CONTROL OF VIRUS INDUCED LYSIS

这项研究的长期目标是阐明 噬菌体完成宿主裂解的分子机制。 噬菌体裂解是涉及相互作用的分子的基本现象。 与宿主细胞膜和细菌细胞壁结构。 此外，宿主裂解的过程必须由 以优化噬菌体病毒粒子的生产。由此得出的教训 从噬菌体裂解的研究中学到的知识可能对 对细胞膜结构和动力学及其调控的认识 病毒总体上的扩散，以及对 细菌膜的结构、功能和新陈代谢。噬菌体 系统提供了强大的遗传工具，我们建议将其用于一项研究 Lambda和相关噬菌体以及噬菌体phiX174的裂解系统， 代表了宿主裂解的两种基本策略。我们将进行 编码“霍林”原型的波长S基因的遗传分析， 其功能是允许可溶性内溶酶或溶菌酶通过， 穿过细胞膜。我们的方法使用了积极和消极 用质粒和噬菌体载体筛选Holin功能。变种人 将对噬菌体21的功能同源物进行研究，以确定 21Holin的哪些特征与Lambda Holin有什么共同之处。基因内 将使用抑制和互补来建立点对点 相互作用-霍林序列内的映射。有缺陷的时钟突变体 溶解的时间将被分离和表征。一个试管孔 将开发使用平面脂膜或小分子脂膜的形成试验 单层囊泡。PhiX174 E裂解蛋白之间的相互作用 以及宿主蛋白SlyD，它是人类FKBP的同源物 免疫亲和素，将使用遗传和分子手段进行研究。

The long term objective of this research effort is to elucidate the molecular mechanisms by which bacteriophage accomplish lysis of the host. Phage lysis is a fundamental phenomenon involving molecules which interact with the host cell membrane and with bacterial cell wall structures. Moreover, the process of host lysis must be carefully scheduled by the bacteriophage in order to optimize the production of virions. Thus lessons learned from the study of phage lysis are likely to be important to the understanding of cell membrane structure and dynamics and the regulation of viral proliferation in general, as well as providing insights into the structure, function, and metabolism of the bacterial envelope. Phage systems provide powerful genetic tools, which we propose to use in a study of the lysis systems of lambda and related phage and of the phage phiX174, representing the two fundamental strategies of host lysis. We will conduct a genetic analysis of the lambda S gene, encoding the prototype "holin", which functions to allow passage of the soluble endolysin, or lysozyme, across the cell membrane. Our approach uses positive and negative selections for holin function using plasmid and phage vectors. A mutational study of a functional homolog from phage 21 will be conducted to determine what features of the 21 holin are common with the lambda holin. Intragenic suppression and complementation will be used to establish a point-to-point interaction-map within the holin sequence. Clock mutants defective in the timing of lysis will be isolated and characterized. An in vitro hole forming assay will be developed using planar lipid membranes or small unilamellar vesicles. The interaction between the phiX174 E lysis protein and the host protein SlyD, which is a homolog of the human FKBP immunophilin, will be investigated using genetic and molecular means.