title: Molecular analysis of the Acinetobacter baumannii biofilm-associated protein.
abstract: Acinetobacter baumannii is a multidrug-resistant pathogen associated with hospital outbreaks of infection across the globe, particularly in the intensive care unit. The ability of A. baumannii to survive in the hospital environment for long periods is linked to antibiotic resistance and its capacity to form biofilms. Here we studied the prevalence, expression, and function of the A. baumannii biofilm-associated protein (Bap) in 24 carbapenem-resistant A. baumannii ST92 strains isolated from a single institution over a 10-year period. The bap gene was highly prevalent, with 22/24 strains being positive for bap by PCR. Partial sequencing of bap was performed on the index case strain MS1968 and revealed it to be a large and highly repetitive gene approximately 16 kb in size. Phylogenetic analysis employing a 1,948-amino-acid region corresponding to the C terminus of Bap showed that BapMS1968 clusters with Bap sequences from clonal complex 2 (CC2) strains ACICU, TCDC-AB0715, and 1656-2 and is distinct from Bap in CC1 strains. By using overlapping PCR, the bapMS1968 gene was cloned, and its expression in a recombinant Escherichia coli strain resulted in increased biofilm formation. A Bap-specific antibody was generated, and Western blot analysis showed that the majority of A. baumannii strains expressed an ∼200-kDa Bap protein. Further analysis of three Bap-positive A. baumannii strains demonstrated that Bap is expressed at the cell surface and is associated with biofilm formation. Finally, biofilm formation by these Bap-positive strains could be inhibited by affinity-purified Bap antibodies, demonstrating the direct contribution of Bap to biofilm growth by A. baumannii clinical isolates.
authors: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Queensland, Australia.
authors: H M Sharon
authors: HM
authors: Goh
authors: Scott A
authors: SA
authors: Beatson
authors: Makrina
authors: M
authors: Totsika
authors: Danilo G
authors: DG
authors: Moriel
authors: Minh-Duy
authors: MD
authors: Phan
authors: Jan
authors: J
authors: Szubert
authors: Naomi
authors: N
authors: Runnegar
authors: Hanna E
authors: HE
authors: Sidjabat
authors: David L
authors: DL
authors: Paterson
authors: Graeme R
authors: GR
authors: Nimmo
authors: Jeffrey
authors: J
authors: Lipman
authors: Mark A
authors: MA
authors: Schembri
mesh_terms: D000151:Acinetobacter Infections / Q000453:epidemiology* / Q000382:microbiology; D040981:Acinetobacter baumannii / Q000235:genetics*; D000704:Analysis of Variance; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D018441:Biofilms; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D016000:Cluster Analysis; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011793:Queensland / Q000453:epidemiology; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D008565:Membrane Proteins
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