Genetic analysis of MraY inhibition by the phiX174 protein E.
Protein E, the lysis protein of bacteriophage phiX174, is a specific inhibitor of MraY, the phospho-MurNAc-pentapeptide translocase that catalyzes the synthesis of lipid I in the conserved pathway for peptidoglycan biosynthesis. The original evidence for this inhibition was the isolation of two spontaneous E-resistance mraY mutants. Here we report further genetic studies aimed at dissecting the interaction between E and MraY, using a genetic strategy that is facile, rapid, and does not depend on the availability of purified E, purified MraY, or its substrates. This system relies on the ability of mraY or its enzymatically inactive D267N allele to protect cells from lysis after induction of a chimeric lambda :: E prophage. Using this approach, the MraY protein from Bacillus subtilis, which shares 43% sequence identity with the Escherichia coli enzyme, was found to interact weakly, if at all, with E. A potential E binding site defined by transmembrane domains 5 and 9 has been identified by isolating more mraY mutants resistant to E inhibition. Genetic analysis indicates that these E-resistant alleles fall into three classes on the basis of the affinity of the encoded proteins for MraY.
Zheng
Yi
Y
Department of Biochemistry and Biophysics, Texas A & M University, College Station, Texas 77843-2128, USA.
Struck
Douglas K
DK
Bernhardt
Thomas G
TG
Young
Ry
R
D001412:Bacillus subtilis / Q000502:physiology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D001433:Bacteriolysis; D002478:Cells, Cultured; D004926:Escherichia coli / Q000502:physiology*; D010457:Peptidoglycan / Q000378:metabolism; D010957:Plasmids; D014166:Transferases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups); D014170:Transformation, Genetic; D014764:Viral Proteins / Q000235:genetics*
D001426:Bacterial Proteins; C029555:E protein, bacteriophage X174; D010457:Peptidoglycan; D014764:Viral Proteins; D014166:Transferases; D017855:Transferases (Other Substituted Phosphate Groups); C079803:mraY protein, Bacteria
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