20 results on '"Whiteley M"'
Search Results
2. Plasmids of incompatibility group P code for the capacity to propagate bacteriophage IKe
- Author
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Grant, R B, Whiteley, M H, and Shapley, A J
- Abstract
Seven of eight plasmids of incompatibility group P were found to code for the capacity to propagate bacteriophage IKe in Escherichia coli. Six of the seven plasmids allowed propagation of IKe by one bacterial host (RG172) but not by another (RG176); the other plasmid allowed IKe propagation by both hosts. IKe propagation by a number of E. coli K-12 strains was quite variable. IKeh, an extended host range mutant of IKe, was found to plaque specifically on N+ and P+ strains.
- Published
- 1978
- Full Text
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3. Iron-Mediated Control of Pseudomonas aeruginosa-Staphylococcus aureus Interactions in the Cystic Fibrosis Lung.
- Author
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Barnabie PM and Whiteley M
- Subjects
- Humans, Anti-Infective Agents metabolism, Bacteriological Techniques, Coculture Techniques, Cystic Fibrosis microbiology, Iron pharmacology, Microbial Viability, Pseudomonas aeruginosa drug effects, Pseudomonas aeruginosa metabolism, Pseudomonas aeruginosa physiology, Staphylococcus aureus metabolism
- Abstract
Communication is an important factor for bacterial survival, growth, and persistence. Much work has examined both inter- and intraspecies interactions and their effects on virulence. Now, researchers have begun to explore the ways in which host-modulated factors can impact bacterial interactions and subsequently affect patient outcomes. In this issue, two papers discuss how the host environment alters interactions between the pathogens Pseudomonas aeruginosa and Staphylococcus aureus, largely in the context of cystic fibrosis., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
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4. Probing bacterial metabolism during infection using high-resolution transcriptomics.
- Author
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Jorth P, Trivedi U, Rumbaugh K, and Whiteley M
- Subjects
- Animals, Disease Models, Animal, Mice, RNA, Untranslated genetics, RNA, Untranslated isolation & purification, Sequence Analysis, RNA, Transcription Initiation Site, Abscess microbiology, Gene Expression Profiling methods, Metabolic Networks and Pathways genetics, Pasteurellaceae genetics, Pasteurellaceae metabolism, Pasteurellaceae Infections microbiology, Transcriptome
- Abstract
A fundamental aspect of most infectious diseases is the need for the invading microbe to proliferate in the host. However, little is known about the metabolic pathways required for pathogenic microbes to colonize and persist in their hosts. In this study, we used RNA sequencing (RNA-seq) to generate a high-resolution transcriptome of the opportunistic pathogen Aggregatibacter actinomycetemcomitans in vivo. We identified 691 A. actinomycetemcomitans transcriptional start sites and 210 noncoding RNAs during growth in vivo and as a biofilm in vitro. Compared to in vitro biofilm growth on a defined medium, ∼14% of the A. actinomycetemcomitans genes were differentially regulated in vivo. A disproportionate number of genes coding for proteins involved in metabolic pathways were differentially regulated in vivo, suggesting that A. actinomycetemcomitans in vivo metabolism is distinct from in vitro growth. Mutational analyses of differentially regulated genes revealed that formate dehydrogenase H and fumarate reductase are important A. actinomycetemcomitans fitness determinants in vivo. These results not only provide a high-resolution genomic analysis of a bacterial pathogen during in vivo growth but also provide new insight into metabolic pathways required for A. actinomycetemcomitans in vivo fitness.
- Published
- 2013
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5. Role of Pseudomonas aeruginosa peptidoglycan-associated outer membrane proteins in vesicle formation.
