title: The Omp85 protein of Neisseria meningitidis is required for lipid export to the outer membrane.
abstract: In Gram-negative bacteria, lipopolysaccharide and phospholipid biosynthesis takes place at the inner membrane. How the completed lipid molecules are subsequently transported to the outer membrane remains unknown. Omp85 of Neisseria meningitidis is representative for a family of outer membrane proteins conserved among Gram-negative bacteria. We first demonstrated that the omp85 gene is co-transcribed with genes involved in lipid biosynthesis, suggesting an involvement in lipid assembly. A meningococcal strain was constructed in which Omp85 expression could be switched on or off through a tac promoter-controlled omp85 gene. We demonstrated that the presence of Omp85 is essential for viability. Depletion of Omp85 leads to accumulation of electron-dense amorphous material and vesicular structures in the periplasm. We demonstrated, by fractionation of inner and outer membranes, that lipopolysaccharide and phospholipids mostly disappeared from the outer membrane and instead accumulated in the inner membrane, upon depletion of Omp85. Omp85 depletion did not affect localization of integral outer membrane proteins PorA and Opa. These results provide compelling evidence for a role for Omp85 in lipid transport to the outer membrane.
authors.affiliation: Research Unit in Molecular Biology (URBM), University of Namur (FUNDP), Rue de Bruxelles, 61, 5000 Namur, Belgium. stephanie.genevrois@fundp.ac.be
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Genevrois
authors.forename: Liana
authors.initials: L
authors.lastname: Steeghs
authors.forename: Paul
authors.initials: P
authors.lastname: Roholl
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Letesson
authors.forename: Peter
authors.initials: P
authors.lastname: van der Ley
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D050356:Lipid Metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000378:metabolism* / Q000648:ultrastructure; D010743:Phospholipids / Q000378:metabolism; D017433:Protein Structure, Secondary; D014158:Transcription, Genetic
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; C115176:Omp85 protein, Neisseria; D010743:Phospholipids