A membrane protein is required for bacteriophage c2 infection of Lactococcus lactis subsp. lactis C2.
Phage-resistant mutants, isolated from cultures of Lactococcus lactis subsp. lactis C2 infected with phage c2, did not form plaques but bound phage normally. The mutants were sensitive to another phage, sk1, although the number of plaques was reduced approximately 56% and the plaques were four times smaller. Binding to phage sk1 was reduced about 10%. Another group of phage-resistant mutants, isolated from cultures infected with phage sk1, bound normally to both phages c2 and sk1 but did not form plaques with either phage. Carbohydrate analyses by gas chromatography of the cell walls showed no significant differences in saccharide compositions between the wild-type and phage-resistant cells. However, a difference was observed in the interactions of the phage with the cytoplasmic membranes. Membranes from the wild-type cells, but not mutant cells, inactivated phage c2. Phage sk1 was not inactivated by membrane from either strain. Treatment of wild-type membranes with proteinase K eliminated the ability of the membrane to inactivate the phage, whereas treatment with mutanolysin had no effect. On the basis of this ability to inactivate the phage, a membrane protein was partially purified by gel filtration and ion-exchange chromatography. Under nondenaturing conditions, the phage-inactivating protein has an apparent Mr of approximately 350,000. The protein has an apparent subunit size of 32 kDa, which suggests that it normally exists as a multimer with 10 to 12 subunits or in association with other membrane components. It is proposed that this protein is required for phage c2 infection.
Valyasevi
R
R
Department of Microbiology, Oregon State University, Corvallis 97331-3804.
Sandine
W E
WE
Geller
B L
BL
D001426:Bacterial Proteins / Q000302:isolation & purification*; D001435:Bacteriophages / Q000235:genetics / Q000254:growth & development* / Q000502:physiology; D002462:Cell Membrane / Q000382:microbiology; D013294:Lactococcus lactis / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D008565:Membrane Proteins / Q000302:isolation & purification / Q000502:physiology*; D008970:Molecular Weight; D010948:Viral Plaque Assay; D014775:Virus Activation
D001426:Bacterial Proteins; D008565:Membrane Proteins