- Author
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Wessel AK, Liew J, Kwon T, Marcotte EM, and Whiteley M
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- Protein Binding, Bacterial Proteins metabolism, Exosomes metabolism, Lipoproteins metabolism, Peptidoglycan metabolism, Pseudomonas aeruginosa metabolism
- Abstract
Gram-negative bacteria produce outer membrane vesicles (OMVs) that package and deliver proteins, small molecules, and DNA to prokaryotic and eukaryotic cells. The molecular details of OMV biogenesis have not been fully elucidated, but peptidoglycan-associated outer membrane proteins that tether the outer membrane to the underlying peptidoglycan have been shown to be critical for OMV formation in multiple Enterobacteriaceae. In this study, we demonstrate that the peptidoglycan-associated outer membrane proteins OprF and OprI, but not OprL, impact production of OMVs by the opportunistic pathogen Pseudomonas aeruginosa. Interestingly, OprF does not appear to be important for tethering the outer membrane to peptidoglycan but instead impacts OMV formation through modulation of the levels of the Pseudomonas quinolone signal (PQS), a quorum signal previously shown by our laboratory to be critical for OMV formation. Thus, the mechanism by which OprF impacts OMV formation is distinct from that for other peptidoglycan-associated outer membrane proteins, including OprI.
- Published
- 2013
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6. Pseudomonas aeruginosa enhances production of an antimicrobial in response to N-acetylglucosamine and peptidoglycan.
- Author
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Korgaonkar AK and Whiteley M
- Subjects
- Bacterial Proteins genetics, Bacterial Proteins metabolism, Base Sequence, Molecular Sequence Data, Operon, Promoter Regions, Genetic, Pseudomonas aeruginosa genetics, Acetylglucosamine metabolism, Anti-Bacterial Agents metabolism, Gene Expression Regulation, Bacterial, Peptidoglycan metabolism, Pseudomonas aeruginosa metabolism
- Abstract
Pseudomonas aeruginosa is an opportunistic pathogen often associated with chronic lung infections in individuals with the genetic disease cystic fibrosis (CF). Previous work from our laboratory revealed that five genes predicted to be important for catabolism of N-acetylglucosamine (GlcNAc) are induced during in vitro growth in CF lung secretions (sputum). Here, we demonstrate that these genes comprise an operon (referred to as the nag operon) and that NagE, a putative component of the GlcNAc phosphotransferase system, is required for growth on and uptake of GlcNAc. Using primer extension analysis, the promoter of the nag operon was mapped and shown to be inducible by GlcNAc and regulated by the transcriptional regulator NagR. Transcriptome analysis revealed that in addition to induction of the nag operon, several P. aeruginosa genes encoding factors critical for extracellular antimicrobial production are also induced by GlcNAc. Finally, we show that the GlcNAc-containing polymer peptidoglycan induces production of the antimicrobial pyocyanin. Based on this data, we propose a model in which P. aeruginosa senses surrounding bacteria by monitoring exogenous peptidoglycan and responds to this cue through enhanced production of an antimicrobial.
- Published
- 2011
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7. Characterization of a novel riboswitch-regulated lysine transporter in Aggregatibacter actinomycetemcomitans.
- Author
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Jorth P and Whiteley M
- Subjects
- Computational Biology, Haemophilus influenzae genetics, Membrane Transport Proteins genetics, Gene Expression Regulation, Bacterial, Lysine metabolism, Membrane Transport Proteins biosynthesis, Pasteurellaceae enzymology, Pasteurellaceae genetics, Riboswitch
- Abstract
Aggregatibacter actinomycetemcomitans is an opportunistic pathogen that resides primarily in the mammalian oral cavity. In this environment, A. actinomycetemcomitans faces numerous host- and microbe-derived stresses, including intense competition for nutrients and exposure to the host immune system. While it is clear that A. actinomycetemcomitans responds to precise cues that allow it to adapt and proliferate in the presence of these stresses, little is currently known about the regulatory mechanisms that underlie these responses. Many bacteria use noncoding regulatory RNAs (ncRNAs) to rapidly alter gene expression in response to environmental stresses. Although no ncRNAs have been reported in A. actinomycetemcomitans, we propose that they are likely important for colonization and persistence in the oral cavity. Using a bioinformatic and experimental approach, we identified three putative metabolite-sensing riboswitches and nine small regulatory RNAs (sRNAs) in A. actinomycetemcomitans during planktonic and biofilm growth. Molecular characterization of one of the riboswitches revealed that it is a lysine riboswitch and that its target gene, lysT, encodes a novel lysine-specific transporter. Finally, we demonstrated that lysT and the lysT lysine riboswitch are conserved in over 40 bacterial species, including the phylogenetically related pathogen Haemophilus influenzae.
