Identification of Klebsiella pneumoniae genes involved in intestinal colonization and adhesion using signature-tagged mutagenesis.
Klebsiella pneumoniae is an opportunistic pathogen responsible for nosocomial infections that initially colonize the intestinal tract of patients. Signature-tagged mutagenesis was used to identify genes required for this function. A library of 2,200 mutants was analyzed for the inability of the mutants to survive in a murine model of intestinal colonization and to adhere to human intestinal cells (Int-407) in vitro. Twenty-nine attenuated mutants were selected for further analyses after competition assays against the wild-type strain. Whatever the screening model, most of the transposon insertions occurred in genes involved in metabolic pathways, membrane transport, DNA metabolism, transcriptional regulation, and unknown functions. Only one mutant was attenuated in both the murine colonization and the in vitro adhesion models, and the sequence disrupted by the transposon had homology to adhesin-encoding genes of Haemophilus sp.
Laboratoire de Bactériologie, Université d'Auvergne, Faculté de Pharmacie, 63000 Clermont-Ferrand, France.
Nathalie
N
Maroncle
Damien
D
Balestrino
Chantal
C
Rich
Christiane
C
Forestier
D018829:Adhesins, Bacterial / Q000235:genetics; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics; D004064:Digestive System / Q000382:microbiology*; D004195:Disease Models, Animal; D029968:Escherichia coli Proteins / Q000235:genetics; D005260:Female; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial / Q000502:physiology*; D024981:Glycogen Phosphorylase / Q000235:genetics; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007710:Klebsiella Infections / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000254:growth & development; D051379:Mice; D016297:Mutagenesis, Site-Directed; D051922:PII Nitrogen Regulatory Proteins; D018272:Porins / Q000235:genetics; D011494:Protein Kinases; D027981:Symporters / Q000235:genetics; D015534:Trans-Activators; D014157:Transcription Factors
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C088688:FucP protein, E coli; D047908:Intracellular Signaling Peptides and Proteins; D051922:PII Nitrogen Regulatory Proteins; D018272:Porins; D027981:Symporters; D015534:Trans-Activators; D014157:Transcription Factors; C110884:fmdD protein, Methylophilus methylotrophus; C028640:fructose-induced HPr-like protein, bacteria; D024981:Glycogen Phosphorylase; D011494:Protein Kinases; C488924:fruB protein, E coli; C103622:FucA protein, bacteria; C491231:fucA protein, E coli; D005634:Fructose-Bisphosphate Aldolase