Loeb M R
J Virol. 1974 Mar;13(3):631-41. doi: 10.1128/JVI.13.3.631-641.1974.
When Escherichia coli B, labeled by prior growth in (14)C-glucose, are infected with T4 phage there is a rapid release of (14)C-nondialyzable material into the medium. About half of this material is derived from the cell envelope as evidenced by its content of phospholipid and lipopolysaccharide and its buoyant density upon isopycnic ultracentrifugation of 1.19 g/cm(3). It is similar in its gross chemical and physical properties to envelope material released at a lower rate from growing uninfected cells or from cells whose protein synthesis is inhibited by chloramphenicol (22). The rate of release of this envelope material at a multiplicity of infection (MOI) of 10 is greatest in the first minute after infection, and release is completed by 4 min. The rate of its release, as a function of MOI at 2 min after infection, is greatest at low MOI (e.g., MOI 2 and 4); in addition, the release does not continue above MOI 30. The main conclusion derived from the data is that phage, as part of the process of adsorption and injection of DNA, cause an increased release of envelope substance from the cells. With the assumption that all of the envelope material released is derived from the outer envelope, it is estimated that uninfected cells release 20 to 30% of their outer envelope per hour, whereas infected cells release 30% in 2 min at MOI 30. Further, because release does not continue at high MOI, this phenomenon is not considered to be a direct cause of lysis from without. Data are also presented on the amounts of other non-dialyzable (14)C-components released and on the differences in the kinetics of release from chloramphenicol-treated cells compared to phage-infected cells. To avoid the possibility that the release is due to phage lysozyme which is an adventitious "contaminant" of wild-type phage, a phage mutant (T4BeG59s) devoid of this enzyme was used in these experiments.
当用(14)C-葡萄糖预先培养标记的大肠杆菌B被T4噬菌体感染时,有大量不可透析的(14)C物质迅速释放到培养基中。该物质的大约一半来自细胞膜,这可通过其磷脂和脂多糖含量以及在等密度超速离心时1.19 g/cm³的浮力密度得以证明。其总体化学和物理性质与未感染的生长细胞或蛋白质合成受氯霉素抑制的细胞以较低速率释放的细胞膜物质相似(22)。在感染复数(MOI)为10时,这种细胞膜物质在感染后的第一分钟释放速率最大,4分钟时释放完毕。在感染后2分钟,其释放速率作为MOI的函数,在低MOI(例如MOI 2和4)时最大;此外,在MOI 30以上释放不再继续。从这些数据得出的主要结论是,噬菌体在吸附和注入DNA的过程中,会导致细胞释放更多的细胞膜物质。假设释放的所有细胞膜物质都来自外膜,据估计未感染的细胞每小时释放其外膜的20%至30%,而感染的细胞在MOI为30时2分钟内释放30%。此外,由于在高MOI时释放不再继续,所以这种现象不被认为是外部裂解的直接原因。还给出了其他不可透析的(14)C成分释放量的数据,以及与噬菌体感染细胞相比,氯霉素处理细胞释放动力学的差异。为避免释放可能是由于野生型噬菌体偶然“污染”的噬菌体溶菌酶所致,在这些实验中使用了一种不含该酶的噬菌体突变体(T4BeG59s)。