- Published
- 2010
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8. Characterization of the Pseudomonas aeruginosa transcriptional response to phenylalanine and tyrosine.
- Author
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Palmer GC, Palmer KL, Jorth PA, and Whiteley M
- Subjects
- Bacterial Proteins genetics, Base Sequence, Electrophoretic Mobility Shift Assay, Gene Expression Profiling, Gene Expression Regulation, Bacterial drug effects, Gene Expression Regulation, Bacterial genetics, Genetic Complementation Test, Molecular Sequence Data, Oligonucleotide Array Sequence Analysis, Promoter Regions, Genetic, Protein Binding genetics, Pseudomonas aeruginosa drug effects, Pseudomonas aeruginosa metabolism, Trans-Activators genetics, Bacterial Proteins metabolism, Phenylalanine pharmacology, Pseudomonas aeruginosa genetics, Trans-Activators metabolism, Tyrosine pharmacology
- Abstract
Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen often associated with chronic infections in the lungs of individuals with the heritable disease cystic fibrosis (CF). Previous work from our laboratory demonstrated that aromatic amino acids within CF lung secretions (sputum) not only serve as carbon and energy sources but also enhance synthesis of the cell signaling molecule Pseudomonas quinolone signal (PQS). The present study investigates the role of the aromatic amino acid-responsive regulator PhhR in mediating these phenotypes. Transcriptome analysis revealed that PhhR controls four putative transcriptional units (phhA, hpd, hmgA, and dhcA) involved in aromatic amino acid catabolism; however, genes involved in PQS biosynthesis were unaffected. The phhA, hpd, hmgA, and dhcA promoters were mapped by primer extension, and purified His(6)-PhhR was shown to bind the phhA, hpd, and dhcA promoters in vitro by use of electrophoretic mobility shift assays. Our work characterizes a transcriptional regulator of catabolic genes induced during P. aeruginosa growth in CF sputum.
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- 2010
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9. Characterization of alanine catabolism in Pseudomonas aeruginosa and its importance for proliferation in vivo.
- Author
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Boulette ML, Baynham PJ, Jorth PA, Kukavica-Ibrulj I, Longoria A, Barrera K, Levesque RC, and Whiteley M
- Subjects
- Animals, Bacterial Proteins metabolism, Cell Proliferation, Gene Expression Regulation, Bacterial physiology, Rats, Transcription, Genetic, Alanine metabolism, Peritonitis microbiology, Pseudomonas Infections microbiology, Pseudomonas aeruginosa cytology, Pseudomonas aeruginosa metabolism
- Abstract
The opportunistic pathogen Pseudomonas aeruginosa causes a variety of infections in immunocompromised individuals, including individuals with the heritable disease cystic fibrosis. Like the carbon sources metabolized by many disease-causing bacteria, the carbon sources metabolized by P. aeruginosa at the host infection site are unknown. We recently reported that l-alanine is a preferred carbon source for P. aeruginosa and that two genes potentially involved in alanine catabolism (dadA and dadX) are induced during in vivo growth in the rat peritoneum and during in vitro growth in sputum (mucus) collected from the lungs of individuals with cystic fibrosis. The goals of this study were to characterize factors required for alanine catabolism in P. aeruginosa and to assess the importance of these factors for in vivo growth. Our results reveal that dadA and dadX are arranged in an operon and are required for catabolism of l-alanine. The dad operon is inducible by l-alanine, d-alanine, and l-valine, and induction is dependent on the transcriptional regulator Lrp. Finally, we show that a mutant unable to catabolize dl-alanine displays decreased competitiveness in a rat lung model of infection.
- Published
- 2009
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10. Structural requirements of the Pseudomonas quinolone signal for membrane vesicle stimulation.
