Protein-mediated DNA transfer into liposomes.
The transfer of a foreign genome into a bacterium by means of phage infection is a very efficient but poorly understood process. To analyse the mechanism of phage DNA transfer at a molecular level, we have reconstituted FhuA, the receptor for phage T5 in the outer membrane of Escherichia coli, into unilamellar vesicles made of natural phospholipids. Cryoelectron microscopy studies showed that the binding of the phage to FhuA triggered the transfer of its double-stranded DNA (121000 bp) into the proteoliposomes. DNA was entrapped within vesicles with a diameter ranging from 70 to 150 nm. The DNA appeared to be densely packed, but its presence did not alter the morphology of the liposomes, suggesting no DNA-lipid interactions. These liposomes represent an attractive model system for studying the mechanisms of DNA transport and condensation. They may also serve as alternative vehicles for the transfer of foreign genes into eukaryotic cells.
Lambert
O
O
Institut Curie, Section de Recherche, UMR 168 CNRS et LRC-CEA 8, Paris, France.
Plançon
L
L
Rigaud
J L
JL
Letellier
L
L
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D029968:Escherichia coli Proteins; D018014:Gene Transfer Techniques; D008081:Liposomes; D011991:Receptors, Virus / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C050900:FhuA protein, E coli; D008081:Liposomes; D011991:Receptors, Virus