Force-Induced Changes of PilY1 Drive Surface Sensing by Pseudomonas aeruginosa
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Curated by eLife
Evaluation Summary:
This study demonstrates that a cysteine residue (C152) in the vWA domain of the type-iV pili tip-associated protein, PilY1 impacts surface sensing, biofilm formation and cyclic-di-GMP signaling in Pseduomonas aeruginosa. Well-executed experiments provide insight into the events that initiate cell adhesion and colonisation, the understanding of which has important implications for human health. The work will be of interest to microbiologists in general.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)
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Abstract
Most bacteria live on abiotic and biotic surfaces in surface-attached communities known as biofilms. Surface sensing and increased levels of the second-messenger molecule c-di-GMP are crucial to the transition from planktonic to biofilm growth.
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Evaluation Summary:
This study demonstrates that a cysteine residue (C152) in the vWA domain of the type-iV pili tip-associated protein, PilY1 impacts surface sensing, biofilm formation and cyclic-di-GMP signaling in Pseduomonas aeruginosa. Well-executed experiments provide insight into the events that initiate cell adhesion and colonisation, the understanding of which has important implications for human health. The work will be of interest to microbiologists in general.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #1 agreed to share their name with the authors.)
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Reviewer #1 (Public Review):
The authors wanted to evaluate the role of the vWA domain of the PilY1 protein in surface sensing in Pseudomonas aeruginosa. They specifically wanted to determine whether force induced conformational changes were necessary for adequate adhesion and the role this played in biofilm formation and cdG signaling. The authors very nicely used a combination of functional genetics, microbiological classical methods, comparative genomics and microscopy to evaluate the role of said domain, and more precisely that of cysteine-152 within vWA. They generally found that:
- There is a strong dependency on cysteine-152 (C152S), as its removal results in reduced biofilm formation and lower c-d-GMP levels, similarly as does the removal of the whole or partial vWA domain.
- C152S leads to a reduction in adhesion strength and a …
Reviewer #1 (Public Review):
The authors wanted to evaluate the role of the vWA domain of the PilY1 protein in surface sensing in Pseudomonas aeruginosa. They specifically wanted to determine whether force induced conformational changes were necessary for adequate adhesion and the role this played in biofilm formation and cdG signaling. The authors very nicely used a combination of functional genetics, microbiological classical methods, comparative genomics and microscopy to evaluate the role of said domain, and more precisely that of cysteine-152 within vWA. They generally found that:
- There is a strong dependency on cysteine-152 (C152S), as its removal results in reduced biofilm formation and lower c-d-GMP levels, similarly as does the removal of the whole or partial vWA domain.
- C152S leads to a reduction in adhesion strength and a significant change in the magnitude and frequency of spikes and plateaus, thereby altering their mechanical behaviors during surface adherence.
- Comparative genomic analysis of different strains of P. aeruginosa indicated that the cysteine residues in PA14 are highly conserved within that clade, but not in that of PAO1 and others. This in turn resulted in a functional deficiency to form biofilms relative to wtPA14.
- The results obtained in this study, and previous data obtained by the same team and others, led the authors to propose a model for force induced biofilm formation and transition from planktonic to irreversible attachment to surfaces, highlighting the potential interaction between PliY1 and PilO and the downstream signaling cascade it unfolds through the subsequent interaction between PilO and SadC and, ultimately, its effect on cdG.
Overall, the authors very systematically approached their hypotheses using appropriate methods. I think their conclusions are well supported by their data, their previous findings and that of other groups.
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Reviewer #2 (Public Review):
An interesting work investigating the role of Cysteine residues and disulfide bonds in protein mechanical stability and bacterial adhesion utilizing a variety of techniques to study the mechanical properties of the vWA associated PilY1 protein. The authors first study the ability of the bacterium to form biofilms. The authors investigate how PilY1 regulates c-di-GMP levels for biofilm formation and thereafter, they use AFM to prove surface adhesion forces as well as the effect of a Cys mutation (152S) in such adhesion. The work may have an impact in the field of mechanobiology provided that the authors complete their studies with experiments that involve the proteins alone. The work may be of general interest given the importance of the topic.
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