- Author
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Mashburn-Warren L, Howe J, Brandenburg K, and Whiteley M
- Subjects
- Lipopolysaccharides chemistry, Lipopolysaccharides metabolism, Models, Molecular, Signal Transduction, Structure-Activity Relationship, Pseudomonas aeruginosa metabolism, Quinolones chemistry, Quinolones metabolism, Secretory Vesicles metabolism
- Abstract
Pseudomonas aeruginosa produces the quorum signal 2-heptyl-3-hydroxy-4-quinolone (Pseudomonas quinolone signal), which is important for stimulating outer membrane vesicle (MV) formation. Here we describe the importance of the 3-hydroxyl and 2-alkyl chain for MV production and the length of the 2-alkyl chain for association with MVs.
- Published
- 2009
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11. Nutritional cues control Pseudomonas aeruginosa multicellular behavior in cystic fibrosis sputum.
- Author
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Palmer KL, Aye LM, and Whiteley M
- Subjects
- Adult, Anti-Bacterial Agents pharmacology, Bacterial Proteins genetics, Bacterial Proteins metabolism, Bacteriological Techniques, Culture Media chemistry, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Humans, Pseudomonas aeruginosa drug effects, Sputum chemistry, Staphylococcus aureus drug effects, Cystic Fibrosis metabolism, Cystic Fibrosis microbiology, Pseudomonas aeruginosa physiology, Sputum metabolism, Sputum microbiology
- Abstract
The sputum (mucus) layer of the cystic fibrosis (CF) lung is a complex substrate that provides Pseudomonas aeruginosa with carbon and energy to support high-density growth during chronic colonization. Unfortunately, the CF lung sputum layer has been difficult to mimic in animal models of CF disease, and mechanistic studies of P. aeruginosa physiology during growth in CF sputum are hampered by its complexity. In this study, we performed chromatographic and enzymatic analyses of CF sputum to develop a defined, synthetic CF sputum medium (SCFM) that mimics the nutritional composition of CF sputum. Importantly, P. aeruginosa displays similar phenotypes during growth in CF sputum and in SCFM, including similar growth rates, gene expression profiles, carbon substrate preferences, and cell-cell signaling profiles. Using SCFM, we provide evidence that aromatic amino acids serve as nutritional cues that influence cell-cell signaling and antimicrobial activity of P. aeruginosa during growth in CF sputum.
- Published
- 2007
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12. A novel exclusion mechanism for carbon resource partitioning in Aggregatibacter actinomycetemcomitans.
- Author
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Brown SA and Whiteley M
- Subjects
- Biological Transport, Models, Biological, Streptococcus metabolism, Fructose metabolism, Glucose metabolism, Pasteurellaceae metabolism, Sodium Lactate metabolism
- Abstract
The bacterium Aggregatibacter actinomycetemcomitans is a common commensal of the human oral cavity and the putative causative agent of the disease localized aggressive periodontitis. A. actinomycetemcomitans is a slow-growing bacterium that possesses limited metabolic machinery for carbon utilization. This likely impacts its ability to colonize the oral cavity, where growth and community composition is mediated by carbon availability. We present evidence that in the presence of the in vivo relevant carbon substrates glucose, fructose, and lactate A. actinomycetemcomitans preferentially metabolizes lactate. This preference for lactate exists despite the fact that A. actinomycetemcomitans grows faster and obtains higher cell yields during growth with carbohydrates. The preference for lactate is mediated by a novel exclusion mechanism in which metabolism of lactate inhibits carbohydrate uptake. Coculture studies reveal that A. actinomycetemcomitans utilizes lactate produced by the oral bacterium Streptococcus gordonii, suggesting the potential for cross-feeding in the oral cavity.
- Published
- 2007
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13. Membrane-bound nitrate reductase is required for anaerobic growth in cystic fibrosis sputum.
