Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 93

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

Moonlighting chaperone activity of the enzyme PqsE contributes to RhlR-controlled virulence of Pseudomonas aeruginosa

Borgert, Sebastian Roman, Henke, Steffi, Witzgall, Florian, Schmelz, Stefan, zur Lage, Susanne, Hotop, Sven-Kevin, Stephen, Steffi, Lübken, Dennis, Krüger, Jonas, Gomez, Nicolas Oswaldo, van Ham, Marco, Jänsch, Lothar, Kalesse, Markus, Pich, Andreas, Brönstrup, Mark, Häussler, Susanne, Blankenfeldt, Wulf·2022·Nature Communications
DOI 10.1038/s41467-022-35030-w

Published version of a preprint

Open this paper in CoChat — free account

Abstract

Pseudomonas aeruginosa is a major cause of nosocomial infections and also leads to severe exacerbations in cystic fibrosis or chronic obstructive pulmonary disease. Three intertwined quorum sensing systems control virulence of P. aeruginosa , with the rhl circuit playing the leading role in late and chronic infections. The majority of traits controlled by rhl transcription factor RhlR depend on PqsE, a dispensable thioesterase in Pseudomonas Quinolone Signal (PQS) biosynthesis that interferes with RhlR through an enigmatic mechanism likely involving direct interaction of both proteins. Here we show that PqsE and RhlR form a 2:2 protein complex that, together with RhlR agonist N -butanoyl-L-homoserine lactone (C4-HSL), solubilizes RhlR and thereby renders the otherwise insoluble transcription factor active. We determine crystal structures of the complex and identify residues essential for the interaction. To corroborate the chaperone-like activity of PqsE, we design stability-optimized variants of RhlR that bypass the need for C4-HSL and PqsE in activating PqsE/RhlR-controlled processes of P. aeruginosa . Together, our data provide insight into the unique regulatory role of PqsE and lay groundwork for developing new P. aeruginosa -specific pharmaceuticals.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/s41467-022-35030-w

