title: SpoT-triggered stringent response controls usp gene expression in Pseudomonas aeruginosa.
abstract: The universal stress proteins (Usps) UspK (PA3309) and UspN (PA4352) of Pseudomonas aeruginosa are essential for surviving specific anaerobic energy stress conditions such as pyruvate fermentation and anaerobic stationary phase. Expression of the respective genes is under the control of the oxygen-sensing regulator Anr. In this study we investigated the regulation of uspN and three additional P. aeruginosa usp genes: uspL (PA1789), uspM (PA4328), and uspO (PA5027). Anr induces expression of these genes in response to anaerobic conditions. Using promoter-lacZ fusions, we showed that PuspL-lacZ, PuspM-lacZ, and PuspO-lacZ were also induced in stationary phase as described for PuspN-lacZ. However, stationary phase gene expression was abolished in the P. aeruginosa triple mutant Deltaanr DeltarelA DeltaspoT. The relA and spoT genes encode the regulatory components of the stringent response. We determined pppGpp and ppGpp levels using a thin-layer chromatography approach and detected the accumulation of ppGpp in the wild type and the DeltarelA mutant in stationary phase, indicating a SpoT-derived control of ppGpp accumulation. Additional investigation of stationary phase in LB medium revealed that alkaline pH values are involved in the regulatory process of ppGpp accumulation.
authors.affiliation: Institute of Microbiology, Technische Universität Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany.
authors.forename: Nelli
authors.initials: N
authors.lastname: Boes
authors.forename: Kerstin
authors.initials: K
authors.lastname: Schreiber
authors.forename: Max
authors.initials: M
authors.lastname: Schobert
mesh_terms: D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D002855:Chromatography, Thin Layer; D015964:Gene Expression Regulation, Bacterial; D006158:Guanosine Pentaphosphate / Q000378:metabolism; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D011401:Promoter Regions, Genetic / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D015534:Trans-Activators; C072908:ANR protein, Pseudomonas aeruginosa; D006159:Guanosine Tetraphosphate; D006158:Guanosine Pentaphosphate; D001616:beta-Galactosidase
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