An ordered, nonredundant library of
            <i>Pseudomonas aeruginosa</i>
            strain PA14 transposon insertion mutants

Liberati, Nicole T.; Urbach, Jonathan M.; Miyata, Sachiko; Lee, Daniel G.; Drenkard, Eliana; Wu, Gang; Villanueva, Jacinto M.; Wei, Tao; Ausubel, Frederick M.

doi:10.1073/pnas.0511100103

Cited by 907 publications

(982 citation statements)

References 40 publications

Supporting

Mentioning

942

Contrasting

Unclassified

Order By: Relevance

“…using a Big Dye fluorescent terminator and an ABI3770 capillary sequencer at the Dana Farber Cancer Center Core Laboratory. Transposon mutagenesis was performed as previously described with a mariner element derivative of Mar2xT7 (Liberati et al 2006).…”

Section: Molecular and Genetic Techniquesmentioning

confidence: 99%

Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen

Goodman¹,

Merighi²,

Hyodo³

et al. 2009

Genes Dev.

275

297

View full text Add to dashboard Cite

The genome of the opportunistic pathogen Pseudomonas aeruginosa encodes over 60 two-component sensor kinases and uses several (including RetS and GacS) to reciprocally regulate the production of virulence factors involved in the development of acute or chronic infections. We demonstrate that RetS modulates the phosphorylation state of GacS by a direct and specific interaction between these two membrane-bound sensors. The RetS-GacS interaction can be observed in vitro, in heterologous systems in vivo, and in P. aeruginosa. This function does not require the predicted RetS phosphorelay residues and provides a mechanism for integrating multiple signals without cross-phosphorylation from sensors to noncognate response regulators. These results suggest that multiple two-component systems found in a single bacterium can form multisensor signaling networks while maintaining specific phosphorelay pathways that remain insulated from detrimental cross-talk. Two-component system (TCS) signaling pathways are a major signaling mechanism in bacteria and archaea, and are also found in simple eukaryota and higher plants (Wolanin et al. 2002). These diverse organisms capitalize on TCS pathways to monitor critical external and internal stimuli (including levels of nutrients, concentration of ions and gases, temperature, redox states, and cell density) and translate these signals into adaptive responses. Classical TCS pathways share a conserved core architecture: a homodimerizing histidine kinase protein domain (the ''sensor'') and a cognate receiver domain (the ''response regulator''), coupled mechanistically through a histidine-aspartic acid phosphorelay (Stock et al. 2000). Most cognate sensor response regulator pairs are also linked genetically, encoded by adjacent loci in the chromosome (Alm et al. 2006). Although a single bacterial species can encode up to hundreds of genes specifying TCS pathways, it appears that these systems are insulated against detrimental cross-phosphorylation between sensors and noncognate response regulators (Bijlsma and Groisman 2003;Baker and Stock 2007;Laub and Goulian 2007). The identification of multistep phosphorelays (with intermediary proteins between sensor and regulator) and branched pathways (phosphotransfer between one sensor and multiple response regulators and vice versa) (Laub and Goulian 2007) suggests that TCS pathways have the capacity to form sensitive and complex signaling networks.In contrast to microorganisms with restricted habitats, the genomes of bacteria capable of occupying a number of diverse environments typically contain a disproportionately large number of genes encoding signal transduction and regulatory systems, including TCSs that allow them to sense and respond to a wide range of environmental signals. For opportunistic bacterial pathogens, a number of these systems regulate the expression of genes necessary for transitioning from the environmental reservoir to the host, overcoming innate defense mechanisms and initiating the disease process. The bacterial pathogen ...

show abstract

Section: Molecular and Genetic Techniquesmentioning

confidence: 99%

Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen

Goodman¹,

Merighi²,

Hyodo³

et al. 2009

Genes Dev.

275

297

View full text Add to dashboard Cite

show abstract

“…Wild-type P. aeruginosa PA14 was used instead of PAO1 because PA14 is more virulent than PAO1 in diverse infection models (Harrison et al, 2010) and because of the availability of the complete mutant library (Liberati et al, 2006). Our strategy was to utilize a minimal medium (so it resembles more closely clinical situations) using a growth compound whose assimilation requires QS; therefore, QS and growth were inhibited by the antivirulence compound C-30, which has become the gold standard for antivirulence compounds.…”

Section: Introductionmentioning

confidence: 99%

Quorum quenching quandary: resistance to antivirulence compounds

et al. 2011

View full text Add to dashboard Cite

Quorum sensing (QS) is the regulation of gene expression in response to the concentration of small signal molecules, and its inactivation has been suggested to have great potential to attenuate microbial virulence. It is assumed that unlike antimicrobials, inhibition of QS should cause less Darwinian selection pressure for bacterial resistance. Using the opportunistic pathogen Pseudomonas aeruginosa, we demonstrate here that bacterial resistance arises rapidly to the best-characterized compound that inhibits QS (brominated furanone C-30) due to mutations that increase the efflux of C-30. Critically, the C-30-resistant mutant mexR was more pathogenic to Caenorhabditis elegans in the presence of C-30, and the same mutation arises in bacteria responsible for chronic cystic fibrosis infections. Therefore, bacteria may evolve resistance to many new pharmaceuticals thought impervious to resistance.

show abstract

“…This is true both for the mutants we generated and for PA14 strains that have a gentamicin resistance cassette (aacC1) inserted into their genome [15]. These data indicate that growing cells also inhibit antibiotic-resistant mutants, and that mutants can inhibit their clonal siblings.…”

Section: The Inhibitory Factor Is Produced Without Antibiotic But Caumentioning

confidence: 58%

The spatial profiles and metabolic capabilities of microbial populations impact the growth of antibiotic-resistant mutants

Kaushik

Ratnayeke

Katira

et al. 2015

J. R. Soc. Interface.

View full text Add to dashboard Cite

Antibiotic resistance adversely affects clinical and public health on a global scale. Using the opportunistic human pathogen Pseudomonas aeruginosa, we show that increasing the number density of bacteria, on agar containing aminoglycoside antibiotics, can non-monotonically impact the survival of antibiotic-resistant mutants. Notably, at high cell densities, mutant survival is inhibited. A wide range of bacterial species can inhibit antibiotic-resistant mutants. Inhibition results from the metabolic breakdown of amino acids, which results in alkaline by-products. The consequent increase in pH acts in conjunction with aminoglycosides to mediate inhibition. Our work raises the possibility that the manipulation of microbial population structure and nutrient environment in conjunction with existing antibiotics could provide therapeutic approaches to combat antibiotic resistance.

show abstract

An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants

Cited by 907 publications

References 40 publications

Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen

Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen

Quorum quenching quandary: resistance to antivirulence compounds

The spatial profiles and metabolic capabilities of microbial populations impact the growth of antibiotic-resistant mutants

Contact Info

Product

Resources

About