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 3 references

Research works printed but not linked: 70

  1. Morin, C. D., Déziel, E., Gauthier, J., Levesque, R. C. & Lau, G. W. An Organ System-Based Synopsis of Pseudomonas aeruginosa Virulence. Virulence 12, 1469–1507 (2021). · DOI
  2. Murray, C. J. et al. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. The Lancet 399, 629–655 (2022). · DOI · Paper page
  3. Diggle, S. P. & Whiteley, M. Microbe Profile: Pseudomonas aeruginosa: opportunistic pathogen and lab rat. Microbiol. Read. Engl. 166, 30–33 (2020). · DOI · Paper page
  4. Dietrich, L. E., Teal, T. K., Price-Whelan, A. & Newman, D. K. Redox-active antibiotics control gene expression and community behavior in divergent bacteria. Science 321, 1203–1206 (2008). · DOI · Paper page
  5. Ahator, S. D. & Zhang, L. Small Is Mighty-Chemical Communication Systems in Pseudomonas aeruginosa. Annu. Rev. Microbiol. 73, 559–578 (2019). · DOI
  6. Diggle, S. P. et al. Functional genetic analysis reveals a 2-Alkyl-4-quinolone signaling system in the human pathogen Burkholderia pseudomallei and related bacteria. Chem. Biol. 13, 701–710 (2006). · doi:10.1016/j.chembiol.2006.05.006
  7. Gallagher, L. A., McKnight, S. L., Kuznetsova, M. S., Pesci, E. C. & Manoil, C. Functions required for extracellular quinolone signaling by Pseudomonas aeruginosa. J. Bacteriol. 184, 6472–6480 (2002). · DOI · Paper page
  8. Pesci, E. C. et al. Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa. Proc. Natl Acad. Sci. 96, 11229–11234 (1999). · DOI · Paper page
  9. Brouwer, S. et al. The PqsR and RhlR transcriptional regulators determine the level of PQS synthesis in Pseudomonas aeruginosa by producing two different pqsABCDE mRNA isoforms. J. Bacteriol. 196, 4163–4171 (2014). · doi:10.1128/jb.02000-14
  10. Dekimpe, V. & Déziel, E. Revisiting the quorum-sensing hierarchy in Pseudomonas aeruginosa: the transcriptional regulator RhlR regulates LasR-specific factors. Microbiology 155, 712–723 (2009). · DOI · Paper page
  11. Feltner, J. B. et al. LasR Variant Cystic Fibrosis Isolates Reveal an Adaptable Quorum-Sensing Hierarchy in Pseudomonas aeruginosa. mBio 7, e01513–e01516 (2016). · DOI · Paper page
  12. Groleau, M.-C., Taillefer, H., Vincent, A. T., Constant, P. & Déziel, E. Pseudomonas aeruginosa isolates defective in function of the LasR quorum sensing regulator are frequent in diverse environmental niches. Environ. Microbiol. 24, 1062–1075 (2021). · DOI
  13. Gilbert, K. B., Kim, T. H., Gupta, R., Greenberg, E. P. & Schuster, M. Global position analysis of the Pseudomonas aeruginosa quorum-sensing transcription factor LasR. Mol. Microbiol. 73, 1072–1085 (2009). · DOI · Paper page
  14. Mukherjee, S., Moustafa, D., Smith, C. D., Goldberg, J. B. & Bassler, B. L. The RhlR quorum-sensing receptor controls Pseudomonas aeruginosa pathogenesis and biofilm development independently of its canonical homoserine lactone autoinducer. PLoS Pathog. 13, e1006504 (2017). · DOI · Paper page
  15. Letizia, M. et al. PqsE Expands and Differentially Modulates the RhlR Quorum Sensing Regulon in Pseudomonas aeruginosa. Microbiol. Spectr. 10, e0096122 (2022). · DOI · Paper page
  16. Rampioni, G. et al. Unravelling the Genome-Wide Contributions of Specific 2-Alkyl-4-Quinolones and PqsE to Quorum Sensing in Pseudomonas aeruginosa. PLoS Pathog. 12, e1006029 (2016). · DOI · Paper page
  17. Xiao, G. et al. MvfR, a key Pseudomonas aeruginosa pathogenicity LTTR-class regulatory protein, has dual ligands. Mol. Microbiol. 62, 1689–1699 (2006). · DOI · Paper page
  18. Yu, S. et al. Structure elucidation and preliminary assessment of hydrolase activity of PqsE, the Pseudomonas quinolone signal (PQS) response protein. Biochemistry 48, 10298–10307 (2009). · DOI
  19. Drees, S. L. & Fetzner, S. PqsE of Pseudomonas aeruginosa Acts as Pathway-Specific Thioesterase in the Biosynthesis of Alkylquinolone Signaling Molecules. Chem. Biol. 22, 611–618 (2015). · DOI
  20. Déziel, E. et al. The contribution of MvfR to Pseudomonas aeruginosa pathogenesis and quorum sensing circuitry regulation: multiple quorum sensing-regulated genes are modulated without affecting lasRI, rhlRI or the production of N-acyl-L-homoserine lactones. Mol. Microbiol. 55, 998–1014 (2005). · DOI · Paper page
  21. Mukherjee, S. et al. The PqsE and RhlR proteins are an autoinducer synthase-receptor pair that control virulence and biofilm development in Pseudomonas aeruginosa. Proc. Natl Acad. Sci. 115, E9411–E9418 (2018). · DOI
