Exploring the transcriptome of luxI- and ΔainS mutants and the impact of N-3-oxo-hexanoyl-L- and N-3-hydroxy-decanoyl-L-homoserine lactones on biofilm formation in Aliivibrio salmonicida

Miriam Khider; Hilde Hansen; Erik Hjerde; Jostein A Johansen; Nils Peder Willassen

doi:10.7717/peerj.6845

Exploring the transcriptome of luxI^- and ΔainS mutants and the impact of N-3-oxo-hexanoyl-L- and N-3-hydroxy-decanoyl-L-homoserine lactones on biofilm formation in Aliivibrio salmonicida

PeerJ. 2019 Apr 30:7:e6845. doi: 10.7717/peerj.6845. eCollection 2019.

Authors

Miriam Khider¹, Hilde Hansen¹, Erik Hjerde^{1

2}, Jostein A Johansen¹, Nils Peder Willassen^{1

2}

Affiliations

¹ Norwegian Structural Biology Centre, Department of Chemistry, Faculty of Science and Technology, UiT-The Arctic University of Norway, Tromsø, Norway.
² Centre for Bioinformatics, Department of Chemistry, Faculty of Science and Technology, UiT-The Arctic University of Norway, Tromsø, Norway.

Abstract

Background: Bacterial communication through quorum sensing (QS) systems has been reported to be important in coordinating several traits such as biofilm formation. In Aliivibrio salmonicida two QS systems the LuxI/R and AinS/R, have been shown to be responsible for the production of eight acyl-homoserine lactones (AHLs) in a cell density dependent manner. We have previously demonstrated that inactivation of LitR, the master regulator of the QS system resulted in biofilm formation, similar to the biofilm formed by the AHL deficient mutant ΔainSluxI^- . In this study, we aimed to investigate the global gene expression patterns of luxI and ainS autoinducer synthases mutants using transcriptomic profiling. In addition, we examined the influence of the different AHLs on biofilm formation.

Results: The transcriptome profiling of ΔainS and luxI^- mutants allowed us to identify genes and gene clusters regulated by QS in A. salmonicida. Relative to the wild type, the ΔainS and luxI^- mutants revealed 29 and 500 differentially expressed genes (DEGs), respectively. The functional analysis demonstrated that the most pronounced DEGs were involved in bacterial motility and chemotaxis, exopolysaccharide production, and surface structures related to adhesion. Inactivation of luxI, but not ainS genes resulted in wrinkled colony morphology. While inactivation of both genes (ΔainSluxI^- ) resulted in strains able to form wrinkled colonies and mushroom structured biofilm. Moreover, when the ΔainSluxI^- mutant was supplemented with N-3-oxo-hexanoyl-L-homoserine lactone (3OC6-HSL) or N-3-hydroxy-decanoyl-L-homoserine lactone (3OHC10-HSL), the biofilm did not develop. We also show that LuxI is needed for motility and for repression of EPS production, where repression of EPS is likely operated through the RpoQ-sigma factor.

Conclusion: These findings imply that the LuxI and AinS autoinducer synthases play a critical role in the regulation of biofilm formation, EPS production, and motility.

Keywords: Acyl homoserine lactone; Aliivibrio salmonicida; Biofilm; Exopolysccharides; Quorum sensing.

Grants and funding

This work was funded by UiT, The Arctic University of Norway. The publication charges for this article have been funded by a grant from the publication fund of UiT, The Arctic University of Norway. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.