Bacteriophage T5 DNA ejection under pressure.
The transfer of the bacteriophage genome from the capsid into the host cell is a key step of the infectious process. In bacteriophage T5, DNA ejection can be triggered in vitro by simple binding of the phage to its purified Escherichia coli receptor FhuA. Using electrophoresis and cryo-electron microscopy, we measure the extent of DNA ejection as a function of the external osmotic pressure. In the high pressure range (7-16 atm), the amount of DNA ejected decreases with increasing pressure, as theoretically predicted and observed for lambda and SPP1 bacteriophages. In the low and moderate pressure range (2-7 atm), T5 exhibits an unexpected behavior. Instead of a unique ejected length, multiple populations coexist. Some phages eject their complete genome, whereas others stop at some nonrandom states that do not depend on the applied pressure. We show that contrarily to what is observed for the phages SPP1 and lambda, T5 ejection cannot be explained as resulting from a simple pressure equilibrium between the inside and outside of the capsid. Kinetics parameters and/or structural characteristics of the ejection machinery could play a determinant role in T5 DNA ejection.
Leforestier
A
A
Laboratoire de Physique des Solides, Université Paris-Sud, CNRS UMR 8502, Bât. 510, F-91405 Orsay Cedex, France.
Brasilès
S
S
de Frutos
M
M
Raspaud
E
E
Letellier
L
L
Tavares
P
P
Livolant
F
F
D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001435:Bacteriophages / Q000378:metabolism*; D002213:Capsid; D020285:Cryoelectron Microscopy; D004247:DNA / Q000737:chemistry; D042003:DNA Packaging; D004279:DNA, Viral / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D016678:Genome; D007700:Kinetics; D008954:Models, Biological; D009995:Osmosis; D011312:Pressure; D011485:Protein Binding
D001425:Bacterial Outer Membrane Proteins; D004279:DNA, Viral; D029968:Escherichia coli Proteins; C050900:FhuA protein, E coli; D004247:DNA