisconsin
期刊名称：ELIFE
期刊影响因子：7.08
出版年份：2021
出版刊次：10
点击下载：T7噬菌体受体结合域功能谱的深度突变扫描定位