title: Ght protein of Neisseria meningitidis is involved in the regulation of lipopolysaccharide biosynthesis.
abstract: Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria and is responsible for the barrier function of this membrane. A ght mutant of Neisseria meningitidis that showed increased sensitivity to hydrophobic toxic compounds, suggesting a breach in this permeability barrier, was previously described. Here, we assessed whether this phenotype was possibly caused by a defect in LPS transport or synthesis. The total amount of LPS appeared to be drastically reduced in a ght mutant, but the residual LPS was still detected at the cell surface, suggesting that LPS transport was not impaired. The ght mutant was rapidly overgrown by pseudorevertants that produced normal levels of LPS. Genetic analysis of these pseudorevertants revealed that the lpxC gene, which encodes a key enzyme in LPS synthesis, was fused to the promoter of the upstream-located pilE gene, resulting in severe lpxC overexpression. Analysis of phoA and lacZ gene fusions indicated that Ght is an inner membrane protein with an N-terminal membrane anchor and its bulk located in the cytoplasm, where it could potentially interact with LpxC. Cell fractionation experiments indeed indicated that Ght tethers LpxC to the membrane. We suggest that Ght regulates LPS biosynthesis by affecting the activity of LpxC. Possibly, this mechanism acts in the previously observed feedback inhibition of LPS synthesis that occurs when LPS transport is hampered.
authors.affiliation: Department of Molecular Microbiology and Institute of Biomembranes, Utrecht University, Utrecht, Netherlands.
authors.forename: Florian
authors.initials: F
authors.lastname: Putker
authors.forename: Andreas
authors.initials: A
authors.lastname: Grutsch
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
authors.forename: Martine P
authors.initials: MP
authors.lastname: Bos
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D008070:Lipopolysaccharides / Q000096:biosynthesis*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D011485:Protein Binding; D013489:Suppression, Genetic
chemical_list: D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D008565:Membrane Proteins