title: GNA33 of Neisseria meningitidis is a lipoprotein required for cell separation, membrane architecture, and virulence.
abstract: GNA33 is a membrane-bound lipoprotein with murein hydrolase activity that is present in all Neisseria species and well conserved in different meningococcal isolates. The protein shows 33% identity to a lytic transglycolase (MltA) from Escherichia coli and has been shown to be involved in the degradation of both insoluble murein sacculi and unsubstituted glycan strands. To study the function of the gene and its role in pathogenesis and virulence, a knockout mutant of a Neisseria meningitidis serogroup B strain was generated. The mutant exhibited retarded growth in vitro. Transmission electron microscopy revealed that the mutant grows in clusters which are connected by a continuous outer membrane, suggesting a failure in the separation of daughter cells. Moreover, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of culture supernatant revealed that the mutant releases several proteins in the medium. The five most abundant proteins, identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis, belong to the outer membrane protein family. Finally, the mutant showed an attenuated phenotype, since it was not able to cause bacteremia in the infant rat model. We conclude that GNA33 is a highly conserved lipoprotein which plays an important role in peptidoglycan metabolism, cell separation, membrane architecture, and virulence.
authors.affiliation: IRIS, Chiron Vaccines, 53100 Siena, Italy.
authors.forename: Jeannette
authors.initials: J
authors.lastname: Adu-Bobie
authors.forename: Pietro
authors.initials: P
authors.lastname: Lupetti
authors.forename: Brunella
authors.initials: B
authors.lastname: Brunelli
authors.forename: Dan
authors.initials: D
authors.lastname: Granoff
authors.forename: Nathalie
authors.initials: N
authors.lastname: Norais
authors.forename: Germano
authors.initials: G
authors.lastname: Ferrari
authors.forename: Guido
authors.initials: G
authors.lastname: Grandi
authors.forename: Rino
authors.initials: R
authors.lastname: Rappuoli
authors.forename: Mariagrazia
authors.initials: M
authors.lastname: Pizza
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D021461:Animals, Outbred Strains; D016470:Bacteremia / Q000382:microbiology / Q000503:physiopathology; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D017353:Gene Deletion; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008589:Meningococcal Infections / Q000382:microbiology / Q000503:physiopathology; D008854:Microscopy, Electron; D038541:Neisseria meningitidis, Serogroup B / Q000737:chemistry / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D008074:Lipoproteins; D016695:Glycosyltransferases; C085441:MltA protein, bacteria