  22. Farrow, J. M. et al. PqsE functions independently of PqsR-Pseudomonas quinolone signal and enhances the rhl quorum-sensing system. J. Bacteriol. 190, 7043–7051 (2008). · DOI · Paper page
  23. Tsai, C.-S. & Winans, S. C. LuxR-type quorum-sensing regulators that are detached from common scents. Mol. Microbiol. 77, 1072–1082 (2010). · doi:10.1111/j.1365-2958.2010.07279.x
  24. Churchill, M. E. A. & Chen, L. Structural basis of acyl-homoserine lactone-dependent signaling. Chem. Rev. 111, 68–85 (2011). · DOI · Paper page
  25. O’Loughlin, C. T. et al. A quorum-sensing inhibitor blocks Pseudomonas aeruginosa virulence and biofilm formation. Proc. Natl Acad. Sci. 110, 17981–17986 (2013). · DOI · Paper page
  26. McCready, A. R., Paczkowski, J. E., Cong, J.-P. & Bassler, B. L. An autoinducer-independent RhlR quorum-sensing receptor enables analysis of RhlR regulation. PLoS Pathog. 15, e1007820 (2019). · DOI
  27. Groleau, M.-C., de Pereira, T. O., Dekimpe, V. & Déziel, E. PqsE Is Essential for RhlR-Dependent Quorum Sensing Regulation in Pseudomonas aeruginosa. mSystems 5, e00194-20 (2020). · DOI · Paper page
  28. Taylor, I. R. et al. Inhibitor Mimetic Mutations in the Pseudomonas aeruginosa PqsE Enzyme Reveal a Protein-Protein Interaction with the Quorum-Sensing Receptor RhlR That Is Vital for Virulence Factor Production. ACS Chem. Biol. 16, 740–752 (2021). · DOI · Paper page
  29. Simanek, K. A. et al. The PqsE-RhlR Interaction Regulates RhlR DNA Binding to Control Virulence Factor Production in Pseudomonas aeruginosa. Microbiol. Spectr. 10, e0210821 (2022). · DOI · Paper page
  30. Folch, B., Déziel, E. & Doucet, N. Systematic Mutational Analysis of the Putative Hydrolase PqsE: Toward a Deeper Molecular Understanding of Virulence Acquisition in Pseudomonas aeruginosa. PLOS ONE 8, e73727 (2013). · doi:10.1371/journal.pone.0073727
  31. Zender, M. et al. Dissecting the Multiple Roles of PqsE in Pseudomonas aeruginosa Virulence by Discovery of Small Tool Compounds. ACS Chem. Biol. 11, 1755–1763 (2016). · DOI
  32. Taylor, I. R. et al. Protein Interaction Networks of Catalytically Active and Catalytically Inactive PqsE in Pseudomonas aeruginosa. mBio 13, e01559–22 (2022). · DOI
  33. Cataldi, T. R. I., Bianco, G. & Abate, S. Accurate mass analysis of N-acyl-homoserine-lactones and cognate lactone-opened compounds in bacterial isolates of Pseudomonas aeruginosa PAO1 by LC-ESI-LTQ-FTICR-MS. J. Mass Spectrom. 44, 182–192 (2009). · DOI
  34. Ortori, C. A. et al. Simultaneous quantitative profiling of N-acyl-L-homoserine lactone and 2-alkyl-4(1H)-quinolone families of quorum-sensing signaling molecules using LC-MS/MS. Anal. Bioanal. Chem. 399, 839–850 (2011). · DOI
  35. Henkel, M. et al. Kinetic modeling of the time course of N-butyryl-homoserine lactone concentration during batch cultivations of Pseudomonas aeruginosa PAO1. Appl. Microbiol. Biotechnol. 97, 7607–7616 (2013). · DOI
  36. Pustelny, C. et al. The peptide chain release factor methyltransferase PrmC is essential for pathogenicity and environmental adaptation of Pseudomonas aeruginosa PA14. Environ. Microbiol. 15, 597–609 (2013). · DOI
  37. García-Reyes, S. et al. The PqsR-independent quorum-sensing response of Pseudomonas aeruginosa ATCC 9027 outlier-strain reveals new insights on the PqsE effect on RhlR activity. Mol. Microbiol. 116, 1113–1123 (2021). · DOI
  38. Berman, H. M. et al. The Protein Data Bank. Nucl. Acids Res. 28, 235–242 (2000). · DOI · Paper page
  39. Frank, R. Spot-synthesis: an easy technique for the positionally addressable, parallel chemical synthesis on a membrane support. Tetrahedron 48, 9217–9232 (1992). · DOI · Paper page
  40. Schellenberger, V. et al. A recombinant polypeptide extends the in vivo half-life of peptides and proteins in a tunable manner. Nat. Biotechnol. 27, 1186–1190 (2009). · DOI · Paper page
  41. Bottomley, M. J., Muraglia, E., Bazzo, R. & Carfì, A. Molecular insights into quorum sensing in the human pathogen Pseudomonas aeruginosa from the structure of the virulence regulator LasR bound to its autoinducer. J. Biol. Chem. 282, 13592–13600 (2007). · DOI · Paper page
  42. Goldenzweig, A. et al. Automated Structure- and Sequence-Based Design of Proteins for High Bacterial Expression and Stability. Mol. Cell. 63, 337–346 (2016). · DOI · Paper page
  43. Paczkowski, J. E. et al. An Autoinducer Analogue Reveals an Alternative Mode of Ligand Binding for the LasR Quorum-Sensing Receptor. ACS Chem. Biol. 14, 378–389 (2019). · DOI
  44. Rasko, D. A. & Sperandio, V. Anti-virulence strategies to combat bacteria-mediated disease. Nat. Rev. Drug Discov. 9, 117–128 (2010). · DOI · Paper page