- Author
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Palmer KL, Brown SA, and Whiteley M
- Subjects
- Anaerobiosis, Artificial Gene Fusion, Bacterial Proteins genetics, Base Sequence, Colony Count, Microbial, Gene Deletion, Gene Expression Regulation, Bacterial, Genes, Reporter, Membrane Proteins genetics, Microbial Viability, Molecular Sequence Data, Mutagenesis, Insertional, Nitrate Reductase biosynthesis, Nitrate Reductase genetics, Nitrates metabolism, Sputum chemistry, Transcription, Genetic, beta-Galactosidase biosynthesis, beta-Galactosidase genetics, Bacterial Proteins physiology, Cystic Fibrosis microbiology, Membrane Proteins physiology, Nitrate Reductase physiology, Pseudomonas aeruginosa enzymology, Pseudomonas aeruginosa growth & development, Sputum microbiology
- Abstract
The autosomal recessive disorder cystic fibrosis (CF) affects approximately 70,000 people worldwide and is characterized by chronic bacterial lung infections with the opportunistic pathogen Pseudomonas aeruginosa. To form a chronic CF lung infection, P. aeruginosa must grow and proliferate within the CF lung, and the highly viscous sputum within the CF lung provides a likely growth substrate. Recent evidence indicates that anaerobic microenvironments may be present in the CF lung sputum layer. Since anaerobic growth significantly enhances P. aeruginosa biofilm formation and antibiotic resistance, it is important to examine P. aeruginosa physiology and metabolism in anaerobic environments. Measurement of nitrate levels revealed that CF sputum contains sufficient nitrate to support significant P. aeruginosa growth anaerobically, and mutational analysis revealed that the membrane-bound nitrate reductase is essential for P. aeruginosa anaerobic growth in an in vitro CF sputum medium. In addition, expression of genes coding for the membrane-bound nitrate reductase complex is responsive to CF sputum nitrate levels. These findings suggest that the membrane-bound nitrate reductase is critical for P. aeruginosa anaerobic growth with nitrate in the CF lung.
- Published
- 2007
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14. Survival and growth in the presence of elevated copper: transcriptional profiling of copper-stressed Pseudomonas aeruginosa.
- Author
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Teitzel GM, Geddie A, De Long SK, Kirisits MJ, Whiteley M, and Parsek MR
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- Adaptation, Physiological, Cell Membrane Permeability, Copper metabolism, Homeostasis, Iron metabolism, Oligonucleotide Array Sequence Analysis, Pseudomonas aeruginosa growth & development, Pseudomonas aeruginosa metabolism, RNA, Bacterial analysis, RNA, Bacterial genetics, RNA, Messenger analysis, RNA, Messenger genetics, Transcription, Genetic, Zinc toxicity, Copper toxicity, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Microbial Viability, Pseudomonas aeruginosa drug effects, Pseudomonas aeruginosa genetics
- Abstract
Transcriptional profiles of Pseudomonas aeruginosa exposed to two separate copper stress conditions were determined. Actively growing bacteria subjected to a pulse of elevated copper for a short period of time was defined as a "copper-shocked" culture. Conversely, copper-adapted populations were defined as cells actively growing in the presence of elevated copper. Expression of 405 genes changed in the copper-shocked culture, compared to 331 genes for the copper-adapted cultures. Not surprisingly, there were genes identified in common to both conditions. For example, both stress conditions resulted in up-regulation of genes encoding several active transport functions. However, there were some interesting differences between the two types of stress. Only copper-adapted cells significantly altered expression of passive transport functions, down-regulating expression of several porins belonging to the OprD family. Copper shock produced expression profiles suggestive of an oxidative stress response, probably due to the participation of copper in Fenton-like chemistry. Copper-adapted populations did not show such a response. Transcriptional profiles also indicated that iron acquisition is fine-tuned in the presence of copper. Several genes induced under iron-limiting conditions, such as the siderophore pyoverdine, were up-regulated in copper-adapted populations. Interesting exceptions were the genes involved in the production of the siderophore pyochelin, which were down-regulated. Analysis of the copper sensitivity of select mutant strains confirmed the array data. These studies suggest that two resistance nodulation division efflux systems, a P-type ATPase, and a two-component regulator were particularly important for copper tolerance in P. aeruginosa.
- Published
- 2006
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15. Bundle-forming pili and EspA are involved in biofilm formation by enteropathogenic Escherichia coli.
