F exclusion of bacteriophage T7 occurs at the cell membrane.
The F plasmid PifA protein, known to be the cause of F exclusion of bacteriophage T7, is shown to be a membrane-associated protein. No transmembrane domains of PifA were located. In contrast, T7 gp1.2 and gp10, the two phage proteins that trigger phage exclusion, are both soluble cytoplasmic proteins. The Escherichia coli FxsA protein, which, at higher concentrations than found in wild-type cells, protects T7 from exclusion, is shown to interact with PifA. FxsA is a polytopic membrane protein with four transmembrane segments and a long cytoplasmic C-terminal tail. This tail is not important in alleviating F exclusion and can be deleted; in contrast, the fourth transmembrane segment of FxsA is critical in allowing wild-type T7 to grow in the presence of F PifA. These data suggest that the primary event that triggers the exclusion process occurs at the cytoplasmic membrane and that FxsA sequesters PifA so that membrane damage is minimized.
Molecular Genetics and Microbiology,University of Texas, Austin 78712, USA.
Xiaogang
X
Cheng
WenFang
W
Wang
Ian J
IJ
Molineux
D017123:Bacteriophage T7 / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000821:virology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005144:F Factor / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D012097:Repressor Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D012097:Repressor Proteins; D014764:Viral Proteins; C121882:fxsA protein, E coli; C120334:gene 2 product, bacteriophage T7