title: The msbB mutant of Neisseria meningitidis strain NMB has a defect in lipooligosaccharide assembly and transport to the outer membrane.
abstract: A deletion-insertion mutation in msbB, a gene that encodes a lipid A acyltransferase, was introduced into encapsulated Neisseria meningitidis serogroup B strain NMB and an acapsular mutant of the same strain. These mutants were designated NMBA11K3 and NMBA11K3cap-, respectively. Neither lipooligosaccharide (LOS) nor lipid A could be isolated from NMBA11K3 although a number of techniques were tried, but both were easily extracted from NMBA11K3cap-. Immunoelectron microscopy using monoclonal antibody (MAb) 6B4, which recognizes the terminal Galbeta1-4GlcNAc of LOS, demonstrated that NMB, NMBcap-, and NMBA11K3cap- expressed LOS circumferentially, while MAb 6B4 did not bind to the surface of NMBA11K3. However, cytoplasmic staining of NMBA11K3 with MAb 6B4 was a consistent observation. Mass-spectrometric analyses demonstrated that the relative amounts of the lipid A-specific C12:0 3-OH and C14:0 3-OH present in the membrane preparations (MP) from NMBA11K3 were substantially decreased (25- and 23-fold, respectively) compared to the amount in MP from its parent strain, NMB. Western blot analyses of MP from NMBA11K3 demonstrated that the levels of porin in the outer membrane of NMBA11K3 were also substantially decreased. These studies suggest that the lipid A acylation defect in encapsulated NMBA11K3 influences the assembly of the lipid A and consequently the incorporation of porin in the outer membrane.
authors.affiliation: Department of Microbiology, University of Iowa, Iowa City 52242, USA.
authors.forename: Deborah M B
authors.initials: DM
authors.lastname: Post
authors.forename: Margaret R
authors.initials: MR
authors.lastname: Ketterer
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Phillips
authors.forename: Bradford W
authors.initials: BW
authors.lastname: Gibson
authors.forename: Michael A
authors.initials: MA
authors.lastname: Apicella
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D005227:Fatty Acids / Q000032:analysis; D017353:Gene Deletion; D008050:Lipid A / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D013058:Mass Spectrometry; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D016253:Microscopy, Immunoelectron; D009154:Mutation; D009345:Neisseria meningitidis / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D016133:Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D005227:Fatty Acids; D008050:Lipid A; D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; D000217:Acyltransferases