- Author
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Moreira CG, Palmer K, Whiteley M, Sircili MP, Trabulsi LR, Castro AF, and Sperandio V
- Subjects
- Genotype, Kinetics, Mutagenesis, Recombinant Proteins metabolism, Signal Transduction, Transcription, Genetic, Biofilms growth & development, Escherichia coli pathogenicity, Escherichia coli physiology, Escherichia coli Proteins genetics, Escherichia coli Proteins metabolism, Fimbriae, Bacterial physiology
- Abstract
Microcolony formation is one of the initial steps in biofilm development, and in enteropathogenic Escherichia coli (EPEC) it is mediated by several adhesins, including the bundle-forming pilus (BFP) and the EspA filament. Here we report that EPEC forms biofilms on plastic under static conditions and a flowthrough continuous culture system. The abilities of several EPEC isogenic mutants to form biofilms were assessed. Adhesins such as BFP and EspA, important in microcolony formation on epithelial cells, are also involved in bacterial aggregation during biofilm formation on abiotic surfaces. Mutants that do not express BFP or EspA form more-diffuse biofilms than does the wild type. We also determined, using gfp transcriptional fusions, that, consistent with the role of these adhesins in biofilms, the genes encoding BFP and EspA are expressed during biofilm formation. Finally, expression of espA is controlled by a quorum-sensing (QS) regulatory mechanism, and the EPEC qseA QS mutant also forms altered biofilms, suggesting that this signaling mechanism plays an important role in EPEC biofilm development. Taken together, these studies allowed us to propose a model of EPEC biofilm formation.
- Published
- 2006
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16. Microarray analysis of the osmotic stress response in Pseudomonas aeruginosa.
- Author
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Aspedon A, Palmer K, and Whiteley M
- Subjects
- Betaine, Osmolar Concentration, Time Factors, Bacterial Proteins genetics, Gene Expression Profiling, Oligonucleotide Array Sequence Analysis, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa metabolism
- Abstract
Transcriptional profiling of Pseudomonas aeruginosa grown under steady-state hyperosmotic stress conditions showed an up-regulation of genes associated with osmoprotectant synthesis, putative hydrophilins, and the type III secretion system with associated cytotoxins. A large number of regulatory genes, including several two-component systems not previously known to be influenced by osmolarity, were differentially expressed by P. aeruginosa in immediate response to hyperosmotic shock.
- Published
- 2006
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17. Cystic fibrosis sputum supports growth and cues key aspects of Pseudomonas aeruginosa physiology.
- Author
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Palmer KL, Mashburn LM, Singh PK, and Whiteley M
- Subjects
- Culture Media, Cystic Fibrosis microbiology, Humans, Locomotion, Pseudomonas aeruginosa growth & development, Sputum, Pseudomonas aeruginosa physiology
- Abstract
The opportunistic human pathogen Pseudomonas aeruginosa causes persistent airway infections in patients with cystic fibrosis (CF). To establish these chronic infections, P. aeruginosa must grow and proliferate within the highly viscous sputum in the lungs of CF patients. In this study, we used Affymetrix GeneChip microarrays to investigate the physiology of P. aeruginosa grown using CF sputum as the sole source of carbon and energy. Our results indicate that CF sputum readily supports high-density P. aeruginosa growth. Furthermore, multiple signals, which reduce swimming motility and prematurely activate the Pseudomonas quinolone signal cell-to-cell signaling cascade in P. aeruginosa, are present in CF sputum. P. aeruginosa factors critical for lysis of the common CF lung inhabitant Staphylococcus aureus were also induced in CF sputum and increased the competitiveness of P. aeruginosa during polymicrobial growth in CF sputum.
- Published
- 2005
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18. Staphylococcus aureus serves as an iron source for Pseudomonas aeruginosa during in vivo coculture.
