Boulanger P, Letellier L
Laboratoire des Biomembranes, UA 1116 CNRS Université Paris Sud, Orsay, France.
J Biol Chem. 1992 Feb 15;267(5):3168-72.
Phage T5 injects its DNA into Escherichia coli cells in two steps; 8% of the chromosome is first injected, and then there is a pause during which proteins encoded by this DNA fragment are synthesized allowing the remaining DNA to be injected. Using a potassium-selective electrode, we show that the injection of the two DNA fragments is associated with an efflux in two steps, of cytoplasmic potassium. The rate of efflux is linearly related to the number of added phages suggesting that each phage induces the formation of at least one channel in the inner membrane. The first efflux occurs even in depolarized cells suggesting that the insertion and the opening of the channel can take place in the absence of the electrochemical gradient of protons (delta mu H+). The channel is in a closed configuration during the time required for the synthesis of the phage-encoded proteins; this closing and the second efflux are prevented by the depolarization of the cell. The insertion of the channel in the inner membrane requires a fluid membrane. The results obtained suggest that the function of this channel is to translocate phage T5 DNA.
噬菌体T5分两步将其DNA注入大肠杆菌细胞;首先注入8%的染色体,然后有一个停顿期,在此期间由该DNA片段编码的蛋白质被合成,从而使剩余的DNA得以注入。使用钾选择性电极,我们发现这两个DNA片段的注入与细胞质钾的两步外流有关。外流速率与添加的噬菌体数量呈线性关系,这表明每个噬菌体至少在内膜中诱导形成一个通道。即使在去极化细胞中也会发生第一次外流,这表明通道的插入和打开可以在没有质子电化学梯度(δμH+)的情况下发生。在合成噬菌体编码蛋白质所需的时间内,通道处于关闭状态;细胞去极化可防止这种关闭和第二次外流。通道在内膜中的插入需要流动性的膜。所获得的结果表明,该通道的功能是转运噬菌体T5 DNA。