  45. Maura, D. & Rahme, L. G. Pharmacological Inhibition of the Pseudomonas aeruginosa MvfR Quorum-Sensing System Interferes with Biofilm Formation and Potentiates Antibiotic-Mediated Biofilm Disruption. Antimicrob. Agents Chemother. 61, e01362–17 (2017). · doi:10.1128/aac.01362-17
  46. Kitao, T. et al. Molecular Insights into Function and Competitive Inhibition of Pseudomonas aeruginosa Multiple Virulence Factor Regulator. mBio 9, e02158–17 (2018). · DOI
  47. Zender, M. et al. Flexible Fragment Growing Boosts Potency of Quorum-Sensing Inhibitors against Pseudomonas aeruginosa Virulence. ChemMedChem 15, 188–194 (2020). · doi:10.1002/cmdc.201900621
  48. Grossman, S. et al. Novel quinazolinone inhibitors of the Pseudomonas aeruginosa quorum sensing transcriptional regulator PqsR. Eur. J. Med. Chem. 208, 112778 (2020). · DOI
  49. Soukarieh, F. et al. Hit Identification of New Potent PqsR Antagonists as Inhibitors of Quorum Sensing in Planktonic and Biofilm Grown Pseudomonas aeruginosa. Front. Chem. 8, 204 (2020). · DOI
  50. Sabir, S. et al. Thioether-linked dihydropyrrol-2-one analogues as PqsR antagonists against antibiotic resistant Pseudomonas aeruginosa. Bioorg. Med. Chem. 31, 115967 (2021). · doi:10.1016/j.bmc.2020.115967
  51. Schütz, C. et al. A New PqsR Inverse Agonist Potentiates Tobramycin Efficacy to Eradicate Pseudomonas aeruginosa Biofilms. Adv. Sci. 8, e2004369 (2021). · DOI
  52. Dolan, K. M. & Greenberg, E. P. Evidence that GroEL, not sigma 32, is involved in transcriptional regulation of the Vibrio fischeri luminescence genes in Escherichia coli. J. Bacteriol. 174, 5132–5135 (1992). · DOI
  53. Urbanowski, M. L., Lostroh, C. P. & Greenberg, E. P. Reversible Acyl-Homoserine Lactone Binding to Purified Vibrio fischeri LuxR Protein. J. Bacteriol. 186, 631–637 (2004). · doi:10.1128/jb.186.3.631-637.2004
  54. Lee, J.-H., Lequette, Y. & Greenberg, E. P. Activity of purified QscR, a Pseudomonas aeruginosa orphan quorum-sensing transcription factor. Mol. Microbiol. 59, 602–609 (2006). · DOI · Paper page
  55. Liberati, N. T. et al. An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants. Proc. Natl Acad. Sci. 103, 2833–2838 (2006). · DOI · Paper page
  56. Hoang, T. T., Karkhoff-Schweizer, R. R., Kutchma, A. J. & Schweizer, H. P. A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 212, 77–86 (1998). · DOI · Paper page
  57. Jeske, A., Arce-Rodriguez, A., Thöming, J. G., Tomasch, J. & Häussler, S. Evolution of biofilm-adapted gene expression profiles in lasR-deficient clinical Pseudomonas aeruginosa isolates. NPJ Biofilms Microbiomes 8, 6 (2022). · DOI
  58. Wirth, N. T., Kozaeva, E. & Nikel, P. I. Accelerated genome engineering of Pseudomonas putida by I-SceI-mediated recombination and CRISPR-Cas9 counterselection. Microb. Biotechnol. 13, 233–249 (2020). · DOI
  59. Hodgkinson, J. T. et al. Robust routes for the synthesis of N-acylated-l-homoserine lactone (AHL) quorum sensing molecules with high levels of enantiomeric purity. Tetrahedron Lett. 52, 3291–3294 (2011). · doi:10.1016/j.tetlet.2011.04.059
  60. Studier, F. W. Protein production by auto-induction in high density shaking cultures. Protein Expr. Purif. 41, 207–234 (2005). · DOI · Paper page
  61. Hughes, C. S. et al. Single-pot, solid-phase-enhanced sample preparation for proteomics experiments. Nat. Protoc. 14, 68–85 (2019). · DOI · Paper page
  62. Erdmann, J., Preusse, M., Khaledi, A., Pich, A. & Häussler, S. Environment-driven changes of mRNA and protein levels in Pseudomonas aeruginosa. Environ. Microbiol. 20, 3952–3963 (2018). · DOI
  63. Cox, J. & Mann, M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat. Biotechnol. 26, 1367–1372 (2008). · DOI · Paper page
  64. Kabsch, W. XDS. Acta Crystallogr. D. Biol. Crystallogr. 66, 125–132 (2010). · DOI · Paper page
  65. Tickle, I. et al. STARANISO.Global Phasing Ltd., Cambridge, United Kingdom. http://staraniso.globalphasing.org/cgi-bin/staraniso.cgi. (2018).
  66. Winn, M. D. et al. Overview of the CCP4 suite and current developments. Acta Crystallogr. D. Biol. Crystallogr. 67, 235–242 (2011). · DOI · Paper page
  67. McCoy, A. J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007). · DOI · Paper page
  68. Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D. Biol. Crystallogr. 66, 486–501 (2010). · DOI · Paper page
  69. Liebschner, D. et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr. Sect. Struct. Biol. 75, 861–877 (2019). · DOI · Paper page
  70. Perez-Riverol, Y. et al. The PRIDE database resources in 2022: a hub for mass spectrometry-based proteomics evidences. Nucl. Acids Res. 50, D543–D552 (2022). · DOI · Paper page

