PMID:10760296

Citation	Bernhardt, TG, Roof, WD and Young, R (2000) Genetic evidence that the bacteriophage phi X174 lysis protein inhibits cell wall synthesis. Proc. Natl. Acad. Sci. U.S.A. 97:4297-302
Abstract	Protein E, a 91-residue membrane protein of phiX174, causes lysis of the host in a growth-dependent manner reminiscent of cell wall antibiotics, suggesting E acts by inhibiting peptidoglycan synthesis. In a search for the cellular target of E, we previously have isolated recessive mutations in the host gene slyD (sensitivity to lysis) that block the lytic effects of E. The role of slyD, which encodes a FK506 binding protein-type peptidyl-prolyl cis-trans isomerase, is not fully understood. However, E mutants referred to as Epos (plates on slyD) lack a slyD requirement, indicating that slyD is not crucial for lysis. To identify the gene encoding the cellular target, we selected for survivors of Epos. In this study, we describe the isolation of dominant mutations in the essential host gene mraY that result in a general lysis-defective phenotype. mraY encodes translocase I, which catalyzes the formation of the first lipid-linked intermediate in cell wall biosynthesis. The isolation of these lysis-defective mutants supports a model in which translocase I is the cellular target of E and that inhibition of cell wall synthesis is the mechanism of lysis.
Links	PubMed PMC18234
Keywords	Alleles; Bacteriophage phi X 174/genetics; Bacteriophage phi X 174/metabolism; Carrier Proteins/genetics; Cell Wall/metabolism; Chromosome Mapping; Chromosomes, Bacterial; Escherichia coli/genetics; Escherichia coli/virology; Escherichia coli Proteins; Genes, Essential; Mutation; Peptidylprolyl Isomerase/genetics; Phenotype; Polymerase Chain Reaction; Viral Proteins/genetics; Viral Proteins/physiology
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BPPHS:E		GO:0044659: cytolysis by virus of host cell	ECO:0000314:		P		Figure 2A: phage E protein lyses E. coli cells expressing wildtype slyD and mraY - absorbance decreases over time	complete CACAO 10541
BPPHS:E		GO:0039635: suppression by virus of host peptidoglycan biosynthetic process	ECO:0000316:	UniProtKB:P0A6W3	P		Figure 2D: more time was required for phage E protein to lyse cells in E. coli expressing additional mraY (produced by plasmids, represented by square) than in E. coli expressing normal amounts of mraY (represented by circle) - shows that E targets mraY - mraY is known to catalyze a reaction involved in peptidoglycan synthesis (reference 39)	complete CACAO 10546
BPPHS:E	involved_in	GO:0044659: cytolysis by virus of host cell	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
BPPHS:E	involved_in	GO:0039635: suppression by virus of host peptidoglycan biosynthetic process	ECO:0000316: genetic interaction evidence used in manual assertion	UniProtKB:P0A6W3	P		Seeded From UniProt	complete
BPPHS:E		GO:0039640: cytolysis by virus via suppression of host peptidoglycan biosynthetic process	ECO:0000316:	UniProtKB:P0A6W3	P		Fig 2D – E-mediated lysis occurs in wt host cells (circle), is delayed in cells overexpressing MraY (MraY expressed from medium copy plasmid, square), and doesn’t occur in cells overexpressing an MraY mutant (MraY mutant expressed from same medium copy plasmid, triangle). Suggests that E causes host lysis by inhibiting MraY. MraY is an enzyme involved in PG biosynthesis.	complete CACAO 11831

References

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