title: Reduced DNA binding and uptake in the absence of DsbA1 and DsbA2 of Neisseria meningitidis due to inefficient folding of the outer-membrane secretin PilQ.
abstract: DsbA ensures the correct folding of many exported bacterial proteins by forming intramolecular disulphide bonds in the bacterial periplasm. The pathogen Neisseria meningitidis is unusual in its possession of three different dsbA genes (dsbA1, dsbA2 and dsbA3), encoding two membrane-anchored (DsbA1 and DsbA2) and one periplasmic (DsbA3) thiol-disulphide oxidoreductase enzymes. In this study, the involvement of DsbA1 and DsbA2 in natural competence was confirmed and attributed to events in the early stages of the transformation process. Strains lacking both DsbA1 and DsbA2 were reduced in competence as a result of decreased DNA binding and uptake. Overexpression of DsbA3 could not overcome this defect, suggesting differences in substrate specificity and protein-folding abilities between the DsbA homologues. Competence in Neisseria is dependent on the expression of type IV pili, which are extruded and retracted through the outer-membrane secretin PilQ. Both DsbA1 and DsbA2 were able to specifically bind PilQ in solid-phase overlay assays. Consistent with this, deletion of both dsbA1 and dsbA2 resulted in reduced levels of PilQ, confirming inefficient folding of PilQ, while pilus expression was apparently unaffected. The secretin PilQ is involved in DNA binding and transport as well as pilus biogenesis, and the defect in PilQ folding resulting from the absence of DsbA1 and DsbA2 is revealed in the observed decreased DNA binding and uptake.
authors.affiliation: Molecular Infectious Diseases Group, Department of Paediatrics, Imperial College, St Mary's Campus, Norfolk Place, London W2 1PG, UK.
authors.forename: Sunita
authors.initials: S
authors.lastname: Sinha
authors.affiliation: Centre for Molecular Biology and Neuroscience and Institute of Microbiology, University of Oslo, Norway.
authors.forename: Ole Herman
authors.initials: OH
authors.lastname: Ambur
authors.affiliation: Molecular Infectious Diseases Group, Department of Paediatrics, Imperial College, St Mary's Campus, Norfolk Place, London W2 1PG, UK.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Langford
authors.affiliation: Centre for Molecular Biology and Neuroscience and Institute of Microbiology, Rikshospitalet-Radiumhospitalet Medical Centre, Oslo, Norway.|Centre for Molecular Biology and Neuroscience and Institute of Microbiology, University of Oslo, Norway.
authors.forename: Tone
authors.initials: T
authors.lastname: Tønjum
authors.affiliation: Molecular Infectious Diseases Group, Department of Paediatrics, Imperial College, St Mary's Campus, Norfolk Place, London W2 1PG, UK.
authors.forename: J Simon
authors.initials: JS
authors.lastname: Kroll
mesh_terms: D004269:DNA, Bacterial / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000378:metabolism / Q000502:physiology; D017353:Gene Deletion; D018628:Gene Dosage; D016254:Mutagenesis, Insertional; D009345:Neisseria meningitidis / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*; D014169:Transformation, Bacterial / Q000235:genetics / Q000502:physiology*
chemical_list: D004269:DNA, Bacterial; C084295:pilQ protein, bacteria; D036781:Fimbriae Proteins; D019704:Protein Disulfide-Isomerases