Cited by

See all 93

Metadata

Authors
Borgert, S. R., Henke, S., Witzgall, F., Schmelz, S., zur Lage, S., Hotop, S.-K., Stephen, S., Lübken, D., Krüger, J., Gomez, N. O., van Ham, M., Jänsch, L., Kalesse, M., Pich, A., Brönstrup, M., Häussler, S., & Blankenfeldt, W.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2022-12-01
Volume
13
Issue
1
ISSN
2041-1723
DOI
10.1038/s41467-022-35030-w
Cited by
93
References
3
Language
en
Licence

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: Moonlighting chaperone activity of the enzyme PqsE contributes to RhlR-controlled virulence of Pseudomonas aeruginosa. https://app.cochat.ai/papers/10.1038/s41467-022-35030-w
The paper
Borgert, S. R., Henke, S., Witzgall, F., Schmelz, S., zur Lage, S., Hotop, S.-K., Stephen, S., Lübken, D., Krüger, J., Gomez, N. O., van Ham, M., Jänsch, L., Kalesse, M., Pich, A., Brönstrup, M., Häussler, S., & Blankenfeldt, W. (2022). Moonlighting chaperone activity of the enzyme PqsE contributes to RhlR-controlled virulence of Pseudomonas aeruginosa. Nature Communications. https://doi.org/10.1038/s41467-022-35030-w

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.