Boulanger P, Letellier L
Laboratoire des Biomembranes, U.A. 1116, Université Paris Sud, Orsay, France.
J Biol Chem. 1988 Jul 15;263(20):9767-75.
The hypothesis of a channel-mediated transport of phage DNA into Escherichia coli cytoplasmic membrane has been formulated for a long time. In this paper, we present experimental evidence in favor of this proposal. We have analyzed the kinetics of the K+ efflux induced by T4 phage and ghosts (phage depleted of DNA) using a potassium selective electrode. We show that the K+ efflux is not catalyzed by the K+ transport systems. The Km of K+ efflux is the same for phage and ghosts. The rate of K+ efflux is linearly related to the multiplicity of infection. This suggests that phage and ghosts induce the formation of similar channels and that one channel is induced by one virion. The K+ efflux is associated with an influx of H+ and Na+ or Li+ which compete for entry through the channel. These ion fluxes may be responsible for the cell depolarization. The phage-induced channels allow the passage of DNA. They are only transiently opened, and their closing leads to cellular repolarization. The ghost-induced channels remain open. The insertion and conformation of the channels in the membrane depend on the temperature and their confirmation is voltage-dependent. We give an estimate of their size.
噬菌体DNA通过通道介导转运进入大肠杆菌细胞质膜的假说已经提出很长时间了。在本文中,我们提供了支持这一假说的实验证据。我们使用钾选择性电极分析了T4噬菌体和空斑(不含DNA的噬菌体)诱导的钾离子外流动力学。我们发现钾离子外流不是由钾离子转运系统催化的。噬菌体和空斑的钾离子外流米氏常数相同。钾离子外流速率与感染复数呈线性关系。这表明噬菌体和空斑诱导形成相似的通道,且一个病毒体诱导一个通道。钾离子外流与氢离子、钠离子或锂离子的内流相关,这些离子通过通道竞争进入。这些离子通量可能是细胞去极化的原因。噬菌体诱导的通道允许DNA通过。它们只是短暂开放,其关闭导致细胞复极化。空斑诱导的通道保持开放。通道在膜中的插入和构象取决于温度,且其构象依赖于电压。我们估计了它们的大小。