In silico analysis of usher encoding genes in Klebsiella pneumoniae and characterization of their role in adhesion and colonization

PloS One
Fida KhaterChristiane Forestier

Abstract

Chaperone/usher (CU) assembly pathway is used by a wide range of Enterobacteriaceae to assemble adhesive surface structures called pili or fimbriae that play a role in bacteria-host cell interactions. In silico analysis revealed that the genome of Klebsiella pneumoniae LM21 harbors eight chromosomal CU loci belonging to γκп and ϭ clusters. Of these, only two correspond to previously described operons, namely type 1 and type 3-encoding operons. Isogenic usher deletion mutants of K. pneumoniae LM21 were constructed for each locus and their role in adhesion to animal (Intestine 407) and plant (Arabidopsis thaliana) cells, biofilm formation and murine intestinal colonization was investigated. Type 3 pili usher deleted mutant was impaired in all assays, whereas type 1 pili usher deleted mutant only showed attenuation in adhesion to plant cells and in intestinal colonization. The LM21ΔkpjC mutant was impaired in its capacity to adhere to Arabidopsis cells and to colonize the murine intestine, either alone or in co-inoculation experiments. Deletion of LM21kpgC induced a significant decrease in biofilm formation, in adhesion to animal cells and in colonization of the mice intestine. The LM21∆kpaC and LM21∆kpeC mutants were only attenua...Continue Reading

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Datasets Mentioned

BETA
PRJEB7075
ABR79894

Methods Mentioned

BETA
PCRs
PCR
Protein Assay
gene knock-out

Software Mentioned

assembler Genomics Workbench
AlienTrimmer
Entrez
Microscope
MAFFT
trimal
Magnifying Genomes ( MaGe )
PhyML
BLASTp
Mauve Contig Mover

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