Providencia stuartii genes activated by cell-to-cell signaling and identification of a gene required for production or activity of an extracellular factor.
By utilizing reporter transposons, five Providencia stuartii genes that are activated by the accumulation of self-produced extracellular signals have been identified. These genes have been designated cma for conditioned medium activated. The presence of conditioned medium from stationary-phase cultures grown in rich media resulted in the premature activation of each gene in cells at early log phase, with activation values ranging from 6- to 26-fold. Preparation of conditioned medium from an M9 salts medium and fractionation by gel filtration chromatography resulted in fractions within the included volume which activated three of the cma fusions. In addition, depending on the reporter fusion, peak activity was found in different fractions. The partially purified factors activated in a dose-dependent manner. Characterization of the factors activating the cma fusions indicated that they were stable to heat, alkali, and acid. Furthermore, for each cma fusion, factor activity was not reproduced by the addition of homoserine lactone, homocysteine thiolactone, pyruvate, Casamino Acids, or alpha-ketoglutarate. The identities of three cma genes have been determined and revealed physiological roles in amino acid biosynthesis and nutrient import. To begin to address the pathways for production of or response to the extracellular factors, we have identified a locus, aarA, that is required for the activation of four cma fusions. The AarA product was required for factor activity in extracellular supernatants, indicating a possible role in biosynthesis or export.
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Departments of Medicine and Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA. pxr17@po.cwru.edu
Ding
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Baca-DeLancey
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Siddiqui
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D000596:Amino Acids / Q000096:biosynthesis; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002452:Cell Count; D017077:Culture Media, Conditioned / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007763:Lac Operon / Q000235:genetics; D016254:Mutagenesis, Insertional; D011532:Providencia / Q000235:genetics* / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014233:Tricarboxylic Acids / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D000596:Amino Acids; D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014233:Tricarboxylic Acids; C088713:aarA protein, Providencia stuartii; D001616:beta-Galactosidase