- Author
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Mashburn LM, Jett AM, Akins DR, and Whiteley M
- Subjects
- Animals, Antibiosis, Bacterial Proteins genetics, Culture Media chemistry, Disease Models, Animal, Genes, Bacterial, Peritoneal Cavity microbiology, Peritoneal Dialysis, Pseudomonas aeruginosa metabolism, Rats, Staphylococcus aureus metabolism, Gene Expression Profiling, Gene Expression Regulation, Bacterial, Iron metabolism, Oligonucleotide Array Sequence Analysis, Pseudomonas aeruginosa growth & development, Staphylococcus aureus growth & development
- Abstract
Pseudomonas aeruginosa is a gram-negative opportunistic human pathogen often infecting the lungs of individuals with the heritable disease cystic fibrosis and the peritoneum of individuals undergoing continuous ambulatory peritoneal dialysis. Often these infections are not caused by colonization with P. aeruginosa alone but instead by a consortium of pathogenic bacteria. Little is known about growth and persistence of P. aeruginosa in vivo, and less is known about the impact of coinfecting bacteria on P. aeruginosa pathogenesis and physiology. In this study, a rat dialysis membrane peritoneal model was used to evaluate the in vivo transcriptome of P. aeruginosa in monoculture and in coculture with Staphylococcus aureus. Monoculture results indicate that approximately 5% of all P. aeruginosa genes are differentially regulated during growth in vivo compared to in vitro controls. Included in this analysis are genes important for iron acquisition and growth in low-oxygen environments. The presence of S. aureus caused decreased transcription of P. aeruginosa iron-regulated genes during in vivo coculture, indicating that the presence of S. aureus increases usable iron for P. aeruginosa in this environment. We propose a model where P. aeruginosa lyses S. aureus and uses released iron for growth in low-iron environments.
- Published
- 2005
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19. Promoter specificity elements in Pseudomonas aeruginosa quorum-sensing-controlled genes.
- Author
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Whiteley M and Greenberg EP
- Subjects
- Bacterial Proteins metabolism, Base Sequence, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Gene Deletion, Molecular Sequence Data, Point Mutation, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa metabolism, Signal Transduction, Trans-Activators genetics, Trans-Activators metabolism, Bacterial Proteins genetics, Gene Expression Regulation, Bacterial, Promoter Regions, Genetic genetics, Pseudomonas aeruginosa growth & development, Transcription, Genetic
- Abstract
The LasR-dependent and RhlR-dependent quorum-sensing systems are global regulators of gene expression in Pseudomonas aeruginosa. Previous studies have demonstrated that promoter elements of the quorum-sensing-controlled genes lasB and hcnABC are important in density-dependent regulation. We have identified LasR- and RhlR-dependent determinants in promoters of quorum-sensing-controlled genes qsc102, qsc117 (acpP), and qsc131 (phzA to -G) by in silico, deletion, point-mutational, and primer extension analyses. Each of these genes (in addition to lasI and rsaL) is activated by LasR, and qsc117 and qsc131 also respond to RhlR. Point mutations in the promoters of the LasR-specific gene, qsc102, relax specificity so that this promoter can respond to RhlR in addition to LasR. Our findings indicate that quorum-sensing-controlled promoters in P. aeruginosa are either specific for LasR or respond to both LasR and RhlR and that critical bases in the promoter elements determine specificity.
- Published
- 2001
- Full Text
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20. Regulation of quorum sensing by RpoS in Pseudomonas aeruginosa.
- Author
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Whiteley M, Parsek MR, and Greenberg EP
- Subjects
- Colony Count, Microbial, Ligases, Pseudomonas aeruginosa pathogenicity, Pyocyanine metabolism, Transcription Factors physiology, Transcription, Genetic, Bacterial Proteins physiology, Pseudomonas aeruginosa genetics, Sigma Factor physiology
- Abstract
The LasR-LasI and RhlR-RhlI quorum-sensing systems are global regulators of gene expression in the opportunistic pathogen Pseudomonas aeruginosa. Previous studies suggest that the RhlR-RhlI system activates expression of rpoS. We constructed merodiploid strains of P. aeruginosa containing the native rpoS gene and an rpoS-lacZ fusion. Studies of lacZ transcription in these strains indicated that rpoS was not regulated by RhlR-RhlI. We also generated an rpoS null mutant. This rpoS mutant showed elevated levels of rhlI (but not rhlR) transcription, elevated levels of the RhlI-generated acylhomoserine lactone quorum-sensing signal, and elevated levels of RhlR-RhlI-regulated gene transcription. These findings indicate that there is a relationship between RpoS and quorum sensing, but rather than the RhlR-RhlI system influencing the expression of rpoS, it appears that RpoS regulates rhlI.
- Published
- 2000
- Full Text
- View/download PDF
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