Casjens S, Sampson L, Randall S, Eppler K, Wu H, Petri J B, Schmieger H
Department of Cellular, Viral and Molecular Biology, University of Utah Medical Center, Salt Lake City 84132.
J Mol Biol. 1992 Oct 20;227(4):1086-99. doi: 10.1016/0022-2836(92)90523-m.
Bacteriophage P22 DNA packaging events occur in processive series on concatemeric phage DNA molecules. At the point where such series initiate, the DNA is recognized at a site called pac, and most molecular left ends are generated within six short regions called end sites, which are present in a 120 base-pair region surrounding the pac site. The bacteriophage P22 genes 2 and 3 proteins are required for successful generation of these ends and DNA packaging during progeny virion assembly. Mutants lacking the 162-amino-acid gene 3 protein replicate DNA and assemble functional procapsids. In this report we describe the nucleotide changes and DNA packaging phenotypes of a number of missense mutations of gene 3, which give the phage a higher than normal frequency of generalized transduction. In cells infected by these mutants, more packaging events initiate on the host chromosome than in wild-type infections, so the mutations are thought to affect the specificity of packaging initiation. In addition to having this phenotype, these mutations affect the process of phage DNA packaging in detectable ways. They may: (1) alter the target site specificity for packaging; (2) make target site recognition more promiscuous; (3) affect end site utilization; (4) alter the pac site; and (5) cause apparent random DNA packaging series initiation on phage DNA.
噬菌体P22的DNA包装事件在串联的噬菌体DNA分子上以连续的系列发生。在这些系列开始的位点,DNA在一个称为pac的位点被识别,并且大多数分子的左端在六个称为末端位点的短区域内产生,这些末端位点存在于围绕pac位点的120个碱基对的区域中。噬菌体P22基因2和3的蛋白质是子代病毒体组装过程中成功产生这些末端和进行DNA包装所必需的。缺乏162个氨基酸的基因3蛋白质的突变体能够复制DNA并组装功能性原衣壳。在本报告中,我们描述了基因3的一些错义突变的核苷酸变化和DNA包装表型,这些突变使噬菌体的广义转导频率高于正常水平。在被这些突变体感染的细胞中,与野生型感染相比,在宿主染色体上启动的包装事件更多,因此这些突变被认为会影响包装起始的特异性。除了具有这种表型外,这些突变还以可检测的方式影响噬菌体DNA包装的过程。它们可能:(1)改变包装的靶位点特异性;(2)使靶位点识别更加杂乱;(3)影响末端位点的利用;(4)改变pac位点;(5)导致噬菌体DNA上明显随机的DNA包装系列起始。