title: Effect of Bdellovibrio bacteriovorus infection on the phosphoenolpyruvate:sugar phosphotransferase system in Escherichia coli: evidence for activation of cytoplasmic proteolysis.
abstract: Intact cells of Bdellovibrio bacteriovorus strain 109J were found to be incapable of taking up 14C-methyl alpha-glucoside, mannitol or fructose, and extracts derived from these cells exhibited negligible activities of the protein components of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Escherichia coli strain ML35 cells exhibited high in vivo sugar uptake activities that were progressively lost over a period of 2 h at 30 degrees C following the entry of B. bacteriovorus into the periplasm of E. coli. In vitro complementation assays revealed that the E. coli PTS enzymes, enzyme I, HPr, and the glucose- and mannitol-specific enzymes II, were all lost almost in parallel with the disappearance of uptake activity. Thus, loss of activity in vivo was not due to membrane leakiness, energy depletion, or preferential inhibition or inactivation of any one protein component of the PTS. Instead, loss of PTS activity was attributed to digestion of the protein constituents of the system by proteases present in the cytoplasm of the host cell after bdellovibrio entry. Both ethylenediaminetetraacetate and phenylmethylsulphonyl fluoride partially protected against inactivation in vitro, and the two inhibitors together gave full protection, suggesting that both metallo- and seryl-proteases were responsible for the inactivation. Protease activity increased progressively with time following bdellovibrio entry and appeared to degrade the E. coli PTS enzymes in vivo. Preliminary evidence suggested that the proteases responsible for PTS enzyme degradation may be encoded by the B. bacteriovorus chromosome.
authors.affiliation: Department of Biology, University of California, San Diego, La Jolla 92093-0116.
authors.forename: A J
authors.initials: AJ
authors.lastname: Romo
authors.forename: E G
authors.initials: EG
authors.lastname: Ruby
authors.forename: M H
authors.initials: MH
authors.lastname: Saier
mesh_terms: D001501:Bdellovibrio / Q000378:metabolism / Q000472:pathogenicity*; D003864:Depression, Chemical; D004926:Escherichia coli / Q000378:metabolism*; D066298:In Vitro Techniques; D008353:Mannitol / Q000378:metabolism; D008757:Methylglucosides / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D010728:Phosphoenolpyruvate / Q000378:metabolism; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000378:metabolism*; D010770:Phosphotransferases
chemical_list: D008757:Methylglucosides; D008353:Mannitol; D010728:Phosphoenolpyruvate; D010770:Phosphotransferases; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D010447:Peptide Hydrolases