Characterization of ion channels involved in the penetration of phage T4 DNA into Escherichia coli cells.
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.
Boulanger
P
P
Laboratoire des Biomembranes, U.A. 1116, Université Paris Sud, Orsay, France.
Letellier
L
L
D000327:Adsorption; D001692:Biological Transport; D002414:Cations, Monovalent; D004279:DNA, Viral / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D004563:Electrochemistry; D004926:Escherichia coli / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000502:physiology* / Q000648:ultrastructure; D007700:Kinetics; D008094:Lithium / Q000378:metabolism; D008564:Membrane Potentials; D011188:Potassium / Q000378:metabolism; D011522:Protons; D012964:Sodium / Q000378:metabolism; D013604:T-Phages / Q000235:genetics / Q000502:physiology*; D013696:Temperature
D002414:Cations, Monovalent; D004279:DNA, Viral; D007473:Ion Channels; D011522:Protons; D008094:Lithium; D004492:Edetic Acid; D012964:Sodium; D011188:Potassium