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Expression data from Burkholderia cepacia IST408 bceE and bceF mutants

ABSTRACT: The bacterial tyrosine (BY) kinase family comprises the major group of bacterial enzymes endowed with tyrosine kinase activity. We have previously shown that the BceF protein from Burkholderia cepacia IST408 belongs to this BY kinase family and is involved in the biosynthesis of the exopolysaccharide cepacian. However, little is known about the extent of regulation in the cell by this kinase protein. In order to examine this, we performed a comparative transcriptome profile between the bceF mutant and the wild-type B. cepacia IST408. As a control, the transcriptome of another exopolysaccharide-deficient mutant, which had a mutation in the bceE gene, was also determined.

ORGANISM(S): Burkholderia cepacia Burkholderia cenocepacia J2315 Burkholderia multivorans ATCC 17616

PROVIDER: GSE38183 | GEO | 2012/12/01

SECONDARY ACCESSION(S): PRJNA167335

REPOSITORIES: GEO

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Expression data from Burkholderia cepacia IST408 bceE and bceF mutants

Project description:The bacterial tyrosine (BY) kinase family comprises the major group of bacterial enzymes endowed with tyrosine kinase activity. We have previously shown that the BceF protein from Burkholderia cepacia IST408 belongs to this BY kinase family and is involved in the biosynthesis of the exopolysaccharide cepacian. However, little is known about the extent of regulation in the cell by this kinase protein. In order to examine this, we performed a comparative transcriptome profile between the bceF mutant and the wild-type B. cepacia IST408. As a control, the transcriptome of another exopolysaccharide-deficient mutant, which had a mutation in the bceE gene, was also determined. For expression profiling, overnight cultures of the Burkholderia cepacia IST408 wild-type strain and the isogenic mutants bceE::Tp and bceF::Tp grown in LB medium were diluted to an initial OD640 nm of 0.1 into S medium. Triplicate samples were cultured at 30ÂºC with 250 r.p.m. agitation for 12 h, and RNA was extracted from the bacterial isolates. DNA-based probe-selection was performed using a CEL file derived from a hybridization of genomic DNA, and Xspecies DNA hyb CDF batchmaker v3.2, available here: http://affymetrix.arabidopsis.info/xspecies/. An optimal signal cutoff of 80 and minimum number of probes per probe set of 7 were determined empirically, resulting in the masking of 24223 of the original 226576 probes on the array and the exclusion of 156 probe sets. The new cdf file created (Bcc1sa520656F_Xspecies.CEL80.cdf) was used in DNA-Chip Analyzer 2008 for the subsequent gene expression analyses.

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Overexpression of a histone-like nucleoid structuring protein in Burkholderia multivorans affects several traits dependent on the cell envelope

Project description:Burkholderia cepacia complex bacteria comprise opportunistic pathogens causing chronic respiratory infections in cystic fibrosis (CF) patients. These microorganisms produce exopolysaccharides which are considered virulence determinants. Here, we report the isolation of an evolved nonmucoid variant (17616nmv) after the ancestor, Burkholderia multivorans ATCC 17616, was subjected to prolonged stationary phase. Lack of exopolysaccharide cepacian biosynthesis in the variant was correlated with downregulation of bce genes expression. Furthermore, genome sequence of the variant identified the transposition of a mobile element of the IS406 family into the promoter region of an hns-like gene (Bmul_0158) encoding a histone-like nucleoid structuring protein, a known global transcriptional repressor. This IS element upregulated the expression of Bmul_0158 gene. Transcriptome analysis identified the global effects of this mutation on gene expression, with major changes in motility, pili synthesis, type VI secretion, and chromosome associated functions. Concomitant with these differences, the nonmucoid variant displays reduced adherence to a CF lung bronchial cell line, reduced surface hydrophobicity, forms smaller cellular aggregates, but has an increase in swimming and swarming motilities. Finally, bioinformatic analysis led to the identification of various genomic regions, possibly acquired by horizontal gene transfer, which were transcriptionally repressed by the increased expression of hns gene in the nonmucoid variant. Taken together, our results revealed a significant role for this H-NS protein in the regulation of B. multivorans persistence- and virulence-associated genes.

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