A permeabilized cell system that assembles filamentous bacteriophage.
A permeabilized cell system has been developed that is capable of assembling filamentous phage only upon addition of exogenous thioredoxin. The in vitro system exhibits the same component requirements seen in vivo: functional thioredoxin, an intact packaging signal in the substrate DNA, and the assembly protein, pIV. This crude in vitro system is insensitive to inhibitors of protein or DNA synthesis, demonstrating that particle assembly uses components that had accumulated before cell permeabilization. The temporal separation of the synthetic period, during which phage proteins and DNA accumulate, from the assembly period enabled us to examine the energy requirement for assembly. We show here that ATP hydrolysis is required for filamentous phage assembly and that the proton motive force is also important.
Feng
J N
JN
Laboratory of Genetics, The Rockefeller University, New York, NY 10021, USA.
Russel
M
M
Model
P
P
D001435:Bacteriophages / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002462:Cell Membrane / Q000821:virology*; D004261:DNA Replication; D004279:DNA, Viral; D018892:Proton-Motive Force; D014779:Virus Replication
D004279:DNA, Viral