Analysis of the coliphage T5 DNA ejection process with free and liposome-associated TonA protein.
Outer membrane protein TonA, the receptor for coliphage T5, has been partially purified and incorporated into the phospholipid bilayer of liposomes. Adsorption of the phage to its receptor in either a free or liposome-associated form is fast and sufficient to trigger the ejection of encapsidated DNA. In both in vitro systems the exit of DNA from the phage capsid is a very slow process. Ejected DNA can partially accumulate inside the liposome aqueous compartment, but the transfer from the phage head to the liposome internal space is never complete, perhaps because the liposome volume is too small. The presence of polyamines or divalent cations (magnesium) or both in the incubation medium diminished the extent of DNA ejection, possibly by stabilizing DNA inside the head. DNA movement was slowed as the temperature was decreased from 37 to 18 degrees C. Furthermore, incubation at 4 degrees C totally prevented this DNA movement, even if a large part of the DNA had already exited the capsid.
D000327:Adsorption; D001425:Bacterial Outer Membrane Proteins; D004279:DNA, Viral / Q000378:metabolism*; D004926:Escherichia coli / Q000032:analysis; D008081:Liposomes; D008274:Magnesium / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism*; D011700:Putrescine / Q000494:pharmacology; D011991:Receptors, Virus / Q000378:metabolism*; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D013604:T-Phages / Q000378:metabolism*; D013696:Temperature
D001425:Bacterial Outer Membrane Proteins; D004279:DNA, Viral; D008081:Liposomes; D008565:Membrane Proteins; D011991:Receptors, Virus; D013096:Spermine; D008274:Magnesium; D013095:Spermidine; D011700:Putrescine