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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Expression Data from Acinetobacter baumannii strain ATCC 17978 and isogenic lpsB mutant cells

ABSTRACT: lpsB encodes a glycosyltransferase involved in lipopolysaccharide (LPS) synthesis. LPS is a major component of the Gram-negative bacterial outer membranes. We used custom-made Affymetrix A. baumannii strain ATCC 17978 derived GeneChips to compare the gene expression properties of wild type and isogenic lpsB mutant cells. Two mutants were evaluated; A. baumannii strain 5A7 is a ATCC 17978 derivative harboring a transposon (Tn5) within lpsB (A1S_0430 locus); A. baumannii strain containing a deletion of lpsB (A1S_0430). A. baumannii strain ATCC 17978, 5A7 (lpsB:Tn5) or IH1∆lpsB (∆lpsB) were grown to mid-exponential phase growth, total bacterial RNA was isolated and subjected to GeneChip hybridization and analysis. We sought to determine the transcription profile of lpsB mutated cells.

ORGANISM(S): Acinetobacter baumannii

SUBMITTER: Paul Dunman

PROVIDER: E-GEOD-40771 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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An Ribonuclease T2 Family Protein Modulates Acinetobacter baumannii Abiotic Surface Colonization

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Expression Data from Acinetobacter baumannii strain ATCC 17978 and isogenic lpsB mutant cells

Project description:lpsB encodes a glycosyltransferase involved in lipopolysaccharide (LPS) synthesis. LPS is a major component of the Gram-negative bacterial outer membranes. We used custom-made Affymetrix A. baumannii strain ATCC 17978 derived GeneChips to compare the gene expression properties of wild type and isogenic lpsB mutant cells. Two mutants were evaluated; A. baumannii strain 5A7 is a ATCC 17978 derivative harboring a transposon (Tn5) within lpsB (A1S_0430 locus); A. baumannii strain containing a deletion of lpsB (A1S_0430).

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Project description:In this study the transcriptomic changes occurring in A. baumannii ATCC 17978 in response to shock treament using the biocide chlorhexdine were determined. A. baumannii ATCC 17978 cells were grown at 37Â°C with shaking (200rpm) in 35mL cultures in MH broth to OD600=0.75, at which time they were split into 15mL cultures. One 15mL sample was treated with 4Âµg/mL chlorhexidine (0.5x MIC), whereas the other was not treated and used as a reference. Cultures were allowed to grow for a further 30 mins (to an average final OD600= ~1.35 for untreated cells and 1.25 for chlorhexidine treated samples), when cells were harvested by centrifugation and immediately suspended in Trizol reagent (Invitrogen). Total RNA was extracted using the PureLinkTM Micro-to-Midi Total RNA Purification kit (Invitrogen), incorporating an on-column DNAseI (Invitrogen) digestion. The experiment included using four grids of a custom Agilent 8 x 15000 spot microarray. Three biological replicates and one flip dye experiment.

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Comparative transcriptional analysis of Acinetobacter baumannii strain ATCC 17978 and strain 17978hm

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Comparative transcriptional analysis of Acinetobacter baumannii strain ATCC 17978 and strain 17978hm

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Colistin-resistant, lipopolysaccharide-deficient Acinetobacter baumannii responds to lipopolysaccharide loss through increased expression of genes involved in the synthesis and transport of lipoproteins, phospholipids and poly-beta-1,6-N-acetylglucosamine

Project description:We recently demonstrated that colistin resistance in Acinetobacter baumannii can result from mutational inactivation of genes essential for lipid A biosynthesis. Consequently, strains harboring these mutations are unable to produce the major Gram negative bacterial surface component, lipopolysaccharide (LPS). To understand how A. baumannii compensates for the lack of LPS, we compared the transcriptional profile of the A. baumannii type strain ATCC19606, to that of an isogenic, LPS-deficient, lpxA mutant strain. Analysis of the expression profiles indicated that the LPS-deficient strain showed increased expression of many genes involved in cell envelope and membrane biogenesis. In particular, up-regulated genes included those involved in the Lol lipoprotein transport system and the Mla-retrograde phospholipid transport system. In addition, genes involved in the synthesis and transport of poly-beta-1,6-N-acetylglucosamine (PNAG) were also up-regulated and a corresponding increase in PNAG production was observed. The LPS-deficient strain also exhibited reduced expression of genes predicted to encode the fimbrial subunit FimA and a type VI secretion system (T6SS). The reduced expression of genes involved in T6SS correlated with the detection of the T6SS-effector protein, AssC, in culture supernatants of the A. baumannii wild-type strain, but not in the LPS-deficient strain. Taken together, these data show that, in response to total LPS loss, A. baumannii alters the expression of critical transport and biosynthesis systems associated with modulating the composition and structure of the bacterial surface. Comparison of a gene expression in biological duplicate samples derived from parent bacterial strain to an isogenic mutant strain.

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